WO2011156836A1 - Apparatus for loading oligonucleotide spotting devices and spotting oligonucleotide probes - Google Patents
Apparatus for loading oligonucleotide spotting devices and spotting oligonucleotide probes Download PDFInfo
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- WO2011156836A1 WO2011156836A1 PCT/AU2011/000659 AU2011000659W WO2011156836A1 WO 2011156836 A1 WO2011156836 A1 WO 2011156836A1 AU 2011000659 W AU2011000659 W AU 2011000659W WO 2011156836 A1 WO2011156836 A1 WO 2011156836A1
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Definitions
- the present invention relates to diagnostic devices that use microsystems technologies (MST).
- MST microsystems technologies
- the invention relates to microfluidic and biochemical processing and analysis for molecular diagnostics.
- molecular diagnostic tests have the potential to reduce the occurrence of ineffective health care services, enhance patient outcomes, improve disease management and individualize patient care.
- Many of the techniques in molecular diagnostics are based on the detection and identification of specific nucleic acids, both deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), extracted and amplified from a biological specimen (such as blood or saliva).
- DNA deoxyribonucleic acid
- RNA ribonucleic acid
- the complementary nature of the nucleic acid bases allows short sequences of synthesized DNA (oligonucleotides) to bond (hybridize) to specific nucleic acid sequences for use in nucleic acid tests. If hybridization occurs, then the complementary sequence is present in the sample. This makes it possible, for example, to predict the disease a person will contract in the future, determine the identity and virulence of an infectious pathogen, or determine the response a person will have to a drug.
- a nucleic acid based test has four distinct steps:
- sample types are used for genetic analysis, such as blood, urine, sputum and tissue samples.
- the diagnostic test determines the type of sample required as not all samples are representative of the disease process. These samples have a variety of constituents, but usually only one of these is of interest. For example, in blood, high concentrations of erythrocytes can inhibit the detection of a pathogenic organism.
- Blood is one of the more commonly sought sample types. It has three major constituents: leukocytes (white blood cells), erythrocytes (red blood cells) and
- thrombocytes platelets.
- the thrombocytes facilitate clotting and remain active in vitro.
- the specimen is mixed with an agent such as
- EDTA ethylenediaminetetraacetic acid
- Erythrocytes are usually removed from the sample in order to concentrate the target cells. In humans, erythrocytes account for approximately 99% of the cellular material but do not carry DNA as they have no nucleus. Furthermore, erythrocytes contain components such as haemoglobin that can interfere with the downstream nucleic acid amplification process (described below). Removal of erythrocytes can be achieved by differentially lysing the erythrocytes in a lysis solution, leaving remaining cellular material intact which can then be separated from the sample using centrifugation. This provides a concentration of the target cells from which the nucleic acids are extracted.
- extracting nucleic acids from target cells usually involves a cell lysis step followed by nucleic acid purification.
- the cell lysis step disrupts the cell and nuclear membranes, releasing the genetic material. This is often accomplished using a lysis detergent, such as sodium dodecyl sulfate, which also denatures the large amount of proteins present in the cells.
- the nucleic acids are then purified with an alcohol precipitation step, usually ice- cold ethanol or isopropanol, or via a solid phase purification step, typically on a silica matrix in a column, resin or on paramagnetic beads in the presence of high concentrations
- GCA003-PCT of a chaotropic salt prior to washing and then elution in a low ionic strength buffer.
- An optional step prior to nucleic acid precipitation is the addition of a protease which digests the proteins in order to further purify the sample.
- lysis methods include mechanical lysis via ultrasonic vibration and thermal lysis where the sample is heated to 94°C to disrupt cell membranes.
- the target DNA or RNA may be present in the extracted material in very small amounts, particularly if the target is of pathogenic origin. Nucleic acid amplification provides the ability to selectively amplify (that is, replicate) specific targets present in low concentrations to detectable levels.
- PC polymerase chain reaction
- PCR is a powerful technique that amplifies a target DNA sequence against a background of complex DNA. If RNA is to be amplified (by PCR), it must be first transcribed into cDNA (complementary DNA) using an enzyme called reverse transcriptase. Afterwards, the resulting cDNA is amplified by PCR.
- PCR is an exponential process that proceeds as long as the conditions for sustaining the reaction are acceptable.
- the components of the reaction are:
- pair of primers short single strands of DNA with around 10-30 nucleotides complementary to the regions flanking the target sequence
- DNA polymerase - a thermostable enzyme that synthesizes DNA
- deoxyribonucleoside triphosphates (dNTPs) - provide the nucleotides that are incorporated into the newly synthesized DNA strand
- PCR typically involves placing these reactants in a small tube (-10-50 microlitres) containing the extracted nucleic acids.
- the tube is placed in a thermal cycler; an instrument that subjects the reaction to a series of different temperatures for varying amounts of time.
- the standard protocol for each thermal cycle involves a denaturation phase, an annealing phase, and an extension phase.
- the extension phase is sometimes referred to as the primer extension phase.
- two-step thermal protocols can be employed, in which the annealing and extension phases are combined.
- the denaturation phase typically involves raising the temperature of the
- GCA003-PCT reaction to 90 - 95°C to denature the DNA strands; in the annealing phase, the temperature is lowered to ⁇ 50-60°C for the primers to anneal; and then in the extension phase the temperature is raised to the optimal DNA polymerase activity temperature of 60- 72°C for primer extension. This process is repeated cyclically around 20-40 times, the end result being the creation of millions of copies of the target sequence between the primers.
- Multiplex PCR uses multiple primer sets within a single PCR mixture to produce amplicons of varying sizes that are specific to different DNA sequences. By targeting multiple genes at once, additional information may be gained from a single test-run that otherwise would require several experiments. Optimization of multiplex PCR is more difficult though and requires selecting primers with similar annealing temperatures, and amplicons with similar lengths and base composition to ensure the amplification efficiency of each amplicon is equivalent.
- Linker-primed PCR also known as ligation adaptor PCR
- ligation adaptor PCR is a method used to enable nucleic acid amplification of essentially all DNA sequences in a complex DNA mixture without the need for target-specific primers.
- the method firstly involves digesting the target DNA population with a suitable restriction endonuclease (enzyme). Double- stranded oligonucleotide linkers (also called adaptors) with a suitable overhanging end are then ligated to the ends of target DNA fragments using a ligase enzyme. Nucleic acid amplification is subsequently performed using oligonucleotide primers which are specific for the linker sequences. In this way, all fragments of the DNA source which are flanked by linker oligonucleotides can be amplified.
- enzyme double- stranded oligonucleotide linkers
- oligonucleotide primers which are specific for the linker sequences
- Direct PCR describes a system whereby PCR is performed directly on a sample without any, or with minimal, nucleic acid extraction. It has long been accepted that PCR reactions are inhibited by the presence of many components of unpurified biological samples, such as the haem component in blood. Traditionally, PCR has required extensive purification of the target nucleic acid prior to preparation of the reaction mixture. With appropriate changes to the chemistry and sample concentration, however, it is possible to perform PCR with minimal DNA purification, or direct PCR. Adjustments to the PCR chemistry for direct PCR include increased buffer strength, the use of polymerases which have high activity and processivity, and additives which chelate with potential polymerase inhibitors.
- Tandem PCR utilises two distinct rounds of nucleic acid amplification to increase the probability that the correct amplicon is amplified.
- One form of tandem PCR is nested PCR in which two pairs of PCR primers are used to amplify a single locus in separate rounds of nucleic acid amplification. The first pair of primers hybridize to the nucleic acid sequence at regions external to the target nucleic acid sequence. The second pair of primers (nested primers) used in the second round of amplification bind within the first PCR product and produce a second PCR product containing the target nucleic acid, that will be shorter than the first one.
- Real-time PCR or quantitative PCR, is used to measure the quantity of a PCR product in real time.
- a fluorophore-containing probe or fluorescent dyes along with a set of standards in the reaction, it is possible to quantitate the starting amount of nucleic acid in the sample. This is particularly useful in molecular diagnostics where treatment options may differ depending on the pathogen load in the sample.
- Reverse-transcriptase PCR is used to amplify DNA from RNA.
- Reverse transcriptase is an enzyme that reverse transcribes RNA into complementary DNA (cDNA), which is then amplified by PCR.
- cDNA complementary DNA
- RT-PCR is widely used in expression profiling, to determine the expression of a gene or to identify the sequence of an RNA transcript, including transcription start and termination sites. It is also used to amplify RNA viruses such as human immunodeficiency virus or hepatitis C virus.
- Isothermal amplification is another form of nucleic acid amplification which does not rely on the thermal denaturation of the target DNA during the amplification reaction and hence does not require sophisticated machinery. Isothermal nucleic acid amplification methods can therefore be carried out in primitive sites or operated easily outside of a laboratory environment. A number of isothermal nucleic acid amplification methods have been described, including Strand Displacement Amplification, Transcription Mediated Amplification, Nucleic Acid Sequence Based Amplification, Recombinase Polymerase Amplification, Rolling Circle Amplification, Ramification Amplification, Helicase-
- Isothermal nucleic acid amplification methods do not rely on the continuing heat denaturation of the template DNA to produce single stranded molecules to serve as templates for further amplification, but instead rely on alternative methods such as
- GCA003-PCT enzymatic nicking of DNA molecules by specific restriction endonucleases, or the use of an enzyme to separate the DNA strands, at a constant temperature.
- Strand Displacement Amplification relies on the ability of certain restriction enzymes to nick the unmodified strand of hemi-modified DNA and the ability of a 5 '-3' exonuclease-deficient polymerase to extend and displace the downstream strand.
- Exponential nucleic acid amplification is then achieved by coupling sense and antisense reactions in which strand displacement from the sense reaction serves as a template for the antisense reaction.
- nickase enzymes which do not cut DNA in the traditional manner but produce a nick on one of the DNA strands, such as N. Alwl, N. BstNBl and Mly 1, are useful in this reaction.
- SDA has been improved by the use of a combination of a heat-stable restriction enzyme (Aval) and heat-stable Exo- polymerase (Bst polymerase). This combination has been shown to increase amplification efficiency of the reaction from 10 8 fold amplification to 10 10 fold amplification so that it is possible using this technique to amplify unique single copy molecules.
- TMA Transcription Mediated Amplification
- RNA polymerase uses an RNA polymerase to copy RNA sequences but not corresponding genomic DNA.
- the technology uses two primers and two or three enzymes, RNA polymerase, reverse transcriptase and optionally RNase H (if the reverse transcriptase does not have RNase activity).
- One primer contains a promoter sequence for RNA polymerase.
- this primer hybridizes to the target ribosomal RNA (rRNA) at a defined site.
- rRNA ribosomal RNA
- Reverse transcriptase creates a DNA copy of the target rRNA by extension from the 3' end of the promoter primer.
- RNA in the resulting RNA:DNA duplex is degraded by the RNase activity of the reverse transcriptase if present or the additional RNase H.
- a second primer binds to the DNA copy.
- a new strand of DNA is synthesized from the end of this primer by reverse transcriptase, creating a double-stranded DNA molecule.
- RNA polymerase recognizes the promoter sequence in the DNA template and initiates transcription. Each of the newly synthesized RNA amplicons re-enters the process and serves as a template for a new round of replication.
- RPA Recombinase Polymerase Amplification
- GCA003-PCT structures are stabilised by single-stranded DNA binding proteins interacting with the displaced template strand, thus preventing the ejection of the primer by branch migration.
- Recombinase disassembly leaves the 3' end of the oligonucleotide accessible to a strand displacing DNA polymerase, such as the large fragment of Bacillus subtilis Pol I (Bsu), and primer extension ensues. Exponential nucleic acid amplification is accomplished by the cyclic repetition of this process.
- HSA Helicase-dependent amplification
- a DNA helicase enzyme to generate single-stranded templates for primer hybridization and subsequent primer extension by a DNA polymerase.
- the helicase enzyme traverses along the target DNA, disrupting the hydrogen bonds linking the two strands which are then bound by single-stranded binding proteins. Exposure of the single-stranded target region by the helicase allows primers to anneal.
- the DNA polymerase then extends the 3 ' ends of each primer using free deoxyribonucleoside triphosphates (dNTPs) to produce two DNA replicates.
- dNTPs free deoxyribonucleoside triphosphates
- RCA Rolling Circle Amplification
- a DNA polymerase extends a primer continuously around a circular DNA template, generating a long DNA product that consists of many repeated copies of the circle.
- the polymerase generates many thousands of copies of the circular template, with the chain of copies tethered to the original target DNA.
- Ramification amplification is a variation of RCA and utilizes a closed circular probe (C-probe) or padlock probe and a DNA polymerase with a high processivity to exponentially amplify the C-probe under isothermal conditions.
- Loop-mediated isothermal amplification offers high selectivity and employs a DNA polymerase and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA.
- An inner primer containing sequences of the sense and antisense strands of the target DNA initiates LAMP.
- the following strand displacement DNA synthesis primed by an outer primer releases a single-stranded DNA. This serves as template for DNA synthesis primed by the second inner and outer primers that hybridize to the other end of the target, which produces a stem-loop DNA structure.
- one inner primer hybridizes to the loop on the product and
- GCA003-PCT initiates displacement DNA synthesis, yielding the original stem-loop DNA and a new stem-loop DNA with a stem twice as long.
- the cycling reaction continues with accumulation of 10 9 copies of target in less than an hour.
- the final products are stem-loop DNAs with several inverted repeats of the target and cauliflower-like structures with multiple loops formed by annealing between alternately inverted repeats of the target in the same strand.
- the amplified product After completion of the nucleic acid amplification, the amplified product must be analysed to determine whether the anticipated amplicon (the amplified quantity of target nucleic acids) was generated.
- the methods of analyzing the product range from simply determining the size of the amplicon through gel electrophoresis, to identifying the nucleotide composition of the amplicon using DNA hybridization.
- Gel electrophoresis is one of the simplest ways to check whether the nucleic acid amplification process generated the anticipated amplicon.
- Gel electrophoresis uses an electric field applied to a gel matrix to separate DNA fragments. The negatively charged DNA fragments will move through the matrix at different rates, determined largely by their size. After the electrophoresis is complete, the fragments in the gel can be stained to make them visible. Ethidium bromide is a commonly used stain which fluoresces under UV light.
- the size of the fragments is determined by comparison with a DNA size marker (a DNA ladder), which contains DNA fragments of known sizes, run on the gel alongside the amplicon. Because the oligonucleotide primers bind to specific sites flanking the target DNA, the size of the amplified product can be anticipated and detected as a band of known size on the gel. To be certain of the identity of the amplicon, or if several amplicons have been generated, DNA probe hybridization to the amplicon is commonly employed.
- a DNA size marker a DNA ladder
- DNA hybridization refers to the formation of double-stranded DNA by
- DNA hybridization for positive identification of a specific amplification product requires the use of a DNA probe around 20 nucleotides in length. If the probe has a sequence that is complementary to the amplicon (target) DNA sequence, hybridization will occur under favourable conditions of temperature, pH and ionic concentration. If hybridization occurs, then the gene or DNA sequence of interest was present in the original sample.
- Optical detection is the most common method to detect hybridization. Either the amplicons or the probes are labelled to emit light through fluorescence or
- GCA003-PCT of the light-producing moieties enable covalent labelling of nucleotide strands.
- ECL electrochemiluminescence
- fluorescence it is illumination with excitation light which leads to emission.
- Fluorescence is detected using an illumination source which provides excitation light at a wavelength absorbed by the fluorescent molecule, and a detection unit.
- the detection unit comprises a photosensor (such as a photomultiplier tube or charge-coupled device (CCD) array) to detect the emitted signal, and a mechanism (such as a wavelength- selective filter) to prevent the excitation light from being included in the photosensor output.
- the fluorescent molecules emit Stokes -shifted light in response to the excitation light, and this emitted light is collected by the detection unit. Stokes shift is the frequency difference or wavelength difference between emitted light and absorbed excitation light.
- ECL emission is detected using a photosensor which is sensitive to the emission wavelength of the ECL species being employed.
- a photosensor which is sensitive to the emission wavelength of the ECL species being employed.
- transition metal-ligand complexes emit light at visible wavelengths, so conventional photodiodes and CCDs are employed as photosensors.
- An advantage of ECL is that, if ambient light is excluded, the ECL emission can be the only light present in the detection system, which improves sensitivity.
- Microarrays allow for hundreds of thousands of DNA hybridization experiments to be performed simultaneously. Microarrays are powerful tools for molecular diagnostics with the potential to screen for thousands of genetic diseases or detect the presence of numerous infectious pathogens in a single test.
- a microarray consists of many different DNA probes immobilized as spots on a substrate. The target DNA (amplicon) is first labelled with a fluorescent or luminescent molecule (either during or after nucleic acid amplification) and then applied to the array of probes. The microarray is incubated in a temperature controlled, humid environment for a number of hours or days while hybridization between the probe and amplicon takes place. Following incubation, the microarray must be washed in a series of buffers to remove unbound strands.
- the microarray surface is dried using a stream of air (often nitrogen).
- the stringency of the hybridization and washes is critical. Insufficient stringency can result in a high degree of nonspecific binding. Excessive stringency can lead to a failure of appropriate binding, which results in diminished sensitivity.
- Hybridization is recognized by detecting light emission from the labelled amplicons which have formed a hybrid with complementary probes.
- GCA003-PCT Fluorescence from microarrays is detected using a microarray scanner which is generally a computer controlled inverted scanning fluorescence confocal microscope which typically uses a laser for excitation of the fluorescent dye and a photosensor (such as a photomultiplier tube or CCD) to detect the emitted signal.
- the fluorescent molecules emit Stokes-shifted light (described above) which is collected by the detection unit.
- the emitted fluorescence must be collected, separated from the unabsorbed excitation wavelength, and transported to the detector.
- a confocal arrangement is commonly used to eliminate out-of- focus information by means of a confocal pinhole situated at an image plane. This allows only the in- focus portion of the light to be detected. Light from above and below the plane of focus of the object is prevented from entering the detector, thereby increasing the signal to noise ratio.
- the detected fluorescent photons are converted into electrical energy by the detector which is subsequently converted to a digital signal. This digital signal translates to a number representing the intensity of fluorescence from a given pixel. Each feature of the array is made up of one or more such pixels.
- the final result of a scan is an image of the array surface. The exact sequence and position of every probe on the microarray is known, and so the hybridized target sequences can be identified and analysed simultaneously.
- a point-of-care technology serving the physician's office, the hospital bedside or even consumer-based, at home, would offer many advantages including:
- GCA003-PCT • rapid availability of results enabling immediate facilitation of treatment and improved quality of care.
- LOC Lab-on-a-chip
- microfluidic device for analyzing a sample fluid, the microfluidic device comprising:
- MST microsystems technology
- CMOS circuitry with digital memory for storing data and operational information to operatively control the functional sections during processing and analysis of the sample.
- the microfluidic device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory relates to the reagent identities.
- the data stored in the digital memory is a unique identifier for the microfluidic device.
- the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
- the microfluidic device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
- the microfluidic device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
- each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and
- the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
- the data stored in the digital memory includes hybridization data generated from the photosensor output.
- the micro fluidic device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the CMOS circuitry has bond-pads and is configured for transmission of the hybridization data to an external device.
- the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the bond-pads and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
- the micro fluidic device also has a supporting substrate and a cap wherein the CMOS circuitry is between the supporting substrate and the MST layer, and the cap overlies the MST layer and defines the reagent reservoirs.
- the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
- the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
- the easily usable, mass-producible, and inexpensive microfluidic device with integral digital memory accepts an input fluid and processes it.
- the digital memory is used to store the data and control information required during the functioning of the device and the module incorporating the device.
- the digital memory being integral to the device, provides for an easily manufacturable, mass-producible, easily usable, and inexpensive microfluidic system with low component-count.
- test module for analyzing a sample fluid, the test module comprising:
- a receptacle for receiving the sample
- digital memory for storing data and operational information to operatively control the functional sections during processing and analysis of the sample.
- the test module also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory relates to the reagent identities.
- the data stored in the digital memory is a unique identifier for the test module.
- the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
- the test module also has CMOS circuitry and a temperature sensor wherein the CMOS circuitry incorporates the digital memory and uses the temperature sensor output for feedback control of the incubation section.
- the test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
- each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
- the data stored in the digital memory includes hybridization data generated from the photosensor output.
- the test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the CMOS circuitry has a data interface for transmission of the hybridization data to an external device.
- the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the data interface and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the control circuitry.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
- the test module also has a LOC device having a sample inlet in fluid communication with the receptacle, a supporting substrate, a microsystems technology (MST) layer, CMOS circuitry between the supporting substrate and the MST layer and a cap wherein the CMOS circuitry incorporates the digital memory, the MST layer incorporates the functional sections, and the cap overlies the MST layer and defines the reagent reservoirs.
- MST microsystems technology
- the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that
- GCA003-PCT contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
- the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
- the easily usable, mass-producible, and inexpensive LOC device with integral digital memory accepts an input fluid and processes it.
- the digital memory is used to store the data and control information required during the functioning of the LOC device and the module incorporating the LOC device.
- the information stored on the memory includes the characteristics of the module incorporating this LOC device.
- the digital memory being integral to the device, provides for an easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
- This aspect of the invention provides a lab-on-a-chip (LOC) device for analyzing a sample fluid, the LOC device comprising:
- MST microsystems technology
- CMOS circuitry with digital memory for storing epidemiological data, and configured to download epidemiological data updates from an external source.
- the CMOS circuitry incorporates a universal serial bus (USB) device driver for operative control of a USB connection to the external source.
- USB universal serial bus
- the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory relates to the reagent identities.
- the data stored in the digital memory is a unique identifier for LOC device.
- the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an enzymatic reaction temperature.
- the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
- the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
- the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the CMOS circuitry has bond-pads for connection to the USB connection and transmission of hybridization data to an external device.
- the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the USB connection and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
- the LOC device also has a supporting substrate and a cap wherein the CMOS circuitry is between the supporting substrate and the MST layer, and the cap overlies the MST layer and defines the reagent reservoirs.
- the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that
- GCA003-PCT contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
- the PCR section has a thermal cycle time of less than 4 seconds.
- the PCR section has a thermal cycle time between 0.45 seconds and 1.5 seconds.
- the easily usable, mass-producible, and inexpensive LOC device with integral digital memory accepts an input fluid and processes it.
- the digital memory is used to store the data and control information required during the functioning of the LOC device and the module incorporating the LOC device.
- the information stored in the memory includes epidemiological updates available at the time, with the information being used for analytical and diagnostics purposes. This information provides for module's independence from outside support.
- the digital memory being integral to the device, provides for an easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
- This aspect of the invention provides a lab-on-a-chip (LOC) device for analyzing a sample fluid, the LOC device comprising:
- MST microsystems technology
- CMOS circuitry with digital memory for storing genetic data, and configured to download genetic data updates from an external source.
- the CMOS circuitry incorporates a universal serial bus
- USB universal serial Bus
- the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory relates to the reagent identities.
- the data stored in the digital memory is a unique identifier for LOC device.
- the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the genetic material.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an enzymatic reaction temperature.
- the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
- the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
- the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the CMOS circuitry has bond-pads for transmission of hybridization data via the USB connection.
- the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the bond-pads and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
- the LOC device also has a supporting substrate and a cap wherein the CMOS circuitry is between the supporting substrate and the MST layer, and the cap overlies the MST layer and defines the reagent reservoirs.
- the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
- the PCR section has a thermal cycle time of less than 4 seconds.
- the PCR section has a thermal cycle time between 0.45 seconds and 1.5 seconds.
- the easily usable, mass-producible, and inexpensive LOC device with integral digital memory accepts an input fluid and processes it.
- the digital memory is used to store the data and control information required during the functioning of the LOC device and the module incorporating the LOC device.
- the information stored in the memory includes genetic information updates available at the time, with the information being used for analytical and diagnostics purposes. This information provides for module's independence from outside support.
- the digital memory being integral to the device, provides for an easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
- This aspect of the invention provides a lab-on-a-chip (LOC) device for analyzing a sample fluid, the LOC device comprising:
- MST microsystems technology
- CMOS circuitry with digital memory for storing data and operational information to operatively control the functional sections during processing and analysis of the sample;
- the data is encrypted for secure communication with an external device.
- the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory relates to the reagent identities.
- the data stored in the digital memory is a unique identifier for LOC device.
- the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
- the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
- the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
- each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
- the data stored in the digital memory includes hybridization data generated from the photosensor output.
- the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the CMOS circuitry has bond-pads and is configured for transmission of the hybridization data to an external device.
- the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the bond-pads and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
- the LOC device also has a supporting substrate and a cap wherein the CMOS circuitry is between the supporting substrate and the MST layer, and the cap overlies the MST layer and defines the reagent reservoirs.
- the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
- the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
- the easily usable, mass-producible, and inexpensive LOC device with an integral digital memory accepts a diagnostic sample and processes it.
- the digital memory is used to store the data and control information required during the functioning of the LOC device and the diagnostic module incorporating the LOC device.
- the memory also securely stores patient test result information.
- the capability to store patient test result information makes it possible for the diagnostic module to perform a test utilizing only a minimal external power supply, and then in conjunction with a fully featured reader, analyze the patient test results at a later time.
- the secure storage of patient test result information would insure that the information would not be misused through illicit channels.
- the digital memory being integral to the device, provides for an easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
- test module for analyzing a sample fluid, the test module comprising:
- a receptacle for receiving the sample
- CMOS circuitry on the supporting substrate for operative control of the functional sections during processing and analysis of the sample.
- the test module also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the CMOS circuitry has digital memory for storing data relating to the reagent identities.
- the data stored in the digital memory is a unique identifier for test module.
- the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
- the test module also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
- the test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
- each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
- the data stored in the digital memory includes hybridization data generated from the photosensor output.
- the test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- control circuitry has a data interface for transmission of the hybridization data to an external device.
- the sample is drawn from a patient and the control circuitry is configured to download patient data via the data interface and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the control circuitry.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
- the test module also has a LOC device that incorporates the supporting substrate and the CMOS circuitry, and has a sample inlet in fluid communication with the receptacle, a microsystems technology (MST) layer that incorporates the functional sections and a cap wherein the cap overlies the MST layer and defines the reagent reservoirs.
- MST microsystems technology
- the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
- the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
- the easily usable, mass-producible, and inexpensive LOC device accepts a diagnostic sample and processes it, with the LOC device's on-chip electronics controlling all of the LOC device's functions.
- the control electronics being integral to the LOC device, makes the module utilizing the LOC device independent from specialized outside support and provides for the easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
- This aspect of the invention provides a lab-on-a-chip (LOC) device for installation in a test module for analyzing a sample fluid and communicating test results to an external device, the LOC device comprising:
- MST microsystems technology
- CMOS circuitry on the supporting substrate for operative control of a
- the CMOS circuitry incorporates a universal serial bus (USB) device driver for operative control of a USB plug in the test module for communication with an external device.
- USB universal serial bus
- the CMOS circuitry is further configured for operative control of the functional sections during processing and analysis of the sample.
- the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the CMOS circuitry has digital memory for storing data relating to the reagent identities.
- the data stored in the digital memory is a unique identifier for LOC device.
- the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample
- GCA003-PCT and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
- the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
- the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
- each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
- the data stored in the digital memory includes hybridization data generated from the photosensor output.
- the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the CMOS circuitry is configured for transmission of the hybridization data to an external device via the test module communications interface.
- the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the communications interface and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the control circuitry.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
- the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
- the easily usable, mass-producible, and inexpensive LOC device accepts a diagnostic sample and processes it, with the LOC device's on-chip electronics controlling the data and command communications with the host.
- the electronics being integral to the LOC device, makes the module utilizing the LOC device independent from specialized outside support and provides for the easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
- This aspect of the invention provides a lab-on-a-chip (LOC) device for installation in a test module for analyzing a sample fluid and communicating test results to an external device, the LOC device comprising:
- MST microsystems technology
- CMOS circuitry on the supporting substrate, the CMOS circuitry having a universal serial bus (USB) device driver for operative control of a USB plug in the test module for communication with the external device.
- USB universal serial bus
- the CMOS circuitry is further configured for operative control of the functional sections during processing and analysis of the sample.
- the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the CMOS circuitry has digital memory for storing data relating to the reagent identities.
- the data stored in the digital memory is a unique identifier for LOC device.
- the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
- the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
- the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
- each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
- the data stored in the digital memory includes hybridization data generated from the photosensor output.
- the digital memory includes random access memory
- RAM random access memory
- flash memory volatile and flash memory
- the RAM being configured to store the hybridization data
- the flash memory being configured to store program data to operate the functional sections and the probe identity data.
- the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the USB device driver is configured for transmission of the hybridization data to an external device via the USB plug.
- the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the USB plug and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the control circuitry.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
- the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
- the easily usable, mass-producible, and inexpensive LOC device with integral USB device controller accepts a diagnostic sample and processes it.
- the USB device controller being integral to the LOC device, makes the module utilizing the LOC device independent from specialized outside support and provides for the easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
- This aspect of the invention provides a lab-on-a-chip (LOC) device for analyzing a sample fluid, the LOC device comprising:
- MST microsystems technology
- the CMOS circuitry between the supporting substrate and the MST layer, the CMOS circuitry having a controller to control operations performed by the functional sections during processing and analysis of the sample.
- the CMOS circuitry has digital memory for storing data and operational information for use by the controller to control the functional sections during processing and analysis of the sample.
- the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory is a unique identifier for LOC device, the unique identifier being associated with the reagent identities.
- the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
- the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
- the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
- each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
- the data stored in the digital memory includes hybridization data generated from the photosensor output.
- the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the CMOS circuitry has bond-pads and is configured for transmission of the hybridization data to an external device.
- the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the bond-pads and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
- the LOC device also has a cap wherein the cap overlies the MST layer and defines the reagent reservoirs.
- the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
- the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
- the easily usable, mass-producible, and inexpensive LOC device accepts a diagnostic sample and processes it, with the LOC device's integral controller controlling all of the LOC device's functions.
- the controller being integral to the LOC device, makes the module utilizing the LOC device independent from specialized outside support and provides for the easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
- This aspect of the invention provides a lab-on-a-chip (LOC) device for analyzing a sample fluid, the LOC device comprising:
- MST microsystems technology
- CMOS circuitry between the supporting substrate and the MST layer, the CMOS circuitry having digital memory for storing data and operational information to operatively control the functional sections during processing and analysis of the sample.
- the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory relates to the reagent identities.
- the data stored in the digital memory is a unique identifier for LOC device.
- the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
- the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
- the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
- each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
- the data stored in the digital memory includes hybridization data generated from the photosensor output.
- the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the CMOS circuitry has bond-pads and is configured for transmission of the hybridization data to an external device.
- the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the bond-pads and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
- the LOC device also has a cap wherein the cap overlies the MST layer and defines the reagent reservoirs.
- the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
- the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
- the easily usable, mass-producible, and inexpensive LOC device accepts a diagnostic sample and processes it, with the LOC device's integral data RAM providing for intermediate data storage.
- the data RAM being integral to the LOC device, makes the module utilizing the LOC device independent from specialized outside support and provides for the easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
- LOC lab-on-a-chip
- GCA003-PCT a microsystems technology (MST) layer with functional sections for processing and analyzing the sample; and,
- CMOS circuitry between the supporting substrate and the MST layer, the CMOS circuitry having flash memory for storing program data to operate the functional sections during processing and analysis of the sample.
- the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the flash memory also stores data relating to the reagent identities.
- the data includes a unique identifier for LOC device.
- the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
- the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
- the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- the flash memory stores probe identity data identifying the probe at each site within the array of probes.
- each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
- the CMOS circuitry has random access memory (RAM) configured to store hybridization data generated from the photosensor output.
- RAM random access memory
- the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the CMOS circuitry has bond-pads and is configured for transmission of the hybridization data to an external device.
- the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the bond-pads and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
- the LOC device also has a cap wherein the cap overlies the MST layer and defines the reagent reservoirs.
- the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
- the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
- the easily usable, mass-producible, and inexpensive microfluidic device with integral program and data flash memory accepts an input fluid and processes it.
- the flash memory is used to store the data and program required during the functioning of the device and the module incorporating the device.
- the flash memory being integral to the device, provides for an easily manufacturable, mass-producible, easily usable, and inexpensive microfluidic system with low component-count.
- test module for analyzing a sample fluid and communicating epidemiological data to a database, the test module comprising:
- a receptacle for receiving the sample
- a controller for operative control of the communication interface.
- the test module also has a universal serial bus (USB) plug wherein the communication interface is a device driver for operative control of the USB plug to communicate with an external device.
- USB universal serial bus
- the test module also has digital memory for storing epidemiological data.
- the test module also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory includes the reagent identities.
- the data stored in the digital memory includes a unique identifier for test module.
- the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an enzymatic reaction temperature.
- the test module also has CMOS circuitry and a temperature sensor wherein the CMOS circuitry incorporates the digital memory and uses the temperature sensor output for feedback control of the incubation section.
- the test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to excitation, and
- the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
- the test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the CMOS circuitry is configured for communication of hybridization data with an external device.
- the sample is drawn from a patient and the CMOS circuitry is configured to download and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
- the test module also has a LOC device having a sample inlet in fluid communication with the receptacle, a supporting substrate, a microsystems technology (MST) layer, CMOS circuitry between the supporting substrate and the MST layer and a cap wherein the CMOS circuitry incorporates the digital memory, the communication interface, the controller, the MST layer incorporates the functional sections, and the cap overlies the MST layer and defines the reagent reservoirs.
- MST microsystems technology
- the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
- the PCR section has a thermal cycle time of less than 4 seconds.
- the easily usable, mass-producible, inexpensive, and portable diagnostic test module accepts a biochemical sample and processes and analyzes the sample, updating epidemiological databases based on the diagnostic results derived from the sample.
- the updating of epidemiological databases provides for improved science-base for the functioning of the diagnostic test modules and optimal higher- level responses to epidemiological situations.
- the diagnostic test module based automation of updating the databases would provide for massive quality and economic gains for health information systems.
- test module for analyzing a sample fluid and communicating location data to an epidemiological database, the test module comprising:
- a receptacle for receiving the sample
- controller for operative control of the communication interface; wherein, the controller is configured to associate location data with epidemiological data sent to the communication interface for communication with the epidemiological database.
- the test module also has a universal serial bus (USB) plug wherein the communication interface is a USB device driver for operative control of the USB plug to communicate with an external device.
- USB universal serial bus
- the controller is configured to automatically
- the test module also has a user interface for inputting data to the controller for communication with the epidemiological database.
- the test module also has digital memory for storing epidemiological data.
- the test module also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory includes the reagent identities.
- the data stored in the digital memory includes a unique identifier for test module.
- the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample.
- PCR polymerase chain reaction
- the test module also has CMOS circuitry and a temperature sensor wherein the CMOS circuitry incorporates the digital memory and uses the temperature sensor output for feedback control of the PCR section.
- the test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to excitation, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
- the test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the CMOS circuitry is configured for communication of hybridization data to an external device.
- the sample is drawn from a patient and the CMOS circuitry is configured to download and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
- the test module also has a LOC device having a sample inlet in fluid communication with the receptacle, a supporting substrate, a microsystems technology (MST) layer, CMOS circuitry between the supporting substrate and the MST
- MST microsystems technology
- GCA003-PCT layer and a cap wherein the CMOS circuitry incorporates the digital memory, the communication interface, the controller, the MST layer incorporates the functional sections, and the cap overlies the MST layer and defines the reagent reservoirs.
- the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
- the easily usable, mass-producible, inexpensive, and portable diagnostic test module accepts a biochemical sample and processes and analyzes the sample, updating epidemiological databases based on the diagnostic results derived from the sample and the test location data.
- the updating of epidemiological databases with the diagnostics results and the location data provides for improved science-base for the functioning of the diagnostic test modules and optimal higher-level responses to epidemiological situations.
- the diagnostic test module based automation of updating the databases would provide for massive quality and economic gains for health information systems.
- test module for analyzing a sample fluid and communicating data to a medical database, the test module comprising: a receptacle for receiving the sample;
- a communication interface for communication with the medical database; and, a controller for operative control of the communication interface.
- test module of claim 1 further comprising a universal serial bus (USB) plug wherein the communication interface is a USB device driver for operative control of the USB plug to communicate with an external device.
- USB universal serial bus
- test module of claim 2 further comprising digital memory wherein the medical database stores electronic health records (EH ), electronic medical records (EMR) and personal health records (PHR) and, the digital memory is configured for storing data relating to EHR, EMR and PHR.
- EHR electronic health records
- EMR electronic medical records
- PHR personal health records
- test module of claim 3 further comprising a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory includes the reagent identities.
- the data stored in the digital memory includes a unique identifier for test module.
- the sample is a biological sample including cells of different sizes, and one of the functional sections is a polymerase chain reaction (PC ) section for amplifying nucleic acid sequences in the sample.
- PC polymerase chain reaction
- test module of claim 6 further comprising CMOS circuitry and a temperature sensor wherein the CMOS circuitry incorporates the digital memory and uses the temperature sensor output for feedback control of the PCR section.
- one of the functional sections is a dialysis section, the dialysis section being configured for separating cells larger than a predetermined threshold into a portion of the sample which is processed separately from the remainder of the sample containing only cells smaller than the predetermined threshold.
- one of the functional sections is a lysis section, the lysis section being configured to release nucleic acid sequences within the cells smaller than the predetermined threshold.
- test module of claim 9 further comprising an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to excitation, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
- test module of claim 1 1 further comprising a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the CMOS circuitry is configured to communicate hybridization data to an external device.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
- the test module of claim 5 further comprising a LOC device having a sample inlet in fluid communication with the receptacle, a supporting substrate, a microsystems technology (MST) layer, CMOS circuitry between the supporting substrate and the MST layer and a cap wherein the CMOS circuitry incorporates the digital memory, the communication interface, the controller, the MST layer incorporates the functional sections, and the cap overlies the MST layer and defines the reagent reservoirs.
- MST microsystems technology
- the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
- the PCR section has a thermal cycle time of less than 4 seconds.
- the easily usable, mass-producible, inexpensive, and portable diagnostic test module accepts a biochemical sample and processes and analyzes the sample, updating patients' databases based on the diagnostic results derived from the sample.
- the updating of patients' databases with the diagnostics results and the location data provides for improved provision of health care for the patients, automated maintenance of patient's medical records, improved science-base for the functioning of the diagnostic test modules, and optimal higher-level responses to epidemiological situations.
- the diagnostic test module based automation of updating the databases would provide for massive quality and economic gains for health information systems.
- GCA003-PCT GRE001.1 This aspect of the invention provides a micro flui die test module for analyzing a sample fluid and communicating test results to a mobile telephone, the microfluidic test module comprising:
- a communication interface for communication with the mobile telephone; and, a controller for operative control of the communication interface.
- the communication interface is a universal serial bus (USB) device driver for operative control of a USB plug in the test module for communication with an external device.
- USB universal serial bus
- the USB device driver is configured to draw power from the mobile telephone to power the controller and the functional sections.
- the controller is further configured for operative control of the functional sections during processing and analysis of the sample.
- the controller is configured to download data via the mobile telephone.
- the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing data relating to the reagent identities.
- the data stored in the digital memory is a unique identifier for the microfluidic test module.
- the sample is a biological sample containing genetic material and one of the functional sections is a nucleic acid amplification section for amplifying nucleic acid sequences in the genetic material.
- the nucleic acid amplification section is a polymerase chain reaction (PCR) section and the data stored in the digital memory includes thermal cycle times and cycle numbers.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
- the microfluidic test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
- the micro fluidic test module also has a photosensor wherein each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe- target hybrids being configured to emit photons in response to an input, and the photosensor is configured for sensing the photons emitted by the probe-target hybrids.
- the data stored in the digital memory includes hybridization data generated from the photosensor output.
- the micro fluidic test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the controller is configured for transmission of the hybridization data to the mobile telephone.
- the sample is drawn from a patient and the controller is configured to download patient data via the mobile telephone and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the controller.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the easily usable, mass-producible, inexpensive, compact, light, and portable Micro fluidic test module accepts a sample and processes and analyzes the sample material using the module's integral sensors, and provides the results electronically at its output port.
- the module's communications port is interfaced to a mobile phone/smart phone which provides the module with power, computing, communications, and user interface support.
- GCA003-PCT phones/smart phones are widely and inexpensively available, obviating the need for specialized, heavy, and expensive module support systems.
- This aspect of the invention provides a microfluidic test module for analyzing a sample fluid and communicating test results to a laptop computer, the microfluidic test module comprising:
- a communication interface for communication with the laptop computer; and, a controller for operative control of the communication interface.
- the communication interface is a universal serial bus (USB) device driver for operative control of a USB plug in the test module for communication with an external device.
- USB universal serial bus
- the USB device driver is configured to draw power from the laptop computer to power the controller and the functional sections.
- the controller is further configured for operative control of the functional sections during processing and analysis of the sample.
- the controller is configured to download data via the laptop computer.
- the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing data relating to the reagent identities.
- the data stored in the digital memory is a unique identifier for microfluidic test module.
- the sample is a biological sample containing genetic material and one of the functional sections is a nucleic acid amplification section for amplifying nucleic acid sequences in the genetic material.
- the nucleic acid amplification section is a polymerase chain reaction (PCR) section and the data stored in the digital memory includes thermal cycle times and cycle numbers.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample
- GCA003-PCT and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
- the microfluidic test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
- the microfluidic test module also has a photosensor wherein each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe- target hybrids being configured to emit photons in response to an input, and the photosensor is configured for sensing the photons emitted by the probe-target hybrids.
- the data stored in the digital memory includes hybridization data generated from the photosensor output.
- the microfluidic test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the controller is configured for transmission of the hybridization data to the laptop computer.
- the sample is drawn from a patient and the controller is configured to download patient data via the laptop computer and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the controller.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the easily usable, mass-producible, inexpensive, compact, light, and portable Microfluidic test module accepts a sample and processes and analyzes the sample material using the module's integral
- the module's communications port is interfaced to a laptop/notebook which provides the module with power, computing, communications, and user interface support.
- Laptops/notebooks are widely and inexpensively available, obviating the need for specialized, heavy, and expensive module support systems.
- This aspect of the invention provides a micro flui die test module for analyzing a sample fluid and communicating test results to a dedicated reader purpose built for operating with the microfluidic test module, the microfluidic test module comprising: an outer casing with a receptacle for receiving the sample;
- a communication interface for communication with the dedicated reader; and, a controller for operative control of the communication interface.
- the communication interface is a universal serial bus (USB) device driver for operative control of a USB plug in the test module for communication with an external device.
- USB universal serial bus
- the USB device driver is configured to draw power from the dedicated reader to power the controller and the functional sections.
- the controller is further configured for operative control of the functional sections during processing and analysis of the sample.
- the controller is configured to download data via the dedicated reader.
- the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing data relating to the reagent identities.
- the data stored in the digital memory is a unique identifier for microfluidic test module.
- the sample is a biological sample containing genetic material and one of the functional sections is a nucleic acid amplification section for amplifying nucleic acid sequences in the genetic material.
- the nucleic acid amplification section is a polymerase chain reaction (PCR) section and the data stored in the digital memory includes thermal cycle times and cycle numbers.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
- the microfluidic test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
- the microfluidic test module also has a photosensor wherein each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe- target hybrids being configured to emit photons in response to an input, and the photosensor is configured for sensing the photons emitted by the probe-target hybrids.
- the data stored in the digital memory includes hybridization data generated from the photosensor output.
- the microfluidic test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the controller is configured for transmission of the hybridization data to the dedicated reader.
- the sample is drawn from a patient and the controller is configured to download patient data via the dedicated reader and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the controller.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- GCA003-PCT The easily usable, mass-producible, inexpensive, compact, light, and portable Micro fluidic test module, with self-contained storage of required reagents, accepts a sample and processes and analyzes the sample material using the module's integral sensors, and provides the results electronically at its output port.
- the module's communications port is interfaced to an inexpensive and portable dedicated reader which provides the module with power, computing, communications, and user interface support.
- the dedicated reader obviates the need for heavy and expensive module support systems.
- microfluidic test module for analyzing a sample fluid and communicating test results to a desktop computer, the microfluidic test module comprising:
- a communication interface for communication with the desktop computer; and, a controller for operative control of the communication interface.
- the communication interface is a universal serial bus (USB) device driver for operative control of a USB plug in the test module for communication with an external device.
- USB universal serial bus
- the USB device driver is configured to draw power from the desktop computer to power the controller and the functional sections.
- the controller is further configured for operative control of the functional sections during processing and analysis of the sample.
- the controller is configured to download data via the desktop computer.
- the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing data relating to the reagent identities.
- the data stored in the digital memory is a unique identifier for microfluidic test module.
- the sample is a biological sample containing genetic material and one of the functional sections is a nucleic acid amplification section for amplifying nucleic acid sequences in the genetic material.
- the nucleic acid amplification section is a polymerase chain reaction (PCR) section and the data stored in the digital memory includes thermal cycle times and cycle numbers.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
- the micro fluidic test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
- the micro fluidic test module also has a photosensor wherein each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe- target hybrids being configured to emit photons in response to an input, and the photosensor is configured for sensing the photons emitted by the probe-target hybrids.
- the data stored in the digital memory includes hybridization data generated from the photosensor output.
- the micro fluidic test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the controller is configured for transmission of the hybridization data to the desktop computer.
- the sample is drawn from a patient and the controller is configured to download patient data via the desktop computer and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the controller.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the
- GCA003-PCT liquid and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the easily usable, mass-producible, inexpensive, compact, light, and portable Micro fluidic test module accepts a sample and processes and analyzes the sample material using the module's integral sensors, and provides the results electronically at its output port.
- the module's communications port is interfaced to a desktop PC which provides the module with power, computing, communications, and user interface support.
- Desktop PCs are widely available and are inexpensive, obviating the need for specialized, heavy, and expensive module support systems.
- micro fluidic test module for analyzing a sample fluid and communicating test results to an ebook reader, the micro fluidic test module comprising:
- a communication interface for communication with the ebook reader; and, a controller for operative control of the communication interface.
- the communication interface is a universal serial bus
- USB universal adapter
- the USB device driver is configured to draw power from the ebook reader to power the controller and the functional sections.
- the controller is further configured for operative control of the functional sections during processing and analysis of the sample.
- the controller is configured to download data via the ebook reader.
- the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing data relating to the reagent identities.
- the data stored in the digital memory is a unique identifier for microfluidic test module.
- the sample is a biological sample containing genetic material and one of the functional sections is a nucleic acid amplification section for amplifying nucleic acid sequences in the genetic material.
- the nucleic acid amplification section is a polymerase chain reaction (PCR) section and the data stored in the digital memory includes thermal cycle times and cycle numbers.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
- the micro fluidic test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
- the microfluidic test module also has a photosensor wherein each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe- target hybrids being configured to emit photons in response to an input, and the photosensor is configured for sensing the photons emitted by the probe-target hybrids.
- the data stored in the digital memory includes hybridization data generated from the photosensor output.
- the microfluidic test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the controller is configured for transmission of the hybridization data to the ebook reader.
- the sample is drawn from a patient and the controller is configured to download patient data via the ebook reader and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the controller.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the easily usable, mass-producible, inexpensive, compact, light, and portable Micro fluidic test module accepts a sample and processes and analyzes the sample material using the module's integral sensors, and provides the results electronically at its output port.
- the module's communications port is interfaced to a ebook reader which provides the module with power, computing, communications, and user interface support. Ebook readers are widely and inexpensively available, obviating the need for specialized, heavy, and expensive module support systems.
- microfluidic test module for analyzing a sample fluid and communicating test results to a tablet computer, the microfluidic test module comprising:
- a communication interface for communication with the tablet computer; and, a controller for operative control of the communication interface.
- the communication interface is a universal serial bus
- USB universal adapter
- the USB device driver is configured to draw power from the tablet computer to power the controller and the functional sections.
- the controller is further configured for operative control of the functional sections during processing and analysis of the sample.
- the controller is configured to download data via the tablet computer.
- the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing data relating to the reagent identities.
- the data stored in the digital memory is a unique identifier for microfluidic test module.
- the sample is a biological sample containing genetic material and one of the functional sections is a nucleic acid amplification section for amplifying nucleic acid sequences in the genetic material.
- the nucleic acid amplification section is a polymerase chain reaction (PCR) section and the data stored in the digital memory includes thermal cycle times and cycle numbers.
- PCR polymerase chain reaction
- the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
- the microfluidic test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
- the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
- the microfluidic test module also has a photosensor wherein each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe- target hybrids being configured to emit photons in response to an input, and the photosensor is configured for sensing the photons emitted by the probe-target hybrids.
- the data stored in the digital memory includes hybridization data generated from the photosensor output.
- the microfluidic test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
- the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
- the controller is configured for transmission of the hybridization data to the tablet computer.
- the sample is drawn from a patient and the controller is configured to download patient data via the tablet computer and store the patient data in the digital memory.
- the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the controller.
- the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
- the easily usable, mass-producible, inexpensive, compact, light, and portable Micro fluidic test module accepts a sample and processes and analyzes the sample material using the module's integral sensors, and provides the results electronically at its output port.
- the module's communications port is interfaced to a tablet computer which provides the module with power, computing, communications, and user interface support. Tablet computers are widely and inexpensively available, obviating the need for specialized, heavy, and expensive module support systems.
- GMV001.1 This aspect of the invention provides a reagent microvial for a reagent dispensing apparatus, the microvial comprising:
- a container for holding a volume of reagent for holding a volume of reagent
- a droplet generator for ejecting droplets of the reagent from the container; and, electrical contacts for connection to a control processor in the reagent dispensing apparatus to receive drive pulses for the droplet generator and provide the data to the control processor.
- the container holds between 282 microliters and 400 microliters of reagent.
- the droplet generator is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
- the droplet generator has a piezo-electric actuator.
- the data includes an identity transmitted to the control processor.
- the identity is a unique identity distinguishing the microvial from all other microvials.
- the data is encrypted.
- the reagent microvial with nonvolatile digital memory is used to receive a reagent, store it, and dispense it under digital control using a droplet generator.
- the droplet generator being part of the microvial provides for a self-contained and volumetrically and positionally precise reagent dispensing technique, simplifying the complexity, increasing the reliability, and reducing the cost of the automated manufacturing environment utilizing the microvial.
- the digital memory is used to store the information required during the functioning of the device in an automated manufacturing environment.
- the digital memory being part of the microvial, provides for an easily usable, reliable, safe, secure, and inexpensive approach to reagent dispensing in an automated manufacturing environment.
- This aspect of the invention provides a reagent microvial for a reagent dispensing apparatus, the microvial comprising:
- a container for holding a volume of reagent for holding a volume of reagent
- the microvial also has a droplet generator for ejecting droplets of the reagent from the container.
- the container holds between 282 microliters and 400 microliters of reagent.
- the droplet generator is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
- the droplet generator has a piezo-electric actuator.
- the identity is a unique identity distinguishing the microvial from all other microvials.
- the data is encrypted.
- the reagent microvial with authentication integrated circuit is used to receive a reagent, store it, and dispense it under digital control using a droplet generator.
- the droplet generator being part of the microvial provides for a self-contained and volumetrically and positionally precise reagent dispensing technique, simplifying the complexity, increasing the reliability, and reducing the cost of the automated manufacturing environment utilizing the microvial.
- the authentication integrated circuit is used to store the microvial authentication information used during the functioning of the device in an automated manufacturing environment.
- the authentication integrated circuit being part of the microvial, provides for an easily usable, reliable, safe, secure, and inexpensive approach to reagent dispensing in an automated manufacturing environment.
- GMV003.1 This aspect of the invention provides a reagent microvial for a reagent dispensing apparatus, the microvial comprising:
- a container for holding a volume of reagent for holding a volume of reagent
- the microvial also has a droplet generator for ejecting droplets of the reagent from the container.
- the container holds between 282 microliters and 400 microliters of reagent.
- the droplet generator is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
- the droplet generator has a piezo-electric actuator.
- the identity is a unique identity distinguishing the microvial from all other microvials.
- GCA003-PCT GMV003.7 Preferably, the data is encrypted.
- the reagent microvial with nonvolatile digital memory is used to receive a reagent, store it, and dispense it under digital control using a droplet generator.
- the droplet generator being part of the microvial provides for a self-contained and volumetrically and positionally precise reagent dispensing technique, simplifying the complexity, increasing the reliability, and reducing the cost of the automated manufacturing environment utilizing the microvial.
- the digital memory is used to store the information required during the functioning of the device in an automated manufacturing environment.
- the information stored in the memory includes the reagent specification data written into the memory by segments of the automated manufacturing environment. This information gets read from this memory an utilized as required by other segments of the automated manufacturing environment.
- the digital memory being part of the microvial, provides for an easily usable, reliable, safe, secure, and inexpensive approach to reagent dispensing in an automated manufacturing environment.
- GMV004.1 This aspect of the invention provides an oligonucleotide microvial for an oligonucleotide dispensing apparatus, the microvial comprising:
- a container for holding a volume of reagent for holding a volume of reagent
- the microvial also has a droplet generator for ejecting droplets of the oligonucleotides from the container.
- the container holds between 282 microliters and 400 microliters of reagent.
- the droplet generator is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
- the droplet generator has a piezo-electric actuator.
- the identity is a unique identity distinguishing the microvial from all other microvials.
- the data is encrypted.
- the oligonucleotide microvial with nonvolatile digital memory is used to receive a reagent, store it, and dispense it under digital control using a droplet generator.
- the droplet generator being part of the microvial provides for a self-contained and volumetrically and positionally precise oligonucleotide dispensing technique, simplifying the complexity, increasing the reliability, and reducing the cost of the automated manufacturing environment utilizing the microvial.
- the digital memory is used to store the information required during the functioning of the device in an automated manufacturing environment.
- the information stored in the memory includes the oligonucleotide specification data written into the memory by segments of the automated manufacturing environment. This information gets read from this memory an utilized as required by other segments of the automated manufacturing environment.
- the digital memory being part of the microvial, provides for an easily usable, reliable, safe, secure, and inexpensive approach to reagent dispensing in an automated manufacturing environment.
- This aspect of the invention provides a reagent dispensing apparatus for loading reagents into a microfluidic device having a digital memory for data related to the reagents loaded into the microfluidic device, the reagent dispensing apparatus comprising:
- reagent vials each of the vials having an integrated circuit with memory storing data regarding the reagent in the vial, and a droplet dispenser;
- a mounting surface for detachably mounting the microfluidic device for movement relative to the vials
- control processor for operative control of the vials and the mounting surface
- control processor is configured to activate the droplet dispenser of the vial selected, move the vial into registration with the microfluidic device and download the data from the integrated circuit to the digital memory of the microfluidic device.
- the reagent dispensing apparatus also has a camera for optical feedback of the registration between the vial selected by the control processor and the microfluidic device.
- the vial is a microvial for holding between 282 microliters and 400 microliters.
- the integrated circuit for each of the microvials has a unique identifier for identifying each of the microvials individually, the unique identifier being transmitted to the control processor and the digital memory of the microfluidic device.
- each of the microvials has electrical contacts for receiving activation pulses for the droplet dispenser and allowing the control processor to interrogate the integrated circuit.
- the reagent dispensing apparatus also has a rack wherein the microvials are detachably mounted to the rack for mechanical and electronic control of the microvials.
- the mounting surface is a stage configured for movement along two orthogonal axes, the rack extending parallel to one if the orthogonal axes.
- the microfluidic device is a lab-on-a-chip (LOC) device.
- LOC lab-on-a-chip
- the droplet dispenser has a piezo-electric actuator.
- the droplet dispenser is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
- the reagent dispensing apparatus also has facilities configured for applying a seal to the LOC device to close a plurality of reservoirs in which the reagents have been loaded.
- the LOC has a polymerase chain reaction (PCR) section and the list of reagents has one or more of:
- dNTPs deoxyribonucleoside triphosphates
- the reagent dispensing apparatus is used, as part of a cost-effective automated mass-manufacturing environment, to dispense reagents contained in reagent microvials into the reagent reservoirs of microfluidic devices.
- the data automation provided by the reagent dispensing apparatus includes automated computer-controlled dispensing of the reagents into the reagent reservoirs of the microfluidic devices, checking the reagent data stored in the memory of the microvials against the list of specifications for the reagents that have to be loaded in the microfluidic device, and storage of the reagent data into the memory of the microfluidic device.
- the reagent dispensing apparatus provides for an automated and volumetrically and positionally precise reagent dispensing technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
- the data automation provided by the reagent dispensing apparatus provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
- This aspect of the invention provides a reagent dispensing apparatus for loading reagents into a microfluidic device having a digital memory for data related to the reagents loaded into the microfluidic device, the reagent dispensing apparatus comprising:
- reagent vials each of the vials having an integrated circuit with memory storing data regarding the reagent in the vial, and a droplet dispenser;
- a mounting surface for detachably mounting the microfluidic device for movement relative to the vials
- control processor for operative control of the vials and the mounting surface
- control processor is configured to automatically interrogate each of the integrated circuits to collect and store the data regarding the reagents in each of the vials.
- control processor is configured to automatically activate the droplet dispenser of the vial selected, move the vial into registration with the
- GCA003-PCT microfluidic device download information from the integrated circuit to the digital memory of the microfluidic device.
- the reagent dispensing apparatus also has a camera for optical feedback of the registration between the vial selected by the control processor and the microfluidic device.
- the vial is a microvial for holding between 282 microliters and 400 microliters.
- the integrated circuit for each of the microvials has a unique identifier for identifying each of the microvials individually, the unique identifier being transmitted to the control processor and the digital memory of the microfluidic device.
- each of the microvials has electrical contacts for receiving activation pulses for the droplet dispenser and allowing the control processor to interrogate the integrated circuit.
- the reagent dispensing apparatus also has a rack wherein the microvials are detachably mounted to the rack for mechanical and electronic control of the microvials.
- the mounting surface is a stage configured for movement along two orthogonal axes, the rack extending parallel to one if the orthogonal axes.
- the microfluidic device is a lab-on-a-chip (LOC) device.
- LOC lab-on-a-chip
- the droplet dispenser has a piezo-electric actuator.
- the droplet dispenser is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
- the LOC has a polymerase chain reaction (PCR) section and the list of reagents has one or more of:
- dNTPs deoxyribonucleoside triphosphates
- the reagent dispensing apparatus is used, as part of a cost-effective automated mass-manufacturing environment, to dispense reagents contained in reagent microvials into the reagent reservoirs of microfluidic devices.
- the data automation provided by the reagent dispensing apparatus includes automated computer-controlled dispensing of the reagents into the reagent reservoirs of the microfluidic devices, checking the reagent data stored in the memory of the microvials against the list of specifications for the reagents that have to be loaded in the microfluidic device, and storage of the reagent data into the memory of the microfluidic device and in the reagent dispensing apparatus's computer memory.
- the reagent dispensing apparatus provides for an automated and volumetrically and positionally precise reagent dispensing technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
- the data automation provided by the reagent dispensing apparatus provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
- This aspect of the invention provides a reagent dispensing apparatus for loading reagents into a fixed array of microfluidic devices, each microfluidic device having a digital memory for data related to the reagents loaded into the microfluidic device, the reagent dispensing apparatus comprising:
- reagent vials each of the vials having an integrated circuit with memory storing data regarding the reagent in the vial, and a droplet dispenser;
- a mounting surface for detachably mounting the fixed array of microfluidic devices for movement relative to the vials
- control processor for operative control of the vials and the mounting surface
- control processor is configured to activate the droplet dispenser of the vial selected, move the vial into registration within one or more of the microfluidic devices within the fixed array and download the data from the integrated circuit to the digital memory of the one or more microfluidic devices.
- the fixed array of microfluidic devices is an array of lab- on-a-chip (LOC) devices mounted on a separable PCB (printed circuit board) wafer.
- LOC lab- on-a-chip
- the reagent dispensing apparatus also has a camera for optical feedback of the registration between the vial selected by the control processor and the LOC device.
- the vial is a microvial for holding between 282 microliters and 400 microliters.
- the integrated circuit for each of the microvials has a unique identifier for identifying each of the microvials individually, the unique identifier being transmitted to the control processor and the digital memory of the LOC device.
- each of the microvials has electrical contacts for receiving activation pulses for the droplet dispenser and allowing the control processor to interrogate the integrated circuit.
- the reagent dispensing apparatus also has a rack wherein the microvials are detachably mounted to the rack for mechanical and electronic control of the microvials.
- the mounting surface is a stage configured for movement along two orthogonal axes, the rack extending parallel to one if the orthogonal axes.
- the droplet dispenser has a piezo-electric actuator.
- the droplet dispenser is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
- the reagent dispensing apparatus also has facilities configured for applying a seal to the LOC device to close a plurality of reservoirs in which the reagents have been loaded.
- the LOC has a polymerase chain reaction (PCR) section and the list of reagents has one or more of:
- dNTPs deoxyribonucleoside triphosphates
- control processor is configured to automatically interrogate each of the integrated circuits to collect and store the data regarding the reagents in each of the vials.
- the reagent dispensing apparatus is used, as part of a cost-effective automated mass-manufacturing environment, to dispense reagents contained in reagent microvials into the reagent reservoirs of arrays of micro fluidic devices mounted on PCB wafers.
- the data automation provided by the reagent dispensing apparatus includes automated computer-controlled dispensing of the reagents into the reagent reservoirs of the micro fluidic devices, checking the reagent data stored in the memory of the microvials against the list of specifications for the reagents that have to be loaded in the microfluidic device, and storage of the reagent data into the memory of the microfluidic device.
- the reagent dispensing apparatus provides for an automated and volumetrically and positionally precise reagent dispensing technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
- the data automation provided by the reagent dispensing apparatus provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
- Dispensing of the reagents into the arrays of microfluidic devices mounted on PCB wafers speeds up and reduces the cost of the loading process, and by loading the reagents into the microfluidic devices after mounting the devices on the PCB wafer and soldering them, improves the chemical and physical integrity of the reagents.
- This aspect of the invention provides a reagent dispensing apparatus for loading reagents into a silicon wafer on which an array of lab-on-a-chip (LOC) devices are fabricated, each LOC device having a digital memory for data related to the reagents loaded into the LOC device, the reagent dispensing apparatus comprising:
- reagent vials each of the vials having an integrated circuit with memory storing data regarding the reagent in the vial, and a droplet dispenser;
- GCA003-PCT a mounting surface for detachably mounting the silicon wafer for movement relative to the vials;
- control processor for operative control of the vials and the mounting surface
- control processor is configured to activate the droplet dispenser of the vial selected, move the vial into registration within one or more of the LOC devices on the silicon wafer and download the data from the integrated circuit to the digital memory of the one or more LOC devices.
- the silicon wafer is partially sawn in preparation for tessellation into individually separate LOC devices.
- the reagent dispensing apparatus also has a camera for optical feedback of the registration between the vial selected by the control processor and the LOC device.
- the vial is a microvial for holding between 282 microliters and 400 microliters.
- the integrated circuit for each of the microvials has a unique identifier for identifying each of the microvials individually, the unique identifier being transmitted to the control processor and the digital memory of the LOC device.
- each of the microvials has electrical contacts for receiving activation pulses for the droplet dispenser and allowing the control processor to interrogate the integrated circuit.
- the reagent dispensing apparatus also has a rack wherein the microvials are detachably mounted to the rack for mechanical and electronic control of the microvials.
- the mounting surface is a stage configured for movement along two orthogonal axes, the rack extending parallel to one if the orthogonal axes.
- the droplet dispenser has a piezo-electric actuator.
- the droplet dispenser is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
- the reagent dispensing apparatus also has facilities configured for applying a seal to the LOC device to close a plurality of reservoirs in which the reagents have been loaded.
- the LOC has a polymerase chain reaction (PCR) section and the list of reagents has one or more of:
- dNTPs deoxyribonucleoside triphosphates
- control processor is configured to automatically interrogate each of the integrated circuits to collect and store the data regarding the reagents in each of the vials.
- the reagent dispensing apparatus is used, as part of a cost-effective automated mass-manufacturing environment, to dispense reagents contained in reagent microvials into the reagent reservoirs of micro fluidic devices on partial-depth sawn silicon wafers.
- the data automation provided by the reagent dispensing apparatus includes automated computer-controlled dispensing of the reagents into the reagent reservoirs of the micro fluidic devices, checking the reagent data stored in the memory of the microvials against the list of specifications for the reagents that have to be loaded in the microfluidic device, and storage of the reagent data into the memory of the microfluidic device.
- the reagent dispensing apparatus provides for an automated and volumetrically and positionally precise reagent dispensing technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
- the data automation provided by the reagent dispensing apparatus provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
- Dispensing of the reagents into microfluidic devices on partial-depth sawn silicon wafers speeds up the process of loading and reduces its cost.
- This aspect of the invention provides an oligonucleotide spotting device for contactless spotting of oligonucleotide probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the oligonucleotide spotting device comprising:
- each of the ejectors being configured for fluid communication with a corresponding one of the reservoirs respectively;
- the ejectors are configured to eject droplets containing the oligonucleotide probes from the corresponding reservoir onto the surface.
- each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
- the ejector has a chamber for containing the liquid from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
- the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
- the ejectors are configured to eject droplets having a volume less than 100 picoliters.
- the ejectors are configured to eject droplets having a volume less than 25 picoliters.
- the ejectors are configured to eject droplets having a volume less than 6 picoliters.
- the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
- the actuators in each of the ejectors are configured to actuate individually.
- each of the ejectors has a plurality of inlet channels extending from the reservoir to the chamber.
- the oligonucleotide spotting device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond- pads for connection to a microprocessor controller operatively controlling relative
- the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
- the supporting substrate has a reservoir side and an ejector side opposite the reservoir side wherein the array of reservoirs is formed in the reservoir side and the array of ejectors is formed on the ejector side.
- the CMOS circuitry is between the array of reservoirs and the array of ejectors.
- the array of ejectors is configured to eject droplets containing the oligonucleotide probes onto the surface with a density greater than 1 droplet per square millimeter.
- the array of ejectors is configured to eject droplets containing the oligonucleotide probes onto the surface with a density greater than 8 droplets per square millimeter.
- the array of ejectors is configured to eject droplets containing the oligonucleotide probes onto the surface with a density greater than 60 droplets per square millimeter.
- the array of ejectors is configured to eject droplets containing the oligonucleotide probes onto the surface with a density between 500 droplets per square millimeter and 1500 droplets per square millimeter.
- the array of ejectors is configured to eject droplets containing the oligonucleotide probes onto the surface at a rate greater than 100 droplets per second.
- the array of ejectors is configured to eject droplets containing the oligonucleotide probes onto the surface at a rate greater than 1,400 droplets per second.
- the mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted.
- the data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides onto the surface being spotted, receiving the specifications of the oligonucleotides stored in
- the GCA003-PCT its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment.
- the oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
- the device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
- the data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
- This aspect of the invention provides a spotting device for contactless spotting of a lab-on-a-chip (LOC) device with oligonucleotide probes, the LOC device having an array of hybridization chambers for receiving the oligonucleotide probes, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample and the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, the spotting device comprising:
- each of the ejectors being configured for fluid communication with a corresponding one of the reservoirs respectively such that the ejectors eject droplets containing the oligonucleotide probes from the corresponding reservoir into one of the hybridization chambers;
- the array of reservoirs is configured to contain the complete assay of
- oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
- the array of reservoirs has more than 1000 reservoirs.
- each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
- the ejector has a chamber for containing the liquid from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
- the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
- the ejectors are configured to eject droplets having a volume less than 100 picoliters.
- the ejectors are configured to eject droplets having a volume less than 25 picoliters.
- the ejectors are configured to eject droplets having a volume less than 6 picoliters.
- the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
- the actuators in each of the ejectors are configured to actuate individually.
- each of the ej ectors has a plurality of inlet channels extending from the reservoir to the chamber.
- the spotting device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond-pads for connection to a microprocessor controller operatively controlling relative movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
- the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
- the spotting device also has a supporting substrate having a reservoir side and an ejector side opposite the reservoir side wherein the array of reservoirs is formed in the reservoir side and the array of ejectors is formed on the ejector side.
- the array of ejectors is configured to spot the oligonucleotides onto the surface with a density greater than 1 probe spot per square millimeter.
- the array of ejectors is configured to spot the oligonucleotides onto the surface with a density greater than 8 probe spots per square millimeter.
- the array of ejectors is configured to spot the oligonucleotides onto the surface with a density greater than 60 probe spots per square millimeter.
- the array of ejectors is configured to spot the oligonucleotides onto the surface with a density between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
- the array of ejectors is configured to spot the oligonucleotides onto the surface at a rate greater than 100 probe spots per second.
- the array of ejectors is configured to spot the oligonucleotides onto the surface at a rate greater than 1,400 probe spots per second.
- the mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them into the hybridization chambers of LOC devices that are being spotted.
- the data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides into the hybridization chambers of the LOC devices, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the
- oligonucleotide specifications for storage into the memory of the LOC devices that are being spotted.
- the oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
- the device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
- the large numbers of oligonucleotide reservoirs and ejectors available on the oligonucleotide spotting device also provide for a one-step spotting of each LOC device.
- the data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
- This aspect of the invention provides a biochemical deposition device for contactless deposition of biochemicals on a surface, the biochemical deposition device comprising:
- each of the ejectors being configured for fluid communication with a corresponding one of the reservoirs respectively;
- the ejectors are configured to eject droplets containing the biochemical from the corresponding reservoir onto the surface.
- the biochemicals in the array of reservoirs are oligonucleotide probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample
- the surface is a lab-on-a- chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes.
- LOC lab-on-a- chip
- the array of hybridization chambers is configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
- the array of reservoirs has more than 1000 reservoirs.
- each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
- the ejector has a chamber for containing liquid with suspended oligonucleotide probes supplied from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
- the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
- the ejectors are configured to eject droplets having a volume less than 100 picoliters.
- the ejectors are configured to eject droplets having a volume less than 25 picoliters.
- the ejectors are configured to eject droplets having a volume less than 6 picoliters.
- the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
- the actuators in each of the ejectors are configured to actuate individually.
- each of the ejectors has a plurality of inlet channels extending from the reservoir to the chamber.
- the biochemical deposition device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond- pads for connection to a microprocessor controller operatively controlling relative movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
- the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
- the biochemical deposition device also has a supporting substrate having a reservoir side and an ejector side opposite the reservoir side wherein the array of reservoirs is formed in the reservoir side and the array of ejectors is formed on the ejector side.
- the array of ejectors is configured to eject droplets containing the biochemicals onto the surface with a density greater than 8 droplets per square millimeter.
- the array of ejectors is configured to eject droplets containing the biochemicals onto the surface with a density greater than 60 droplets per square millimeter.
- the array of ejectors is configured to eject droplets containing the biochemicals onto the surface with a density between 500 droplets per square millimeter and 1500 droplets per square millimeter.
- the array of ejectors is configured to eject droplets containing the biochemicals onto the surface at a rate greater than 100 droplets per second.
- the mass-producible and inexpensive biochemical deposition device is used as a part of a cost-effective automated mass-manufacturing environment.
- Biochemicals are loaded in the device's biochemical reservoirs, and the device deposits them onto a surface by ejecting the biochemicals from its biochemical reservoir onto the surfaces being deposited upon.
- the data automation provided by the biochemical deposition device includes automated computer-controlled dispensing of the biochemicals onto the surface being spotted, receiving the specifications of the biochemicals stored in its reservoirs, storing the biochemicals specifications in its digital memory, and transmitting of the biochemicals specifications to segments of the automated manufacturing environment.
- the biochemical deposition device provides for an automated, volumetrically and positionally precise, fast, and high-density biochemical deposition technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
- the device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
- the data automation provided by the biochemical deposition device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
- MST microsystems technology
- a monolithic substrate having a reservoir side and an ejector side opposite the reservoir side;
- the ejectors are configured to eject droplets containing the oligonucleotide probes from the corresponding reservoir onto the surface.
- each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
- the ejector has a chamber for containing liquid with suspended oligonucleotide probes supplied from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
- the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
- the ejectors are configured to eject droplets having a volume less than 100 picoliters.
- the ejectors are configured to eject droplets having a volume less than 25 picoliters.
- the ejectors are configured to eject droplets having a volume less than 6 picoliters.
- the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
- the actuators in each of the ejectors are configured to actuate individually.
- each of the ejectors has a plurality of inlet channels extending from the reservoir to the chamber.
- the MST device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond-pads for connection to a microprocessor controller operatively controlling relative movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
- the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
- the surface is a lab-on-a-chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes, the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of
- LOC lab-on-a-chip
- GCA003-PCT reservoirs being configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
- the array of reservoirs has more than 1000 reservoirs.
- the CMOS circuitry is between the array of reservoirs and the array of ej ectors .
- the array of ejectors is configured to spot the oligonucleotides onto the surface with a density greater than 8 probe spots per square millimeter.
- the array of ejectors is configured to spot the oligonucleotides onto the surface with a density greater than 60 probe spots per square millimeter.
- the array of ejectors is configured to spot the oligonucleotides onto the surface with a density between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
- the array of ejectors is configured to spot the oligonucleotides onto the surface at a rate greater than 100 probe spots per second.
- the array of ejectors is configured to spot the oligonucleotides onto the surface at a rate greater than 1,400 probe spots per second.
- the oligonucleotide spotting device is mass-produced inexpensively using microsystem technology (MST) and is used as a part of a cost-effective automated mass- manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted.
- MST microsystem technology
- the data automation provided by the oligonucleotide spotting device includes automated computer- controlled dispensing of the oligonucleotides onto the surface being spotted, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment.
- the oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
- the device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids
- GCA003-PCT from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
- the data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
- This aspect of the invention provides an oligonucleotide spotting device for contactless spotting of oligonucleotide probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the oligonucleotide spotting device comprising:
- the ejectors are configured to eject droplets containing the oligonucleotide probes from the corresponding reservoir onto the surface.
- the oligonucleotide spotting device also has a supporting substrate having a reservoir side and an ejector side opposite the reservoir side wherein the array of reservoirs is formed in the reservoir side and the array of ejectors is formed on the ejector side.
- each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
- the ejector has a chamber for containing the liquid from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
- the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
- the ejectors are configured to eject droplets having a volume less than 100 picoliters.
- the ejectors are configured to eject droplets having a volume less than 25 picoliters.
- the ejectors are configured to eject droplets having a volume less than 6 picoliters.
- the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
- the actuators in each of the ejectors are configured to actuate individually.
- each of the ejectors has a plurality of inlet channels extending from the reservoir to the chamber.
- the oligonucleotide spotting device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond- pads for connection to a microprocessor controller operatively controlling relative movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
- the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
- the CMOS circuitry is between the array of reservoirs and the array of ejectors.
- the surface is a lab-on-a-chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes, the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
- LOC lab-on-a-chip
- the array of reservoirs has more than 1000 reservoirs.
- the array of ejectors is configured to spot the probes onto the surface at a density more than 8 probe spots per square millimeter.
- the array of ejectors is configured to spot the probes onto the surface at a density more than 60 probe spots per square millimeter.
- the array of ejectors is configured to spot the probes onto the surface at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
- the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
- GCA003-PCT The mass-producible and inexpensive oligonucleotide spotting device with laminar structure is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted.
- the data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides onto the surfaces being spotted, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment.
- the oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
- the device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
- the data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
- This aspect of the invention provides an oligonucleotide spotting device for contactless spotting of oligonucleotide probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the oligonucleotide spotting device comprising:
- the reservoirs configured for containing the oligonucleotide probes suspended in a liquid
- the ejectors are configured to eject droplets containing the oligonucleotide probes from the corresponding reservoir onto the surface.
- each of the ejectors is configured for fluid
- each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
- the ejector has a chamber for containing the liquid from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
- the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
- the ejectors are configured to eject droplets having a volume less than 100 picoliters.
- the ejectors are configured to eject droplets having a volume less than 25 picoliters.
- the ejectors are configured to eject droplets having a volume less than 6 picoliters.
- the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
- the actuators in each of the ejectors are configured to actuate individually.
- each of the ejectors is in fluid communication with one of the reservoirs via more than one of the inlet channels.
- the oligonucleotide spotting device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond- pads for connection to a microprocessor controller operatively controlling relative movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
- the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
- the CMOS circuitry is between the array of reservoirs and the array of ejectors.
- the array of ejectors is configured to spot the probes onto the surface at a density more than 8 probe spots per square millimeter.
- the array of ejectors is configured to spot the probes onto the surface at a density more than 60 probe spots per square millimeter.
- the array of ejectors is configured to spot the probes onto the surface at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
- the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
- the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 1,400 probe spots per second.
- the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 20,000 probe spots per second.
- the mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. It is fabricated with fluidics on both side of a silicon substrate, increasing the device integration level, reducing the device dimensions, and minimizing the device cost. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted.
- the data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides onto the surface being spotted, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment.
- the oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
- the device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids
- GCA003-PCT from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
- the data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
- This aspect of the invention provides an oligonucleotide spotting device for contactless spotting of probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the oligonucleotide spotting device comprising:
- CMOS circuitry for providing each of the drop ejection actuators with a drive pulse for droplet ejection
- bond-pads for electrically connecting the CMOS circuitry and an external microprocessor controller for operative control of the array of ejectors.
- the CMOS circuitry has a digital memory storing identity data for identifying the device to the external microprocessor controller.
- the oligonucleotide spotting device also has an array of reservoirs for containing the oligonucleotide probes suspended in a liquid, wherein the supporting substrate has a reservoir side and an ejector side opposite the reservoir side, the array of reservoirs being formed in the reservoir side and, the array of ejectors formed on the ejector side, each of the ejectors being configured for fluid communication with a corresponding one of the probe reservoirs respectively.
- the digital memory stores specification data for the oligonucleotide probes.
- each of the ejectors has a plurality of the actuators and a corresponding plurality of nozzles associated with each of the droplet ejection actuators respectively, such that actuation of one of the actuators ejects a droplet through the nozzle associated with said actuator.
- the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
- the ejectors are configured to eject droplets having a volume less than 100 picoliters.
- the ejectors are configured to eject droplets having a volume less than 25 picoliters.
- the ejectors are configured to eject droplets having a volume less than 6 picoliters.
- the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
- the actuators in one of the ejectors are configured to actuate individually.
- each of the ejectors have a chamber for containing the liquid for ejection from the nozzle, and a plurality of inlet channels extending from the chamber to the reservoir corresponding to the ejector.
- the actuators in each of the ejectors are configured to actuate individually.
- the CMOS circuitry is between the array of reservoirs and the array of ejectors.
- the surface is a lab-on-a-chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes, the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
- LOC lab-on-a-chip
- the array of reservoirs has more than 1000 reservoirs.
- the array of ejectors is configured to spot the probes onto the surface at a density more than 8 probe spots per square millimeter.
- the array of ejectors is configured to spot the probes onto the surface at a density more than 60 probe spots per square millimeter.
- the array of ejectors is configured to spot the probes onto the surface at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
- the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
- the mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted.
- the data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides onto the surface being spotted, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment. The spotting device performs these functions under external computer control.
- the oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
- the device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
- the data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
- GPD009.1 This aspect of the invention provides an oligonucleotide spotting device for contactless spotting of probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the oligonucleotide spotting device comprising:
- CMOS circuitry for providing each of the drop ejection actuators with a drive pulse for droplet ejection
- the CMOS circuitry has a digital memory for storing data related to the device.
- the CMOS circuitry has bond-pads for electrically connecting and an external microprocessor controller for operative control of the array of ejectors.
- the data includes identity data for identifying the device to the external microprocessor controller.
- the oligonucleotide spotting device also has an array of reservoirs for containing the oligonucleotide probes suspended in a liquid, wherein the supporting substrate has a reservoir side and an ejector side opposite the reservoir side, the array of reservoirs being formed in the reservoir side and, the array of ejectors formed on the ejector side, each of the ejectors being configured for fluid communication with a corresponding one of the probe reservoirs respectively.
- the digital memory stores specification data for the oligonucleotide probes.
- each of the ejectors has a plurality of the actuators and a corresponding plurality of nozzles associated with each of the droplet ejection actuators respectively, such that actuation of one of the actuators ejects a droplet through the nozzle associated with said actuator.
- the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
- the ejectors are configured to eject droplets having a volume less than 100 picoliters.
- the ejectors are configured to eject droplets having a volume less than 25 picoliters.
- the ejectors are configured to eject droplets having a volume less than 6 picoliters.
- the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
- the actuators in one of the ejectors are configured to actuate individually.
- each of the ejectors have a chamber for containing the liquid for ejection from the nozzle, and a plurality of inlet channels extending from the chamber to the reservoir corresponding to the ejector.
- the actuators in each of the ejectors are configured to actuate individually.
- the CMOS circuitry is between the array of reservoirs and the array of ejectors.
- the surface is a lab-on-a-chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes, the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
- LOC lab-on-a-chip
- the array of reservoirs has more than 1000 reservoirs.
- the array of ejectors is configured to spot the probes onto the surface at a density more than 60 probe spots per square millimeter.
- the array of ejectors is configured to spot the probes onto the surface at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
- the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
- the mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted.
- the data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides onto the surface being spotted, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment.
- the oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
- the device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids
- GCA003-PCT from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
- the data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
- This aspect of the invention provides a spotting device for contactless spotting of oligonucleotide probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the spotting device comprising:
- CMOS circuitry for providing each of the drop ejection actuators with a drive pulse for droplet ejection
- the CMOS circuitry has a digital memory.
- the CMOS circuitry has bond-pads for electrically connecting and an external microprocessor controller for operative control of the array of ejectors.
- the digital memory stores identity data for identifying the device to the external microprocessor controller.
- the oligonucleotide spotting device also has a supporting substrate with a reservoir side and an ejector side opposite the reservoir side, the array of reservoirs being formed in the reservoir side and, the array of ejectors formed on the ejector side, each of the ejectors being configured for fluid communication with a corresponding one of the probe reservoirs respectively.
- the digital memory stores specification data for the oligonucleotide probes.
- each of the ejectors has a plurality of the actuators and a corresponding plurality of nozzles associated with each of the droplet ejection actuators respectively, such that actuation of one of the actuators ejects a droplet through the nozzle associated with said actuator.
- the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
- the ejectors are configured to eject droplets having a volume less than 2.0 picoliters.
- the actuators in one of the ejectors are configured to actuate individually.
- each of the ejectors have a chamber for containing the liquid for ejection from the nozzle, and a plurality of inlet channels extending from the chamber to the reservoir corresponding to the ejector.
- the actuators in each of the ejectors are configured to actuate individually.
- the CMOS circuitry is between the array of reservoirs and the array of ejectors.
- the surface is a lab-on-a-chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes, the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
- LOC lab-on-a-chip
- the array of reservoirs has more than 1000 reservoirs.
- the array of ejectors is configured to spot the probes onto the surface at a density more than 8 probe spots per square millimeter.
- the array of ejectors is configured to spot the probes onto the surface at a density more than 60 probe spots per square millimeter.
- the array of ejectors is configured to spot the probes onto the surface at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
- the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
- the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 1,400 probe spots per second.
- the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 20,000 probe spots per second.
- the mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted.
- the data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides onto the surface being spotted, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment.
- the oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
- the device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
- the data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
- This aspect of the invention provides a spotting device for contactless spotting of lab-on-a-chip (LOC) devices with oligonucleotide probes, the LOC devices being held in a fixed array on a printed circuit board (PCB) and each having an array of hybridization chambers for receiving the oligonucleotide probes, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the spotting device comprising:
- GCA003-PCT an array of reservoirs on one side of the supporting substrate, the reservoirs containing sufficient amount of the oligonucleotide probes suspended in a liquid to spot all the LOC devices on the PCB;
- each of the ejectors being configured for fluid communication with a corresponding one of the reservoirs respectively such that the ejectors eject droplets containing the oligonucleotide probes from the corresponding reservoir into one of the hybridization chambers.
- the array of reservoirs has more than 1000 reservoirs.
- each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
- the ejector has a chamber for containing the liquid from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
- the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
- the ejectors are configured to eject droplets having a volume less than 100 picoliters.
- the ejectors are configured to eject droplets having a volume less than 25 picoliters.
- the ejectors are configured to eject droplets having a volume less than 6 picoliters.
- the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
- the actuators in each of the ejectors are configured to actuate individually.
- each of the ej ectors has a plurality of inlet channels extending from the reservoir to the chamber.
- the spotting device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond-pads for connection to a microprocessor controller operatively controlling relative movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
- the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
- the array of reservoirs is integrally formed into the one side of the monolithic supporting substrate.
- the array of ejectors is configured to spot the probes onto the surface at a density more than 8 probe spots per square millimeter.
- the array of ejectors is configured to spot the probes onto the surface at a density more than 60 probe spots per square millimeter.
- the array of ejectors is configured to spot the probes onto the surface at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
- the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
- the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 1,400 probe spots per second.
- the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 20,000 probe spots per second.
- the mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them into the hybridization chambers of the arrays of LOC devices mounted on PCB wafers.
- the data automation provided by the oligonucleotide spotting device includes automated computer- controlled spotting with oligonucleotide of the arrays of LOC devices mounted on PCB wafers, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to the LOC devices or other segments of the automated manufacturing environment.
- the oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
- the device also functions
- GCA003-PCT as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
- the data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
- This aspect of the invention provides an oligonucleotide spotting device for contactless spotting of a silicon wafer on which an array of lab-on-a-chip (LOC) devices are fabricated, the LOC devices being configured to use the oligonucleotide probes to detect target nucleic acid sequences in a biological sample and each having an array of hybridization chambers for receiving the oligonucleotide probes, the oligonucleotide spotting device comprising:
Abstract
An apparatus for loading oligonucleotide spotting devices and spotting oligonucleotide probes, the apparatus having a plurality of oligonucleotide vials, each with a droplet dispenser, a mounting surface for detachably mounting an oligonucleotide spotting device, a chuck for detachably mounting the oligonucleotide spotting device adjacent the mounting surface, and, a control processor for operative control of the oligonucleotide vials, the oligonucleotide spotting device when mounted in the chuck and movement of the mounting surface relative to the oligonucleotide vials, and the oligonucleotide spotting device, wherein, the control processor is configured to activate the droplet dispensers, and move the oligonucleotide spotting device into registration with the oligonucleotide vials.
Description
APPARATUS FOR LOADING OLIGONUCLEOTIDE SPOTTING DEVICES AND SPOTTING OLIGONUCLEOTIDE PROBES
FIELD OF THE INVENTION
The present invention relates to diagnostic devices that use microsystems technologies (MST). In particular, the invention relates to microfluidic and biochemical processing and analysis for molecular diagnostics.
BACKGROUND OF THE INVENTION
Molecular diagnostics has emerged as a field that offers the promise of early disease detection, potentially before symptoms have manifested. Molecular diagnostic testing is used to detect:
• Inherited disorders
• Acquired disorders
• Infectious diseases
• Genetic predisposition to health-related conditions.
With high accuracy and fast turnaround times, molecular diagnostic tests have the potential to reduce the occurrence of ineffective health care services, enhance patient outcomes, improve disease management and individualize patient care. Many of the techniques in molecular diagnostics are based on the detection and identification of specific nucleic acids, both deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), extracted and amplified from a biological specimen (such as blood or saliva). The complementary nature of the nucleic acid bases allows short sequences of synthesized DNA (oligonucleotides) to bond (hybridize) to specific nucleic acid sequences for use in nucleic acid tests. If hybridization occurs, then the complementary sequence is present in the sample. This makes it possible, for example, to predict the disease a person will contract in the future, determine the identity and virulence of an infectious pathogen, or determine the response a person will have to a drug.
NUCLEIC ACID BASED MOLECULAR DIAGNOSTIC TEST
A nucleic acid based test has four distinct steps:
1. Sample preparation
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2. Nucleic acid extraction
3. Nucleic acid amplification (optional)
4. Detection Many sample types are used for genetic analysis, such as blood, urine, sputum and tissue samples. The diagnostic test determines the type of sample required as not all samples are representative of the disease process. These samples have a variety of constituents, but usually only one of these is of interest. For example, in blood, high concentrations of erythrocytes can inhibit the detection of a pathogenic organism.
Therefore a purification and/or concentration step at the beginning of the nucleic acid test is often required.
Blood is one of the more commonly sought sample types. It has three major constituents: leukocytes (white blood cells), erythrocytes (red blood cells) and
thrombocytes (platelets). The thrombocytes facilitate clotting and remain active in vitro. To inhibit coagulation, the specimen is mixed with an agent such as
ethylenediaminetetraacetic acid (EDTA) prior to purification and concentration.
Erythrocytes are usually removed from the sample in order to concentrate the target cells. In humans, erythrocytes account for approximately 99% of the cellular material but do not carry DNA as they have no nucleus. Furthermore, erythrocytes contain components such as haemoglobin that can interfere with the downstream nucleic acid amplification process (described below). Removal of erythrocytes can be achieved by differentially lysing the erythrocytes in a lysis solution, leaving remaining cellular material intact which can then be separated from the sample using centrifugation. This provides a concentration of the target cells from which the nucleic acids are extracted.
The exact protocol used to extract nucleic acids depends on the sample and the diagnostic assay to be performed. For example, the protocol for extracting viral RNA will vary considerably from the protocol to extract genomic DNA. However, extracting nucleic acids from target cells usually involves a cell lysis step followed by nucleic acid purification. The cell lysis step disrupts the cell and nuclear membranes, releasing the genetic material. This is often accomplished using a lysis detergent, such as sodium dodecyl sulfate, which also denatures the large amount of proteins present in the cells.
The nucleic acids are then purified with an alcohol precipitation step, usually ice- cold ethanol or isopropanol, or via a solid phase purification step, typically on a silica matrix in a column, resin or on paramagnetic beads in the presence of high concentrations
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of a chaotropic salt, prior to washing and then elution in a low ionic strength buffer. An optional step prior to nucleic acid precipitation is the addition of a protease which digests the proteins in order to further purify the sample.
Other lysis methods include mechanical lysis via ultrasonic vibration and thermal lysis where the sample is heated to 94°C to disrupt cell membranes.
The target DNA or RNA may be present in the extracted material in very small amounts, particularly if the target is of pathogenic origin. Nucleic acid amplification provides the ability to selectively amplify (that is, replicate) specific targets present in low concentrations to detectable levels.
The most commonly used nucleic acid amplification technique is the polymerase chain reaction (PC ). PCR is well known in this field and comprehensive description of this type of reaction is provided in E. van Pelt-Verkuil et al, Principles and Technical Aspects of PCR Amplification, Springer, 2008.
PCR is a powerful technique that amplifies a target DNA sequence against a background of complex DNA. If RNA is to be amplified (by PCR), it must be first transcribed into cDNA (complementary DNA) using an enzyme called reverse transcriptase. Afterwards, the resulting cDNA is amplified by PCR.
PCR is an exponential process that proceeds as long as the conditions for sustaining the reaction are acceptable. The components of the reaction are:
1. pair of primers - short single strands of DNA with around 10-30 nucleotides complementary to the regions flanking the target sequence
2. DNA polymerase - a thermostable enzyme that synthesizes DNA
3. deoxyribonucleoside triphosphates (dNTPs) - provide the nucleotides that are incorporated into the newly synthesized DNA strand
4. buffer - to provide the optimal chemical environment for DNA synthesis
PCR typically involves placing these reactants in a small tube (-10-50 microlitres) containing the extracted nucleic acids. The tube is placed in a thermal cycler; an instrument that subjects the reaction to a series of different temperatures for varying amounts of time. The standard protocol for each thermal cycle involves a denaturation phase, an annealing phase, and an extension phase. The extension phase is sometimes referred to as the primer extension phase. In addition to such three-step protocols, two-step thermal protocols can be employed, in which the annealing and extension phases are combined. The denaturation phase typically involves raising the temperature of the
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reaction to 90 - 95°C to denature the DNA strands; in the annealing phase, the temperature is lowered to ~50-60°C for the primers to anneal; and then in the extension phase the temperature is raised to the optimal DNA polymerase activity temperature of 60- 72°C for primer extension. This process is repeated cyclically around 20-40 times, the end result being the creation of millions of copies of the target sequence between the primers.
There are a number of variants to the standard PC protocol such as multiplex PCR, linker-primed PCR, direct PCR, tandem PCR, real-time PCR and reverse- transcriptase PCR, amongst others, which have been developed for molecular diagnostics.
Multiplex PCR uses multiple primer sets within a single PCR mixture to produce amplicons of varying sizes that are specific to different DNA sequences. By targeting multiple genes at once, additional information may be gained from a single test-run that otherwise would require several experiments. Optimization of multiplex PCR is more difficult though and requires selecting primers with similar annealing temperatures, and amplicons with similar lengths and base composition to ensure the amplification efficiency of each amplicon is equivalent.
Linker-primed PCR, also known as ligation adaptor PCR, is a method used to enable nucleic acid amplification of essentially all DNA sequences in a complex DNA mixture without the need for target-specific primers. The method firstly involves digesting the target DNA population with a suitable restriction endonuclease (enzyme). Double- stranded oligonucleotide linkers (also called adaptors) with a suitable overhanging end are then ligated to the ends of target DNA fragments using a ligase enzyme. Nucleic acid amplification is subsequently performed using oligonucleotide primers which are specific for the linker sequences. In this way, all fragments of the DNA source which are flanked by linker oligonucleotides can be amplified.
Direct PCR describes a system whereby PCR is performed directly on a sample without any, or with minimal, nucleic acid extraction. It has long been accepted that PCR reactions are inhibited by the presence of many components of unpurified biological samples, such as the haem component in blood. Traditionally, PCR has required extensive purification of the target nucleic acid prior to preparation of the reaction mixture. With appropriate changes to the chemistry and sample concentration, however, it is possible to perform PCR with minimal DNA purification, or direct PCR. Adjustments to the PCR chemistry for direct PCR include increased buffer strength, the use of polymerases which have high activity and processivity, and additives which chelate with potential polymerase inhibitors.
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Tandem PCR utilises two distinct rounds of nucleic acid amplification to increase the probability that the correct amplicon is amplified. One form of tandem PCR is nested PCR in which two pairs of PCR primers are used to amplify a single locus in separate rounds of nucleic acid amplification. The first pair of primers hybridize to the nucleic acid sequence at regions external to the target nucleic acid sequence. The second pair of primers (nested primers) used in the second round of amplification bind within the first PCR product and produce a second PCR product containing the target nucleic acid, that will be shorter than the first one. The logic behind this strategy is that if the wrong locus were amplified by mistake during the first round of nucleic acid amplification, the probability is very low that it would also be amplified a second time by a second pair of primers and thus ensures specificity.
Real-time PCR, or quantitative PCR, is used to measure the quantity of a PCR product in real time. By using a fluorophore-containing probe or fluorescent dyes along with a set of standards in the reaction, it is possible to quantitate the starting amount of nucleic acid in the sample. This is particularly useful in molecular diagnostics where treatment options may differ depending on the pathogen load in the sample.
Reverse-transcriptase PCR (RT-PCR) is used to amplify DNA from RNA. Reverse transcriptase is an enzyme that reverse transcribes RNA into complementary DNA (cDNA), which is then amplified by PCR. RT-PCR is widely used in expression profiling, to determine the expression of a gene or to identify the sequence of an RNA transcript, including transcription start and termination sites. It is also used to amplify RNA viruses such as human immunodeficiency virus or hepatitis C virus.
Isothermal amplification is another form of nucleic acid amplification which does not rely on the thermal denaturation of the target DNA during the amplification reaction and hence does not require sophisticated machinery. Isothermal nucleic acid amplification methods can therefore be carried out in primitive sites or operated easily outside of a laboratory environment. A number of isothermal nucleic acid amplification methods have been described, including Strand Displacement Amplification, Transcription Mediated Amplification, Nucleic Acid Sequence Based Amplification, Recombinase Polymerase Amplification, Rolling Circle Amplification, Ramification Amplification, Helicase-
Dependent Isothermal DNA Amplification and Loop-Mediated Isothermal Amplification.
Isothermal nucleic acid amplification methods do not rely on the continuing heat denaturation of the template DNA to produce single stranded molecules to serve as templates for further amplification, but instead rely on alternative methods such as
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enzymatic nicking of DNA molecules by specific restriction endonucleases, or the use of an enzyme to separate the DNA strands, at a constant temperature.
Strand Displacement Amplification (SDA) relies on the ability of certain restriction enzymes to nick the unmodified strand of hemi-modified DNA and the ability of a 5 '-3' exonuclease-deficient polymerase to extend and displace the downstream strand.
Exponential nucleic acid amplification is then achieved by coupling sense and antisense reactions in which strand displacement from the sense reaction serves as a template for the antisense reaction. The use of nickase enzymes which do not cut DNA in the traditional manner but produce a nick on one of the DNA strands, such as N. Alwl, N. BstNBl and Mly 1, are useful in this reaction. SDA has been improved by the use of a combination of a heat-stable restriction enzyme (Aval) and heat-stable Exo- polymerase (Bst polymerase). This combination has been shown to increase amplification efficiency of the reaction from 108 fold amplification to 1010 fold amplification so that it is possible using this technique to amplify unique single copy molecules.
Transcription Mediated Amplification (TMA) and Nucleic Acid Sequence Based
Amplification (NASBA) use an RNA polymerase to copy RNA sequences but not corresponding genomic DNA. The technology uses two primers and two or three enzymes, RNA polymerase, reverse transcriptase and optionally RNase H (if the reverse transcriptase does not have RNase activity). One primer contains a promoter sequence for RNA polymerase. In the first step of nucleic acid amplification, this primer hybridizes to the target ribosomal RNA (rRNA) at a defined site. Reverse transcriptase creates a DNA copy of the target rRNA by extension from the 3' end of the promoter primer. The RNA in the resulting RNA:DNA duplex is degraded by the RNase activity of the reverse transcriptase if present or the additional RNase H. Next, a second primer binds to the DNA copy. A new strand of DNA is synthesized from the end of this primer by reverse transcriptase, creating a double-stranded DNA molecule. RNA polymerase recognizes the promoter sequence in the DNA template and initiates transcription. Each of the newly synthesized RNA amplicons re-enters the process and serves as a template for a new round of replication.
In Recombinase Polymerase Amplification (RPA), the isothermal amplification of specific DNA fragments is achieved by the binding of opposing oligonucleotide primers to template DNA and their extension by a DNA polymerase. Heat is not required to denature the double-stranded DNA (dsDNA) template. Instead, RPA employs recombinase-primer complexes to scan dsDNA and facilitate strand exchange at cognate sites. The resulting
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structures are stabilised by single-stranded DNA binding proteins interacting with the displaced template strand, thus preventing the ejection of the primer by branch migration. Recombinase disassembly leaves the 3' end of the oligonucleotide accessible to a strand displacing DNA polymerase, such as the large fragment of Bacillus subtilis Pol I (Bsu), and primer extension ensues. Exponential nucleic acid amplification is accomplished by the cyclic repetition of this process.
Helicase-dependent amplification (HDA) mimics the in vivo system in that it uses a DNA helicase enzyme to generate single-stranded templates for primer hybridization and subsequent primer extension by a DNA polymerase. In the first step of the HDA reaction, the helicase enzyme traverses along the target DNA, disrupting the hydrogen bonds linking the two strands which are then bound by single-stranded binding proteins. Exposure of the single-stranded target region by the helicase allows primers to anneal. The DNA polymerase then extends the 3 ' ends of each primer using free deoxyribonucleoside triphosphates (dNTPs) to produce two DNA replicates. The two replicated dsDNA strands independently enter the next cycle of HDA, resulting in exponential nucleic acid amplification of the target sequence.
Other DNA-based isothermal techniques include Rolling Circle Amplification (RCA) in which a DNA polymerase extends a primer continuously around a circular DNA template, generating a long DNA product that consists of many repeated copies of the circle. By the end of the reaction, the polymerase generates many thousands of copies of the circular template, with the chain of copies tethered to the original target DNA. This allows for spatial resolution of target and rapid nucleic acid amplification of the signal. Up to 1012 copies of template can be generated in 1 hour. Ramification amplification is a variation of RCA and utilizes a closed circular probe (C-probe) or padlock probe and a DNA polymerase with a high processivity to exponentially amplify the C-probe under isothermal conditions.
Loop-mediated isothermal amplification (LAMP), offers high selectivity and employs a DNA polymerase and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA. An inner primer containing sequences of the sense and antisense strands of the target DNA initiates LAMP. The following strand displacement DNA synthesis primed by an outer primer releases a single-stranded DNA. This serves as template for DNA synthesis primed by the second inner and outer primers that hybridize to the other end of the target, which produces a stem-loop DNA structure. In subsequent LAMP cycling one inner primer hybridizes to the loop on the product and
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initiates displacement DNA synthesis, yielding the original stem-loop DNA and a new stem-loop DNA with a stem twice as long. The cycling reaction continues with accumulation of 109 copies of target in less than an hour. The final products are stem-loop DNAs with several inverted repeats of the target and cauliflower-like structures with multiple loops formed by annealing between alternately inverted repeats of the target in the same strand.
After completion of the nucleic acid amplification, the amplified product must be analysed to determine whether the anticipated amplicon (the amplified quantity of target nucleic acids) was generated. The methods of analyzing the product range from simply determining the size of the amplicon through gel electrophoresis, to identifying the nucleotide composition of the amplicon using DNA hybridization.
Gel electrophoresis is one of the simplest ways to check whether the nucleic acid amplification process generated the anticipated amplicon. Gel electrophoresis uses an electric field applied to a gel matrix to separate DNA fragments. The negatively charged DNA fragments will move through the matrix at different rates, determined largely by their size. After the electrophoresis is complete, the fragments in the gel can be stained to make them visible. Ethidium bromide is a commonly used stain which fluoresces under UV light.
The size of the fragments is determined by comparison with a DNA size marker (a DNA ladder), which contains DNA fragments of known sizes, run on the gel alongside the amplicon. Because the oligonucleotide primers bind to specific sites flanking the target DNA, the size of the amplified product can be anticipated and detected as a band of known size on the gel. To be certain of the identity of the amplicon, or if several amplicons have been generated, DNA probe hybridization to the amplicon is commonly employed.
DNA hybridization refers to the formation of double-stranded DNA by
complementary base pairing. DNA hybridization for positive identification of a specific amplification product requires the use of a DNA probe around 20 nucleotides in length. If the probe has a sequence that is complementary to the amplicon (target) DNA sequence, hybridization will occur under favourable conditions of temperature, pH and ionic concentration. If hybridization occurs, then the gene or DNA sequence of interest was present in the original sample.
Optical detection is the most common method to detect hybridization. Either the amplicons or the probes are labelled to emit light through fluorescence or
electrochemiluminescence. These processes differ in the means of producing excited states
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of the light-producing moieties, but both enable covalent labelling of nucleotide strands. In electrochemiluminescence (ECL), light is produced by luminophore molecules or complexes upon stimulation with an electric current. In fluorescence, it is illumination with excitation light which leads to emission.
Fluorescence is detected using an illumination source which provides excitation light at a wavelength absorbed by the fluorescent molecule, and a detection unit. The detection unit comprises a photosensor (such as a photomultiplier tube or charge-coupled device (CCD) array) to detect the emitted signal, and a mechanism (such as a wavelength- selective filter) to prevent the excitation light from being included in the photosensor output. The fluorescent molecules emit Stokes -shifted light in response to the excitation light, and this emitted light is collected by the detection unit. Stokes shift is the frequency difference or wavelength difference between emitted light and absorbed excitation light.
ECL emission is detected using a photosensor which is sensitive to the emission wavelength of the ECL species being employed. For example, transition metal-ligand complexes emit light at visible wavelengths, so conventional photodiodes and CCDs are employed as photosensors. An advantage of ECL is that, if ambient light is excluded, the ECL emission can be the only light present in the detection system, which improves sensitivity.
Microarrays allow for hundreds of thousands of DNA hybridization experiments to be performed simultaneously. Microarrays are powerful tools for molecular diagnostics with the potential to screen for thousands of genetic diseases or detect the presence of numerous infectious pathogens in a single test. A microarray consists of many different DNA probes immobilized as spots on a substrate. The target DNA (amplicon) is first labelled with a fluorescent or luminescent molecule (either during or after nucleic acid amplification) and then applied to the array of probes. The microarray is incubated in a temperature controlled, humid environment for a number of hours or days while hybridization between the probe and amplicon takes place. Following incubation, the microarray must be washed in a series of buffers to remove unbound strands. Once washed, the microarray surface is dried using a stream of air (often nitrogen). The stringency of the hybridization and washes is critical. Insufficient stringency can result in a high degree of nonspecific binding. Excessive stringency can lead to a failure of appropriate binding, which results in diminished sensitivity. Hybridization is recognized by detecting light emission from the labelled amplicons which have formed a hybrid with complementary probes.
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Fluorescence from microarrays is detected using a microarray scanner which is generally a computer controlled inverted scanning fluorescence confocal microscope which typically uses a laser for excitation of the fluorescent dye and a photosensor (such as a photomultiplier tube or CCD) to detect the emitted signal. The fluorescent molecules emit Stokes-shifted light (described above) which is collected by the detection unit.
The emitted fluorescence must be collected, separated from the unabsorbed excitation wavelength, and transported to the detector. In microarray scanners, a confocal arrangement is commonly used to eliminate out-of- focus information by means of a confocal pinhole situated at an image plane. This allows only the in- focus portion of the light to be detected. Light from above and below the plane of focus of the object is prevented from entering the detector, thereby increasing the signal to noise ratio. The detected fluorescent photons are converted into electrical energy by the detector which is subsequently converted to a digital signal. This digital signal translates to a number representing the intensity of fluorescence from a given pixel. Each feature of the array is made up of one or more such pixels. The final result of a scan is an image of the array surface. The exact sequence and position of every probe on the microarray is known, and so the hybridized target sequences can be identified and analysed simultaneously.
More information regarding fluorescent probes can be found at:
http://www. premierbiosoft. com/tech_notes/FRET_probe. html and
http://www. invitrogen. com/site/us/en/nome/References/Molecular-Probes-The- Handbook/Technical-Notes-and-Product-Highlights/Fluorescence-Resonance- Energy-Transfer-FRET. html
POINT-OF-CARE MOLECULAR DIAGNOSTICS
Despite the advantages that molecular diagnostic tests offer, the growth of this type of testing in the clinical laboratory has been slower than expected and remains a minor part of the practice of laboratory medicine. This is primarily due to the complexity and costs associated with nucleic acid testing compared with tests based on methods not involving nucleic acids. The widespread adaptation of molecular diagnostics testing to the clinical setting is intimately tied to the development of instrumentation that significantly reduces the cost, provides a rapid and automated assay from start (specimen processing) to finish (generating a result) and operates without major intervention by personnel.
A point-of-care technology serving the physician's office, the hospital bedside or even consumer-based, at home, would offer many advantages including:
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• rapid availability of results enabling immediate facilitation of treatment and improved quality of care.
• ability to obtain laboratory values from testing very small samples.
• reduced clinical workload.
· reduced laboratory workload and improved office efficiency by reducing
administrative work.
• improved cost per patient through reduced length of stay of hospitalization,
conclusion of outpatient consultation at the first visit, and reduced handling, storing and shipping of specimens.
· facilitation of clinical management decisions such as infection control and
antibiotic use.
LAB-ON-A-CHIP (LOC) BASED MOLECULAR DIAGNOSTICS
Molecular diagnostic systems based on microfluidic technologies provide the means to automate and speed up molecular diagnostic assays. The quicker detection times are primarily due to the extremely low volumes involved, automation, and the low- overhead inbuilt cascading of the diagnostic process steps within a microfluidic device. Volumes in the nanoliter and microliter scale also reduce reagent consumption and cost. Lab-on-a-chip (LOC) devices are a common form of microfluidic device. LOC devices have MST structures within a MST layer for fluid processing integrated onto a single supporting substrate (usually silicon). Fabrication using the VLSI (very large scale integrated) lithographic techniques of the semiconductor industry keeps the unit cost of each LOC device very low. However, controlling fluid flow through the LOC device, adding reagents, controlling reaction conditions and so on necessitate bulky external plumbing and electronics. Connecting a LOC device to these external devices effectively restricts the use of LOC devices for molecular diagnostics to the laboratory setting. The cost of the external equipment and complexity of its operation precludes LOC-based molecular diagnostics as a practical option for point-of-care settings.
In view of the above, there is a need for a molecular diagnostic system based on a LOC device for use at point-of-care.
SUMMARY OF THE INVENTION
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Various aspects of the present invention are now described in the following numbered paragraphs.
GLE001.1 This aspect of the invention provides a micro fluidic device for analyzing a sample fluid, the microfluidic device comprising:
a sample inlet for receiving the sample;
a microsystems technology (MST) layer with functional sections for processing and analyzing the sample; and,
CMOS circuitry with digital memory for storing data and operational information to operatively control the functional sections during processing and analysis of the sample.
GLE001.2 Preferably, the microfluidic device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory relates to the reagent identities.
GLE001.3 Preferably, the data stored in the digital memory is a unique identifier for the microfluidic device.
GLE001.4 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
GLE001.5 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
GLE001.6 Preferably, the microfluidic device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
GLE001.7 Preferably, the microfluidic device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GLE001.8 Preferably, the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
GLE001.9 Preferably, each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and
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the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
GLE001.10 Preferably, the data stored in the digital memory includes hybridization data generated from the photosensor output.
GLE001.1 1 Preferably, the micro fluidic device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GLE001.12 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GLE001.13 Preferably, the CMOS circuitry has bond-pads and is configured for transmission of the hybridization data to an external device.
GLE001.14 Preferably, the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the bond-pads and store the patient data in the digital memory.
GLE001.15 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
GLE001.16 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
GLE001.17 Preferably, the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
GLE001.18 Preferably, the micro fluidic device also has a supporting substrate and a cap wherein the CMOS circuitry is between the supporting substrate and the MST layer, and the cap overlies the MST layer and defines the reagent reservoirs.
GLE001.19 Preferably, the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
GLE001.20 Preferably, the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
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The easily usable, mass-producible, and inexpensive microfluidic device with integral digital memory accepts an input fluid and processes it. The digital memory is used to store the data and control information required during the functioning of the device and the module incorporating the device. The digital memory being integral to the device, provides for an easily manufacturable, mass-producible, easily usable, and inexpensive microfluidic system with low component-count.
GLE002.1 This aspect of the invention provides a test module for analyzing a sample fluid, the test module comprising:
a receptacle for receiving the sample;
functional sections for processing and analyzing the sample; and,
digital memory for storing data and operational information to operatively control the functional sections during processing and analysis of the sample.
GLE002.2 Preferably, the test module also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory relates to the reagent identities.
GLE002.3 Preferably, the data stored in the digital memory is a unique identifier for the test module.
GLE002.4 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
GLE002.5 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
GLE002.6 Preferably, the test module also has CMOS circuitry and a temperature sensor wherein the CMOS circuitry incorporates the digital memory and uses the temperature sensor output for feedback control of the incubation section.
GLE002.7 Preferably, the test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GLE002.8 Preferably, the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
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GLE002.9 Preferably, each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
GLE002.10 Preferably, the data stored in the digital memory includes hybridization data generated from the photosensor output.
GLE002.1 1 Preferably, the test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GLE002.12 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GLE002.13 Preferably, the CMOS circuitry has a data interface for transmission of the hybridization data to an external device.
GLE002.14 Preferably, the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the data interface and store the patient data in the digital memory.
GLE002.15 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the control circuitry.
GLE002.16 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
GLE002.17 Preferably, the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
GLE002.18 Preferably, the test module also has a LOC device having a sample inlet in fluid communication with the receptacle, a supporting substrate, a microsystems technology (MST) layer, CMOS circuitry between the supporting substrate and the MST layer and a cap wherein the CMOS circuitry incorporates the digital memory, the MST layer incorporates the functional sections, and the cap overlies the MST layer and defines the reagent reservoirs.
GLE002.19 Preferably, the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that
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contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
GLE002.20 Preferably, the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
The easily usable, mass-producible, and inexpensive LOC device with integral digital memory accepts an input fluid and processes it. The digital memory is used to store the data and control information required during the functioning of the LOC device and the module incorporating the LOC device. The information stored on the memory includes the characteristics of the module incorporating this LOC device. The digital memory being integral to the device, provides for an easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
GLE003.1 This aspect of the invention provides a lab-on-a-chip (LOC) device for analyzing a sample fluid, the LOC device comprising:
a sample inlet for receiving the sample;
a microsystems technology (MST) layer with functional sections for processing and analyzing the sample; and,
CMOS circuitry with digital memory for storing epidemiological data, and configured to download epidemiological data updates from an external source.
GLE003.2 Preferably, the CMOS circuitry incorporates a universal serial bus (USB) device driver for operative control of a USB connection to the external source.
GLE003.3 Preferably, the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory relates to the reagent identities.
GLE003.4 Preferably, the data stored in the digital memory is a unique identifier for LOC device.
GLE003.5 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample.
GLE003.6 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an enzymatic reaction temperature.
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GLE003.7 Preferably, the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
GLE003.8 Preferably, the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GLE003.9 Preferably, each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
GLE003.10 Preferably, the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GLE003.1 1 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GLE003.12 Preferably, the CMOS circuitry has bond-pads for connection to the USB connection and transmission of hybridization data to an external device.
GLE003.13 Preferably, the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the USB connection and store the patient data in the digital memory.
GLE003.14 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
GLE003.15 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
GLE003.16 Preferably, the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
GLE003.17 Preferably, the LOC device also has a supporting substrate and a cap wherein the CMOS circuitry is between the supporting substrate and the MST layer, and the cap overlies the MST layer and defines the reagent reservoirs.
GLE003.18 Preferably, the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that
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contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
GLE003.19 Preferably, the PCR section has a thermal cycle time of less than 4 seconds.
GLE003.20 Preferably, the PCR section has a thermal cycle time between 0.45 seconds and 1.5 seconds.
The easily usable, mass-producible, and inexpensive LOC device with integral digital memory accepts an input fluid and processes it. The digital memory is used to store the data and control information required during the functioning of the LOC device and the module incorporating the LOC device. The information stored in the memory includes epidemiological updates available at the time, with the information being used for analytical and diagnostics purposes. This information provides for module's independence from outside support. The digital memory being integral to the device, provides for an easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
GLE004.1 This aspect of the invention provides a lab-on-a-chip (LOC) device for analyzing a sample fluid, the LOC device comprising:
a sample inlet for receiving the sample;
a microsystems technology (MST) layer with functional sections for processing and analyzing the sample; and,
CMOS circuitry with digital memory for storing genetic data, and configured to download genetic data updates from an external source.
GLE004.2 Preferably, the CMOS circuitry incorporates a universal serial bus
(USB) device driver for operative control of a USB connection to the external source.
GLE004.3 Preferably, the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory relates to the reagent identities.
GLE004.4 Preferably, the data stored in the digital memory is a unique identifier for LOC device.
GLE004.5 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the genetic material.
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GLE004.6 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an enzymatic reaction temperature.
GLE004.7 Preferably, the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
GLE004.8 Preferably, the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GLE004.9 Preferably, each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
GLE004.10 Preferably, the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GLE004.1 1 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GLE004.12 Preferably, the CMOS circuitry has bond-pads for transmission of hybridization data via the USB connection.
GLE004.13 Preferably, the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the bond-pads and store the patient data in the digital memory.
GLE004.14 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
GLE004.15 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
GLE004.16 Preferably, the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
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GLE004.17 Preferably, the LOC device also has a supporting substrate and a cap wherein the CMOS circuitry is between the supporting substrate and the MST layer, and the cap overlies the MST layer and defines the reagent reservoirs.
GLE004.18 Preferably, the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
GLE004.19 Preferably, the PCR section has a thermal cycle time of less than 4 seconds.
GLE004.20 Preferably, the PCR section has a thermal cycle time between 0.45 seconds and 1.5 seconds.
The easily usable, mass-producible, and inexpensive LOC device with integral digital memory accepts an input fluid and processes it. The digital memory is used to store the data and control information required during the functioning of the LOC device and the module incorporating the LOC device. The information stored in the memory includes genetic information updates available at the time, with the information being used for analytical and diagnostics purposes. This information provides for module's independence from outside support. The digital memory being integral to the device, provides for an easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
GLE005.1 This aspect of the invention provides a lab-on-a-chip (LOC) device for analyzing a sample fluid, the LOC device comprising:
a sample inlet for receiving the sample;
a microsystems technology (MST) layer with functional sections for processing and analyzing the sample; and,
CMOS circuitry with digital memory for storing data and operational information to operatively control the functional sections during processing and analysis of the sample; wherein,
the data is encrypted for secure communication with an external device.
GLE005.2 Preferably, the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory relates to the reagent identities.
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GLE005.3 Preferably, the data stored in the digital memory is a unique identifier for LOC device.
GLE005.4 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
GLE005.5 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
GLE005.6 Preferably, the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
GLE005.7 Preferably, the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GLE005.8 Preferably, the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
GLE005.9 Preferably, each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
GLE005.10 Preferably, the data stored in the digital memory includes hybridization data generated from the photosensor output.
GLE005.1 1 Preferably, the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GLE005.12 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GLE005.13 Preferably, the CMOS circuitry has bond-pads and is configured for transmission of the hybridization data to an external device.
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GLE005.14 Preferably, the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the bond-pads and store the patient data in the digital memory.
GLE005.15 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
GLE005.16 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
GLE005.17 Preferably, the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
GLE005.18 Preferably, the LOC device also has a supporting substrate and a cap wherein the CMOS circuitry is between the supporting substrate and the MST layer, and the cap overlies the MST layer and defines the reagent reservoirs.
GLE005.19 Preferably, the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
GLE005.20 Preferably, the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
The easily usable, mass-producible, and inexpensive LOC device with an integral digital memory accepts a diagnostic sample and processes it. The digital memory is used to store the data and control information required during the functioning of the LOC device and the diagnostic module incorporating the LOC device. The memory also securely stores patient test result information. The capability to store patient test result information makes it possible for the diagnostic module to perform a test utilizing only a minimal external power supply, and then in conjunction with a fully featured reader, analyze the patient test results at a later time. The secure storage of patient test result information would insure that the information would not be misused through illicit channels. The digital memory being integral to the device, provides for an easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
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GLE006.1 This aspect of the invention provides a test module for analyzing a sample fluid, the test module comprising:
a receptacle for receiving the sample;
functional sections for processing and analyzing the sample;
a supporting substrate; and,
CMOS circuitry on the supporting substrate for operative control of the functional sections during processing and analysis of the sample.
GLE006.2 Preferably, the test module also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the CMOS circuitry has digital memory for storing data relating to the reagent identities.
GLE006.3 Preferably, the data stored in the digital memory is a unique identifier for test module.
GLE006.4 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
GLE006.5 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
GLE006.6 Preferably, the test module also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
GLE006.7 Preferably, the test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GLE006.8 Preferably, the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
GLE006.9 Preferably, each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
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GLE006.10 Preferably, the data stored in the digital memory includes hybridization data generated from the photosensor output.
GLE006.1 1 Preferably, the test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GLE006.12 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GLE006.13 Preferably, the control circuitry has a data interface for transmission of the hybridization data to an external device.
GLE006.14 Preferably, the sample is drawn from a patient and the control circuitry is configured to download patient data via the data interface and store the patient data in the digital memory.
GLE006.15 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the control circuitry.
GLE006.16 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
GLE006.17 Preferably, the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
GLE006.18 Preferably, the test module also has a LOC device that incorporates the supporting substrate and the CMOS circuitry, and has a sample inlet in fluid communication with the receptacle, a microsystems technology (MST) layer that incorporates the functional sections and a cap wherein the cap overlies the MST layer and defines the reagent reservoirs.
GLE006.19 Preferably, the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
GLE006.20 Preferably, the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
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The easily usable, mass-producible, and inexpensive LOC device accepts a diagnostic sample and processes it, with the LOC device's on-chip electronics controlling all of the LOC device's functions. The control electronics being integral to the LOC device, makes the module utilizing the LOC device independent from specialized outside support and provides for the easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
GLE007.1 This aspect of the invention provides a lab-on-a-chip (LOC) device for installation in a test module for analyzing a sample fluid and communicating test results to an external device, the LOC device comprising:
a supporting substrate;
a sample inlet for receiving the sample;
a microsystems technology (MST) layer with functional sections for processing and analyzing the sample; and,
CMOS circuitry on the supporting substrate for operative control of a
communication interface in the test module for communication with the external device.
GLE007.2 Preferably, the CMOS circuitry incorporates a universal serial bus (USB) device driver for operative control of a USB plug in the test module for communication with an external device.
GLE007.3 Preferably, the CMOS circuitry is further configured for operative control of the functional sections during processing and analysis of the sample.
GLE007.4 Preferably, the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the CMOS circuitry has digital memory for storing data relating to the reagent identities.
GLE007.5 Preferably, the data stored in the digital memory is a unique identifier for LOC device.
GLE007.6 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
GLE007.7 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample
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and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
GLE007.8 Preferably, the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
GLE007.9 Preferably, the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GLE007.10 Preferably, the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
GLE007.1 1 Preferably, each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
GLE007.12 Preferably, the data stored in the digital memory includes hybridization data generated from the photosensor output.
GLE007.13 Preferably, the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GLE007.14 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GLE007.15 Preferably, the CMOS circuitry is configured for transmission of the hybridization data to an external device via the test module communications interface.
GLE007.16 Preferably, the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the communications interface and store the patient data in the digital memory.
GLE007.17 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the control circuitry.
GLE007.18 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
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GLE007.19 Preferably, the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
GLE007.20 Preferably, the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
The easily usable, mass-producible, and inexpensive LOC device accepts a diagnostic sample and processes it, with the LOC device's on-chip electronics controlling the data and command communications with the host. The electronics being integral to the LOC device, makes the module utilizing the LOC device independent from specialized outside support and provides for the easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
GLE008.1 This aspect of the invention provides a lab-on-a-chip (LOC) device for installation in a test module for analyzing a sample fluid and communicating test results to an external device, the LOC device comprising:
a supporting substrate;
a sample inlet for receiving the sample;
a microsystems technology (MST) layer with functional sections for processing and analyzing the sample; and,
CMOS circuitry on the supporting substrate, the CMOS circuitry having a universal serial bus (USB) device driver for operative control of a USB plug in the test module for communication with the external device.
GLE008.2 Preferably, the CMOS circuitry is further configured for operative control of the functional sections during processing and analysis of the sample.
GLE008.3 Preferably, the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the CMOS circuitry has digital memory for storing data relating to the reagent identities.
GLE008.4 Preferably, the data stored in the digital memory is a unique identifier for LOC device.
GLE008.5 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
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GLE008.6 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
GLE008.7 Preferably, the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
GLE008.8 Preferably, the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GLE008.9 Preferably, the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
GLE008.10 Preferably, each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
GLE008.1 1 Preferably, the data stored in the digital memory includes hybridization data generated from the photosensor output.
GLE008.12 Preferably, the digital memory includes random access memory
(RAM) and flash memory, the RAM being configured to store the hybridization data and the flash memory being configured to store program data to operate the functional sections and the probe identity data.
GLE008.13 Preferably, the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GLE008.14 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GLE008.15 Preferably, the USB device driver is configured for transmission of the hybridization data to an external device via the USB plug.
GLE008.16 Preferably, the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the USB plug and store the patient data in the digital memory.
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GLE008.17 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the control circuitry.
GLE008.18 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
GLE008.19 Preferably, the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
GLE008.20 Preferably, the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
The easily usable, mass-producible, and inexpensive LOC device with integral USB device controller accepts a diagnostic sample and processes it. The USB device controller being integral to the LOC device, makes the module utilizing the LOC device independent from specialized outside support and provides for the easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
GLE009.1 This aspect of the invention provides a lab-on-a-chip (LOC) device for analyzing a sample fluid, the LOC device comprising:
a supporting substrate;
a sample inlet for receiving the sample;
a microsystems technology (MST) layer with functional sections for processing and analyzing the sample; and,
CMOS circuitry between the supporting substrate and the MST layer, the CMOS circuitry having a controller to control operations performed by the functional sections during processing and analysis of the sample.
GLE009.2 Preferably, the CMOS circuitry has digital memory for storing data and operational information for use by the controller to control the functional sections during processing and analysis of the sample.
GLE009.3 Preferably, the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory is a unique identifier for LOC device, the unique identifier being associated with the reagent identities.
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GLE009.4 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
GLE009.5 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
GLE009.6 Preferably, the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
GLE009.7 Preferably, the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GLE009.8 Preferably, the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
GLE009.9 Preferably, each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
GLE009.10 Preferably, the data stored in the digital memory includes hybridization data generated from the photosensor output.
GLE009.1 1 Preferably, the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GLE009.12 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GLE009.13 Preferably, the CMOS circuitry has bond-pads and is configured for transmission of the hybridization data to an external device.
GLE009.14 Preferably, the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the bond-pads and store the patient data in the digital memory.
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GLE009.15 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
GLE009.16 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
GLE009.17 Preferably, the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
GLE009.18 Preferably, the LOC device also has a cap wherein the cap overlies the MST layer and defines the reagent reservoirs.
GLE009.19 Preferably, the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
GLE009.20 Preferably, the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
The easily usable, mass-producible, and inexpensive LOC device accepts a diagnostic sample and processes it, with the LOC device's integral controller controlling all of the LOC device's functions. The controller being integral to the LOC device, makes the module utilizing the LOC device independent from specialized outside support and provides for the easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
GLE010.1 This aspect of the invention provides a lab-on-a-chip (LOC) device for analyzing a sample fluid, the LOC device comprising:
a supporting substrate;
a sample inlet for receiving the sample;
a microsystems technology (MST) layer with functional sections for processing and analyzing the sample; and,
CMOS circuitry between the supporting substrate and the MST layer, the CMOS circuitry having digital memory for storing data and operational information to operatively control the functional sections during processing and analysis of the sample.
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GLE010.2 Preferably, the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory relates to the reagent identities.
GLE010.3 Preferably, the data stored in the digital memory is a unique identifier for LOC device.
GLE010.4 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
GLE010.5 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
GLE010.6 Preferably, the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
GLE010.7 Preferably, the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GLE010.8 Preferably, the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
GLE010.9 Preferably, each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
GLE010.10 Preferably, the data stored in the digital memory includes hybridization data generated from the photosensor output.
GLE010.1 1 Preferably, the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GLE010.12 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
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GLE010.13 Preferably, the CMOS circuitry has bond-pads and is configured for transmission of the hybridization data to an external device.
GLE010.14 Preferably, the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the bond-pads and store the patient data in the digital memory.
GLE010.15 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
GLEO 10.16 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
GLEO 10.17 Preferably, the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
GLEO 10.18 Preferably, the LOC device also has a cap wherein the cap overlies the MST layer and defines the reagent reservoirs.
GLEO 10.19 Preferably, the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
GLEO 10.20 Preferably, the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
The easily usable, mass-producible, and inexpensive LOC device accepts a diagnostic sample and processes it, with the LOC device's integral data RAM providing for intermediate data storage. The data RAM being integral to the LOC device, makes the module utilizing the LOC device independent from specialized outside support and provides for the easily manufacturable, mass-producible, easily usable, and inexpensive module with low component-count.
GLEO 1 1.1 This aspect of the invention provides a lab-on-a-chip (LOC) device for analyzing a sample fluid, the LOC device comprising:
a supporting substrate;
a sample inlet for receiving the sample;
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a microsystems technology (MST) layer with functional sections for processing and analyzing the sample; and,
CMOS circuitry between the supporting substrate and the MST layer, the CMOS circuitry having flash memory for storing program data to operate the functional sections during processing and analysis of the sample.
GLE01 1.2 Preferably, the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the flash memory also stores data relating to the reagent identities.
GLE01 1.3 Preferably, the data includes a unique identifier for LOC device. GLE01 1.4 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample, and the operational information stored in the digital memory relates to thermal cycle timing and duration.
GLE01 1.5 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
GLE01 1.6 Preferably, the LOC device also has a temperature sensor wherein the CMOS circuitry uses the temperature sensor output for feedback control of the incubation section.
GLE01 1.7 Preferably, the LOC device also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GLE01 1.8 Preferably, the flash memory stores probe identity data identifying the probe at each site within the array of probes.
GLE01 1.9 Preferably, each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to an input, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
GLE01 1.10 Preferably, the CMOS circuitry has random access memory (RAM) configured to store hybridization data generated from the photosensor output.
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GLE01 1.1 1 Preferably, the LOC device also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GLE01 1.12 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GLE01 1.13 Preferably, the CMOS circuitry has bond-pads and is configured for transmission of the hybridization data to an external device.
GLE01 1.14 Preferably, the sample is drawn from a patient and the CMOS circuitry is configured to download patient data via the bond-pads and store the patient data in the digital memory.
GLEO 1 1.15 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
GLEO 1 1.16 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
GLEO 1 1.17 Preferably, the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
GLEO 1 1.18 Preferably, the LOC device also has a cap wherein the cap overlies the MST layer and defines the reagent reservoirs.
GLEO 1 1.19 Preferably, the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
GLEO 1 1.20 Preferably, the PCR section is configured to complete a thermal cycle of the sample in less than 30 seconds.
The easily usable, mass-producible, and inexpensive microfluidic device with integral program and data flash memory accepts an input fluid and processes it. The flash memory is used to store the data and program required during the functioning of the device and the module incorporating the device. The flash memory being integral to the device, provides for an easily manufacturable, mass-producible, easily usable, and inexpensive microfluidic system with low component-count.
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GLE012.1 This aspect of the invention provides a test module for analyzing a sample fluid and communicating epidemiological data to a database, the test module comprising:
a receptacle for receiving the sample;
functional sections for processing and analyzing the sample;
a communication interface for communication with the epidemiological database; and,
a controller for operative control of the communication interface.
GLE012.2 Preferably, the test module also has a universal serial bus (USB) plug wherein the communication interface is a device driver for operative control of the USB plug to communicate with an external device.
GLE012.3 Preferably, the test module also has digital memory for storing epidemiological data.
GLE012.4 Preferably, the test module also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory includes the reagent identities.
GLE012.5 Preferably, the data stored in the digital memory includes a unique identifier for test module.
GLE012.6 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample.
GLE012.7 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an enzymatic reaction temperature.
GLE012.8 Preferably, the test module also has CMOS circuitry and a temperature sensor wherein the CMOS circuitry incorporates the digital memory and uses the temperature sensor output for feedback control of the incubation section.
GLE012.9 Preferably, the test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GLE012.10 Preferably, each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to excitation, and
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the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
GLE012.1 1 Preferably, the test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GLE012.12 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GLE012.13 Preferably, the CMOS circuitry is configured for communication of hybridization data with an external device.
GLE012.14 Preferably, the sample is drawn from a patient and the CMOS circuitry is configured to download and store the patient data in the digital memory.
GLE012.15 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
GLEO 12.16 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
GLEO 12.17 Preferably, the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
GLEO 12.18 Preferably, the test module also has a LOC device having a sample inlet in fluid communication with the receptacle, a supporting substrate, a microsystems technology (MST) layer, CMOS circuitry between the supporting substrate and the MST layer and a cap wherein the CMOS circuitry incorporates the digital memory, the communication interface, the controller, the MST layer incorporates the functional sections, and the cap overlies the MST layer and defines the reagent reservoirs.
GLEO 12.19 Preferably, the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
GLEO 12.20 Preferably, the PCR section has a thermal cycle time of less than 4 seconds.
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The easily usable, mass-producible, inexpensive, and portable diagnostic test module accepts a biochemical sample and processes and analyzes the sample, updating epidemiological databases based on the diagnostic results derived from the sample. The updating of epidemiological databases provides for improved science-base for the functioning of the diagnostic test modules and optimal higher- level responses to epidemiological situations. The diagnostic test module based automation of updating the databases would provide for massive quality and economic gains for health information systems.
GLE013.1 This aspect of the invention provides a test module for analyzing a sample fluid and communicating location data to an epidemiological database, the test module comprising:
a receptacle for receiving the sample;
functional sections for processing and analyzing the sample;
a communication interface for communication with the epidemiological database; and,
a controller for operative control of the communication interface; wherein, the controller is configured to associate location data with epidemiological data sent to the communication interface for communication with the epidemiological database.
GLE013.2 Preferably, the test module also has a universal serial bus (USB) plug wherein the communication interface is a USB device driver for operative control of the USB plug to communicate with an external device.
GLE013.3 Preferably, the controller is configured to automatically
communicate with the epidemiological database without user initiation.
GLE013.4 Preferably, the test module also has a user interface for inputting data to the controller for communication with the epidemiological database.
GLE013.5 Preferably, the test module also has digital memory for storing epidemiological data.
GLE013.6 Preferably, the test module also has a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory includes the reagent identities.
GLE013.7 Preferably, the data stored in the digital memory includes a unique identifier for test module.
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GLE013.8 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a polymerase chain reaction (PCR) section for amplifying nucleic acid sequences in the sample.
GLE013.9 Preferably, the test module also has CMOS circuitry and a temperature sensor wherein the CMOS circuitry incorporates the digital memory and uses the temperature sensor output for feedback control of the PCR section.
GLEO 13.10 Preferably, the test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GLEO 13.1 1 Preferably, each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to excitation, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
GLEO 13.12 Preferably, the test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GLEO 13.13 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GLEO 13.14 Preferably, the CMOS circuitry is configured for communication of hybridization data to an external device.
GLEO 13.15 Preferably, the sample is drawn from a patient and the CMOS circuitry is configured to download and store the patient data in the digital memory.
GLEO 13.16 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
GLEO 13.17 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
GLEO 13.18 Preferably, the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
GLEO 13.19 Preferably, the test module also has a LOC device having a sample inlet in fluid communication with the receptacle, a supporting substrate, a microsystems technology (MST) layer, CMOS circuitry between the supporting substrate and the MST
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layer and a cap wherein the CMOS circuitry incorporates the digital memory, the communication interface, the controller, the MST layer incorporates the functional sections, and the cap overlies the MST layer and defines the reagent reservoirs.
GLE013.20 Preferably, the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
The easily usable, mass-producible, inexpensive, and portable diagnostic test module accepts a biochemical sample and processes and analyzes the sample, updating epidemiological databases based on the diagnostic results derived from the sample and the test location data.
The updating of epidemiological databases with the diagnostics results and the location data provides for improved science-base for the functioning of the diagnostic test modules and optimal higher-level responses to epidemiological situations. The diagnostic test module based automation of updating the databases would provide for massive quality and economic gains for health information systems.
GLE014.1 This aspect of the invention provides a test module for analyzing a sample fluid and communicating data to a medical database, the test module comprising: a receptacle for receiving the sample;
functional sections for processing and analyzing the sample;
a communication interface for communication with the medical database; and, a controller for operative control of the communication interface.
GLE014.2 Preferably, the test module of claim 1 further comprising a universal serial bus (USB) plug wherein the communication interface is a USB device driver for operative control of the USB plug to communicate with an external device.
GLE014.3 Preferably, the test module of claim 2 further comprising digital memory wherein the medical database stores electronic health records (EH ), electronic medical records (EMR) and personal health records (PHR) and, the digital memory is configured for storing data relating to EHR, EMR and PHR.
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GLE014.4 Preferably, the test module of claim 3 further comprising a plurality of reagent reservoirs containing reagents for processing the sample wherein the data stored in the digital memory includes the reagent identities.
GLE014.5 Preferably, the data stored in the digital memory includes a unique identifier for test module.
GLE014.6 Preferably, the sample is a biological sample including cells of different sizes, and one of the functional sections is a polymerase chain reaction (PC ) section for amplifying nucleic acid sequences in the sample.
GLE014.7 Preferably, the test module of claim 6 further comprising CMOS circuitry and a temperature sensor wherein the CMOS circuitry incorporates the digital memory and uses the temperature sensor output for feedback control of the PCR section.
GLE014.8 Preferably, one of the functional sections is a dialysis section, the dialysis section being configured for separating cells larger than a predetermined threshold into a portion of the sample which is processed separately from the remainder of the sample containing only cells smaller than the predetermined threshold.
GLE014.9 Preferably, one of the functional sections is a lysis section, the lysis section being configured to release nucleic acid sequences within the cells smaller than the predetermined threshold.
GLE014.10 Preferably, the test module of claim 9 further comprising an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GLE014.1 1 Preferably, each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe-target hybrids being configured to emit photons in response to excitation, and the CMOS circuitry incorporates a photosensor for sensing the photons emitted by the probe-target hybrids.
GLE014.12 Preferably, the test module of claim 1 1 further comprising a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GLE014.13 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GLE014.14 Preferably, the CMOS circuitry is configured to communicate hybridization data to an external device.
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GLE014.15 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the CMOS circuitry.
GLEO 14.16 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
GLEO 14.17 Preferably, the meniscus anchor is an aperture and the heater has an annular shape and is positioned near the aperture periphery.
GLEO 14.18 Preferably, the test module of claim 5 further comprising a LOC device having a sample inlet in fluid communication with the receptacle, a supporting substrate, a microsystems technology (MST) layer, CMOS circuitry between the supporting substrate and the MST layer and a cap wherein the CMOS circuitry incorporates the digital memory, the communication interface, the controller, the MST layer incorporates the functional sections, and the cap overlies the MST layer and defines the reagent reservoirs.
GLEO 14.19 Preferably, the reagent reservoirs each have a surface tension valve with a meniscus anchor for pinning a meniscus to retain the reagent therein, such that contact with a flow of the sample fluid removes the meniscus and the reagent combines with the sample.
GLEO 14.20 Preferably, the PCR section has a thermal cycle time of less than 4 seconds.
The easily usable, mass-producible, inexpensive, and portable diagnostic test module accepts a biochemical sample and processes and analyzes the sample, updating patients' databases based on the diagnostic results derived from the sample.
The updating of patients' databases with the diagnostics results and the location data provides for improved provision of health care for the patients, automated maintenance of patient's medical records, improved science-base for the functioning of the diagnostic test modules, and optimal higher-level responses to epidemiological situations. The diagnostic test module based automation of updating the databases would provide for massive quality and economic gains for health information systems.
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GRE001.1 This aspect of the invention provides a micro flui die test module for analyzing a sample fluid and communicating test results to a mobile telephone, the microfluidic test module comprising:
an outer casing with a receptacle for receiving the sample;
functional sections for processing and analyzing the sample;
a communication interface for communication with the mobile telephone; and, a controller for operative control of the communication interface.
GRE001.2 Preferably, the communication interface is a universal serial bus (USB) device driver for operative control of a USB plug in the test module for communication with an external device.
GRE001.3 Preferably, the USB device driver is configured to draw power from the mobile telephone to power the controller and the functional sections.
GRE001.4 Preferably, the controller is further configured for operative control of the functional sections during processing and analysis of the sample.
GRE001.5 Preferably, the controller is configured to download data via the mobile telephone.
GRE001.6 Preferably, the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing data relating to the reagent identities.
GRE001.7 Preferably, the data stored in the digital memory is a unique identifier for the microfluidic test module.
GRE001.8 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a nucleic acid amplification section for amplifying nucleic acid sequences in the genetic material.
GRE001.9 Preferably, the nucleic acid amplification section is a polymerase chain reaction (PCR) section and the data stored in the digital memory includes thermal cycle times and cycle numbers.
GRE001.10 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
GRE001.11 Preferably, the microfluidic test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
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GRE001.12 Preferably, the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
GREOO 1.13 Preferably, the micro fluidic test module also has a photosensor wherein each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe- target hybrids being configured to emit photons in response to an input, and the photosensor is configured for sensing the photons emitted by the probe-target hybrids.
GREOO 1.14 Preferably, the data stored in the digital memory includes hybridization data generated from the photosensor output.
GREOO 1.15 Preferably, the micro fluidic test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GRE001.16 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GREOO 1.17 Preferably, the controller is configured for transmission of the hybridization data to the mobile telephone.
GREOO 1.18 Preferably, the sample is drawn from a patient and the controller is configured to download patient data via the mobile telephone and store the patient data in the digital memory.
GREOO 1.19 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the controller.
GREOO 1.20 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
The easily usable, mass-producible, inexpensive, compact, light, and portable Micro fluidic test module, with self-contained storage of required reagents, accepts a sample and processes and analyzes the sample material using the module's integral sensors, and provides the results electronically at its output port. The module's communications port is interfaced to a mobile phone/smart phone which provides the module with power, computing, communications, and user interface support. Mobile
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phones/smart phones are widely and inexpensively available, obviating the need for specialized, heavy, and expensive module support systems.
GRE002.1 This aspect of the invention provides a microfluidic test module for analyzing a sample fluid and communicating test results to a laptop computer, the microfluidic test module comprising:
an outer casing with a receptacle for receiving the sample;
functional sections for processing and analyzing the sample;
a communication interface for communication with the laptop computer; and, a controller for operative control of the communication interface.
GRE002.2 Preferably, the communication interface is a universal serial bus (USB) device driver for operative control of a USB plug in the test module for communication with an external device.
GRE002.3 Preferably, the USB device driver is configured to draw power from the laptop computer to power the controller and the functional sections.
GRE002.4 Preferably, the controller is further configured for operative control of the functional sections during processing and analysis of the sample.
GRE002.5 Preferably, the controller is configured to download data via the laptop computer.
GRE002.6 Preferably, the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing data relating to the reagent identities.
GRE002.7 Preferably, the data stored in the digital memory is a unique identifier for microfluidic test module.
GRE002.8 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a nucleic acid amplification section for amplifying nucleic acid sequences in the genetic material.
GRE002.9 Preferably, the nucleic acid amplification section is a polymerase chain reaction (PCR) section and the data stored in the digital memory includes thermal cycle times and cycle numbers.
GRE002.10 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample
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and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
GRE002.11 Preferably, the microfluidic test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GRE002.12 Preferably, the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
GRE002.13 Preferably, the microfluidic test module also has a photosensor wherein each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe- target hybrids being configured to emit photons in response to an input, and the photosensor is configured for sensing the photons emitted by the probe-target hybrids.
GRE002.14 Preferably, the data stored in the digital memory includes hybridization data generated from the photosensor output.
GRE002.15 Preferably, the microfluidic test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GRE002.16 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GRE002.17 Preferably, the controller is configured for transmission of the hybridization data to the laptop computer.
GRE002.18 Preferably, the sample is drawn from a patient and the controller is configured to download patient data via the laptop computer and store the patient data in the digital memory.
GRE002.19 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the controller.
GRE002.20 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
The easily usable, mass-producible, inexpensive, compact, light, and portable Microfluidic test module, with self-contained storage of required reagents, accepts a sample and processes and analyzes the sample material using the module's integral
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sensors, and provides the results electronically at its output port. The module's communications port is interfaced to a laptop/notebook which provides the module with power, computing, communications, and user interface support. Laptops/notebooks are widely and inexpensively available, obviating the need for specialized, heavy, and expensive module support systems.
GRE003.1 This aspect of the invention provides a micro flui die test module for analyzing a sample fluid and communicating test results to a dedicated reader purpose built for operating with the microfluidic test module, the microfluidic test module comprising: an outer casing with a receptacle for receiving the sample;
functional sections for processing and analyzing the sample;
a communication interface for communication with the dedicated reader; and, a controller for operative control of the communication interface.
GRE003.2 Preferably, the communication interface is a universal serial bus (USB) device driver for operative control of a USB plug in the test module for communication with an external device.
GRE003.3 Preferably, the USB device driver is configured to draw power from the dedicated reader to power the controller and the functional sections.
GRE003.4 Preferably, the controller is further configured for operative control of the functional sections during processing and analysis of the sample.
GRE003.5 Preferably, the controller is configured to download data via the dedicated reader.
GRE003.6 Preferably, the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing data relating to the reagent identities.
GRE003.7 Preferably, the data stored in the digital memory is a unique identifier for microfluidic test module.
GRE003.8 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a nucleic acid amplification section for amplifying nucleic acid sequences in the genetic material.
GRE003.9 Preferably, the nucleic acid amplification section is a polymerase chain reaction (PCR) section and the data stored in the digital memory includes thermal cycle times and cycle numbers.
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GRE003.10 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
GRE003.11 Preferably, the microfluidic test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GRE003.12 Preferably, the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
GRE003.13 Preferably, the microfluidic test module also has a photosensor wherein each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe- target hybrids being configured to emit photons in response to an input, and the photosensor is configured for sensing the photons emitted by the probe-target hybrids.
GRE003.14 Preferably, the data stored in the digital memory includes hybridization data generated from the photosensor output.
GRE003.15 Preferably, the microfluidic test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GRE003.16 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GRE003.17 Preferably, the controller is configured for transmission of the hybridization data to the dedicated reader.
GRE003.18 Preferably, the sample is drawn from a patient and the controller is configured to download patient data via the dedicated reader and store the patient data in the digital memory.
GRE003.19 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the controller.
GRE003.20 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
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The easily usable, mass-producible, inexpensive, compact, light, and portable Micro fluidic test module, with self-contained storage of required reagents, accepts a sample and processes and analyzes the sample material using the module's integral sensors, and provides the results electronically at its output port. The module's communications port is interfaced to an inexpensive and portable dedicated reader which provides the module with power, computing, communications, and user interface support. The dedicated reader obviates the need for heavy and expensive module support systems.
GRE004.1 This aspect of the invention provides a micro fluidic test module for analyzing a sample fluid and communicating test results to a desktop computer, the microfluidic test module comprising:
an outer casing with a receptacle for receiving the sample;
functional sections for processing and analyzing the sample;
a communication interface for communication with the desktop computer; and, a controller for operative control of the communication interface.
GRE004.2 Preferably, the communication interface is a universal serial bus (USB) device driver for operative control of a USB plug in the test module for communication with an external device.
GRE004.3 Preferably, the USB device driver is configured to draw power from the desktop computer to power the controller and the functional sections.
GRE004.4 Preferably, the controller is further configured for operative control of the functional sections during processing and analysis of the sample.
GRE004.5 Preferably, the controller is configured to download data via the desktop computer.
GRE004.6 Preferably, the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing data relating to the reagent identities.
GRE004.7 Preferably, the data stored in the digital memory is a unique identifier for microfluidic test module.
GRE004.8 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a nucleic acid amplification section for amplifying nucleic acid sequences in the genetic material.
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GRE004.9 Preferably, the nucleic acid amplification section is a polymerase chain reaction (PCR) section and the data stored in the digital memory includes thermal cycle times and cycle numbers.
GRE004.10 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
GRE004.11 Preferably, the micro fluidic test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GRE004.12 Preferably, the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
GRE004.13 Preferably, the micro fluidic test module also has a photosensor wherein each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe- target hybrids being configured to emit photons in response to an input, and the photosensor is configured for sensing the photons emitted by the probe-target hybrids.
GRE004.14 Preferably, the data stored in the digital memory includes hybridization data generated from the photosensor output.
GRE004.15 Preferably, the micro fluidic test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GRE004.16 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GRE004.17 Preferably, the controller is configured for transmission of the hybridization data to the desktop computer.
GRE004.18 Preferably, the sample is drawn from a patient and the controller is configured to download patient data via the desktop computer and store the patient data in the digital memory.
GRE004.19 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the controller.
GRE004.20 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the
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liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
The easily usable, mass-producible, inexpensive, compact, light, and portable Micro fluidic test module, with self-contained storage of required reagents, accepts a sample and processes and analyzes the sample material using the module's integral sensors, and provides the results electronically at its output port. The module's communications port is interfaced to a desktop PC which provides the module with power, computing, communications, and user interface support. Desktop PCs are widely available and are inexpensive, obviating the need for specialized, heavy, and expensive module support systems.
GRE005.1 This aspect of the invention provides a micro fluidic test module for analyzing a sample fluid and communicating test results to an ebook reader, the micro fluidic test module comprising:
an outer casing with a receptacle for receiving the sample;
functional sections for processing and analyzing the sample;
a communication interface for communication with the ebook reader; and, a controller for operative control of the communication interface.
GRE005.2 Preferably, the communication interface is a universal serial bus
(USB) device driver for operative control of a USB plug in the test module for communication with an external device.
GRE005.3 Preferably, the USB device driver is configured to draw power from the ebook reader to power the controller and the functional sections.
GRE005.4 Preferably, the controller is further configured for operative control of the functional sections during processing and analysis of the sample.
GRE005.5 Preferably, the controller is configured to download data via the ebook reader.
GRE005.6 Preferably, the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing data relating to the reagent identities.
GRE005.7 Preferably, the data stored in the digital memory is a unique identifier for microfluidic test module.
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GRE005.8 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a nucleic acid amplification section for amplifying nucleic acid sequences in the genetic material.
GRE005.9 Preferably, the nucleic acid amplification section is a polymerase chain reaction (PCR) section and the data stored in the digital memory includes thermal cycle times and cycle numbers.
GRE005.10 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
GRE005.11 Preferably, the micro fluidic test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GRE005.12 Preferably, the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
GRE005.13 Preferably, the microfluidic test module also has a photosensor wherein each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe- target hybrids being configured to emit photons in response to an input, and the photosensor is configured for sensing the photons emitted by the probe-target hybrids.
GRE005.14 Preferably, the data stored in the digital memory includes hybridization data generated from the photosensor output.
GRE005.15 Preferably, the microfluidic test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GRE005.16 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GRE005.17 Preferably, the controller is configured for transmission of the hybridization data to the ebook reader.
GRE005.18 Preferably, the sample is drawn from a patient and the controller is configured to download patient data via the ebook reader and store the patient data in the digital memory.
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GRE005.19 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the controller.
GRE005.20 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
The easily usable, mass-producible, inexpensive, compact, light, and portable Micro fluidic test module, with self-contained storage of required reagents, accepts a sample and processes and analyzes the sample material using the module's integral sensors, and provides the results electronically at its output port. The module's communications port is interfaced to a ebook reader which provides the module with power, computing, communications, and user interface support. Ebook readers are widely and inexpensively available, obviating the need for specialized, heavy, and expensive module support systems.
GRE007.1 This aspect of the invention provides a microfluidic test module for analyzing a sample fluid and communicating test results to a tablet computer, the microfluidic test module comprising:
an outer casing with a receptacle for receiving the sample;
functional sections for processing and analyzing the sample;
a communication interface for communication with the tablet computer; and, a controller for operative control of the communication interface.
GRE007.2 Preferably, the communication interface is a universal serial bus
(USB) device driver for operative control of a USB plug in the test module for communication with an external device.
GRE007.3 Preferably, the USB device driver is configured to draw power from the tablet computer to power the controller and the functional sections.
GRE007.4 Preferably, the controller is further configured for operative control of the functional sections during processing and analysis of the sample.
GRE007.5 Preferably, the controller is configured to download data via the tablet computer.
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GRE007.6 Preferably, the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing data relating to the reagent identities.
GRE007.7 Preferably, the data stored in the digital memory is a unique identifier for microfluidic test module.
GRE007.8 Preferably, the sample is a biological sample containing genetic material and one of the functional sections is a nucleic acid amplification section for amplifying nucleic acid sequences in the genetic material.
GRE007.9 Preferably, the nucleic acid amplification section is a polymerase chain reaction (PCR) section and the data stored in the digital memory includes thermal cycle times and cycle numbers.
GRE007.10 Preferably, the functional sections include an incubation section upstream of the PCR section and one of the reagent reservoirs is a restriction enzyme reservoir, the incubation section having a heater for maintaining a mixture of the sample and restriction enzymes at an incubation temperature during restriction digestion of the nucleic acid sequences.
GRE007.11 Preferably, the microfluidic test module also has an array of probes for hybridization with target nucleic acid sequences in the amplicon from the PCR section.
GRE007.12 Preferably, the data stored in the digital memory includes probe identity data identifying the probe at each site within the array of probes.
GRE007.13 Preferably, the microfluidic test module also has a photosensor wherein each of the probes are configured to form a probe-target hybrid with a complementary target nucleic acid sequence contained in the amplicon, each of the probe- target hybrids being configured to emit photons in response to an input, and the photosensor is configured for sensing the photons emitted by the probe-target hybrids.
GRE007.14 Preferably, the data stored in the digital memory includes hybridization data generated from the photosensor output.
GRE007.15 Preferably, the microfluidic test module also has a hybridization chamber array for containing the probes such that the probes within each hybridization chamber are configured to hybridize with one of the target nucleic acid sequences.
GRE007.16 Preferably, the photosensor is an array of photodiodes positioned in registration with the hybridization chambers.
GRE007.17 Preferably, the controller is configured for transmission of the hybridization data to the tablet computer.
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GRE007.18 Preferably, the sample is drawn from a patient and the controller is configured to download patient data via the tablet computer and store the patient data in the digital memory.
GRE007.19 Preferably, the PCR section has an active valve for retaining liquid in the PCR section during thermal cycling and allowing flow to the hybridization chambers in response to an activation signal from the controller.
GRE007.20 Preferably, the active valve is a boiling-initiated valve with a meniscus anchor configured to pin a meniscus that arrests capillary driven flow of the liquid, and a heater for boiling the liquid to unpin the meniscus from the meniscus anchor such that capillary driven flow resumes.
The easily usable, mass-producible, inexpensive, compact, light, and portable Micro fluidic test module, with self-contained storage of required reagents, accepts a sample and processes and analyzes the sample material using the module's integral sensors, and provides the results electronically at its output port. The module's communications port is interfaced to a tablet computer which provides the module with power, computing, communications, and user interface support. Tablet computers are widely and inexpensively available, obviating the need for specialized, heavy, and expensive module support systems.
GMV001.1 This aspect of the invention provides a reagent microvial for a reagent dispensing apparatus, the microvial comprising:
a container for holding a volume of reagent;
a digital memory for data relating to the reagent;
a droplet generator for ejecting droplets of the reagent from the container; and, electrical contacts for connection to a control processor in the reagent dispensing apparatus to receive drive pulses for the droplet generator and provide the data to the control processor.
GMV001.2 Preferably, the container holds between 282 microliters and 400 microliters of reagent.
GMV001.3 Preferably, the droplet generator is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
GMV001.4 Preferably, the droplet generator has a piezo-electric actuator.
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GMV001.5 Preferably, the data includes an identity transmitted to the control processor.
GMV001.6 Preferably, the identity is a unique identity distinguishing the microvial from all other microvials.
GMV001.7 Preferably, the data is encrypted.
The reagent microvial with nonvolatile digital memory is used to receive a reagent, store it, and dispense it under digital control using a droplet generator. The droplet generator being part of the microvial provides for a self-contained and volumetrically and positionally precise reagent dispensing technique, simplifying the complexity, increasing the reliability, and reducing the cost of the automated manufacturing environment utilizing the microvial.
The digital memory is used to store the information required during the functioning of the device in an automated manufacturing environment. The digital memory being part of the microvial, provides for an easily usable, reliable, safe, secure, and inexpensive approach to reagent dispensing in an automated manufacturing environment.
GMV002.1 This aspect of the invention provides a reagent microvial for a reagent dispensing apparatus, the microvial comprising:
a container for holding a volume of reagent;
an integrated circuit with digital memory for data relating to an identity of the microvial; and,
electrical contacts for connection to a control processor in the reagent dispensing apparatus to provide the data to the control processor for comparison with a list of authentic microvial identities.
GMV002.2 Preferably, the microvial also has a droplet generator for ejecting droplets of the reagent from the container.
GMV002.3 Preferably, the container holds between 282 microliters and 400 microliters of reagent.
GMV002.4 Preferably, the droplet generator is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
GMV002.5 Preferably, the droplet generator has a piezo-electric actuator.
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GMV002.6 Preferably, the identity is a unique identity distinguishing the microvial from all other microvials.
GMV002.7 Preferably, the data is encrypted. The reagent microvial with authentication integrated circuit is used to receive a reagent, store it, and dispense it under digital control using a droplet generator. The droplet generator being part of the microvial provides for a self-contained and volumetrically and positionally precise reagent dispensing technique, simplifying the complexity, increasing the reliability, and reducing the cost of the automated manufacturing environment utilizing the microvial.
The authentication integrated circuit is used to store the microvial authentication information used during the functioning of the device in an automated manufacturing environment. The authentication integrated circuit being part of the microvial, provides for an easily usable, reliable, safe, secure, and inexpensive approach to reagent dispensing in an automated manufacturing environment.
GMV003.1 This aspect of the invention provides a reagent microvial for a reagent dispensing apparatus, the microvial comprising:
a container for holding a volume of reagent;
an integrated circuit with digital memory storing reagent data relating to specifications characterizing the reagent; and,
electrical contacts for connection to a control processor in the reagent dispensing apparatus to provide the data to the control processor for download into devices supplied with the reagent.
GMV003.2 Preferably, the microvial also has a droplet generator for ejecting droplets of the reagent from the container.
GMV003.3 Preferably, the container holds between 282 microliters and 400 microliters of reagent.
GMV003.4 Preferably, the droplet generator is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
GMV003.5 Preferably, the droplet generator has a piezo-electric actuator.
GMV003.6 Preferably, the identity is a unique identity distinguishing the microvial from all other microvials.
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GMV003.7 Preferably, the data is encrypted.
The reagent microvial with nonvolatile digital memory is used to receive a reagent, store it, and dispense it under digital control using a droplet generator. The droplet generator being part of the microvial provides for a self-contained and volumetrically and positionally precise reagent dispensing technique, simplifying the complexity, increasing the reliability, and reducing the cost of the automated manufacturing environment utilizing the microvial. The digital memory is used to store the information required during the functioning of the device in an automated manufacturing environment. The information stored in the memory includes the reagent specification data written into the memory by segments of the automated manufacturing environment. This information gets read from this memory an utilized as required by other segments of the automated manufacturing environment. The digital memory being part of the microvial, provides for an easily usable, reliable, safe, secure, and inexpensive approach to reagent dispensing in an automated manufacturing environment.
GMV004.1 This aspect of the invention provides an oligonucleotide microvial for an oligonucleotide dispensing apparatus, the microvial comprising:
a container for holding a volume of reagent;
an integrated circuit with digital memory storing oligonucleotide data relating to specifications characterizing the oligonucleotides; and,
electrical contacts for connection to a control processor in the reagent dispensing apparatus to provide the data to the control processor for download into devices supplied with the oligonucleotides.
GMV004.2 Preferably, the microvial also has a droplet generator for ejecting droplets of the oligonucleotides from the container.
GMV004.3 Preferably, the container holds between 282 microliters and 400 microliters of reagent.
GMV004.4 Preferably, the droplet generator is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
GMV004.5 Preferably, the droplet generator has a piezo-electric actuator.
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GMV004.6 Preferably, the identity is a unique identity distinguishing the microvial from all other microvials.
GMV004.7 Preferably, the data is encrypted. The oligonucleotide microvial with nonvolatile digital memory is used to receive a reagent, store it, and dispense it under digital control using a droplet generator. The droplet generator being part of the microvial provides for a self-contained and volumetrically and positionally precise oligonucleotide dispensing technique, simplifying the complexity, increasing the reliability, and reducing the cost of the automated manufacturing environment utilizing the microvial.
The digital memory is used to store the information required during the functioning of the device in an automated manufacturing environment. The information stored in the memory includes the oligonucleotide specification data written into the memory by segments of the automated manufacturing environment. This information gets read from this memory an utilized as required by other segments of the automated manufacturing environment. The digital memory being part of the microvial, provides for an easily usable, reliable, safe, secure, and inexpensive approach to reagent dispensing in an automated manufacturing environment.
GRD001.1 This aspect of the invention provides a reagent dispensing apparatus for loading reagents into a microfluidic device having a digital memory for data related to the reagents loaded into the microfluidic device, the reagent dispensing apparatus comprising:
a plurality of reagent vials each of the vials having an integrated circuit with memory storing data regarding the reagent in the vial, and a droplet dispenser;
a mounting surface for detachably mounting the microfluidic device for movement relative to the vials; and,
a control processor for operative control of the vials and the mounting surface; wherein,
the control processor is configured to activate the droplet dispenser of the vial selected, move the vial into registration with the microfluidic device and download the data from the integrated circuit to the digital memory of the microfluidic device.
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GRD001.2 Preferably, the reagent dispensing apparatus also has a camera for optical feedback of the registration between the vial selected by the control processor and the microfluidic device.
GRD001.3 Preferably, the vial is a microvial for holding between 282 microliters and 400 microliters.
GRD001.4 Preferably, the integrated circuit for each of the microvials has a unique identifier for identifying each of the microvials individually, the unique identifier being transmitted to the control processor and the digital memory of the microfluidic device.
GRD001.5 Preferably, each of the microvials has electrical contacts for receiving activation pulses for the droplet dispenser and allowing the control processor to interrogate the integrated circuit.
GRD001.6 Preferably, the reagent dispensing apparatus also has a rack wherein the microvials are detachably mounted to the rack for mechanical and electronic control of the microvials.
GRD001.7 Preferably, the mounting surface is a stage configured for movement along two orthogonal axes, the rack extending parallel to one if the orthogonal axes.
GRD001.8 Preferably, the microfluidic device is a lab-on-a-chip (LOC) device.
GRD001.9 Preferably, the droplet dispenser has a piezo-electric actuator.
GRD001.10 Preferably, the droplet dispenser is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
GRD001.1 1 Preferably, the reagent dispensing apparatus also has facilities configured for applying a seal to the LOC device to close a plurality of reservoirs in which the reagents have been loaded.
GRD001.12 Preferably, the LOC has a polymerase chain reaction (PCR) section and the list of reagents has one or more of:
water;
polymerase;
primers;
buffer;
anticoagulant;
deoxyribonucleoside triphosphates (dNTPs);
lysis reagent;
ligase and linkers; and,
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restriction enzymes.
The reagent dispensing apparatus is used, as part of a cost-effective automated mass-manufacturing environment, to dispense reagents contained in reagent microvials into the reagent reservoirs of microfluidic devices. The data automation provided by the reagent dispensing apparatus includes automated computer-controlled dispensing of the reagents into the reagent reservoirs of the microfluidic devices, checking the reagent data stored in the memory of the microvials against the list of specifications for the reagents that have to be loaded in the microfluidic device, and storage of the reagent data into the memory of the microfluidic device.
The reagent dispensing apparatus provides for an automated and volumetrically and positionally precise reagent dispensing technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
The data automation provided by the reagent dispensing apparatus provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GRD002.1 This aspect of the invention provides a reagent dispensing apparatus for loading reagents into a microfluidic device having a digital memory for data related to the reagents loaded into the microfluidic device, the reagent dispensing apparatus comprising:
a plurality of reagent vials each of the vials having an integrated circuit with memory storing data regarding the reagent in the vial, and a droplet dispenser;
a mounting surface for detachably mounting the microfluidic device for movement relative to the vials; and,
a control processor for operative control of the vials and the mounting surface; wherein,
the control processor is configured to automatically interrogate each of the integrated circuits to collect and store the data regarding the reagents in each of the vials.
GRD002.2 Preferably, the control processor is configured to automatically activate the droplet dispenser of the vial selected, move the vial into registration with the
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microfluidic device and download information from the integrated circuit to the digital memory of the microfluidic device.
GRD002.3 Preferably, the reagent dispensing apparatus also has a camera for optical feedback of the registration between the vial selected by the control processor and the microfluidic device.
GRD002.4 Preferably, the vial is a microvial for holding between 282 microliters and 400 microliters.
GRD002.5 Preferably, the integrated circuit for each of the microvials has a unique identifier for identifying each of the microvials individually, the unique identifier being transmitted to the control processor and the digital memory of the microfluidic device.
GRD002.6 Preferably, each of the microvials has electrical contacts for receiving activation pulses for the droplet dispenser and allowing the control processor to interrogate the integrated circuit.
GRD002.7 Preferably, the reagent dispensing apparatus also has a rack wherein the microvials are detachably mounted to the rack for mechanical and electronic control of the microvials.
GRD002.8 Preferably, the mounting surface is a stage configured for movement along two orthogonal axes, the rack extending parallel to one if the orthogonal axes.
GRD002.9 Preferably, the microfluidic device is a lab-on-a-chip (LOC) device.
GRD002.10 Preferably, the droplet dispenser has a piezo-electric actuator.
GRD002.1 1 Preferably, the droplet dispenser is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
GRD002.12 Preferably, the LOC has a polymerase chain reaction (PCR) section and the list of reagents has one or more of:
water;
polymerase;
primers;
buffer;
anticoagulant;
deoxyribonucleoside triphosphates (dNTPs);
lysis reagent;
ligase and linkers; and,
restriction enzymes.
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The reagent dispensing apparatus is used, as part of a cost-effective automated mass-manufacturing environment, to dispense reagents contained in reagent microvials into the reagent reservoirs of microfluidic devices. The data automation provided by the reagent dispensing apparatus includes automated computer-controlled dispensing of the reagents into the reagent reservoirs of the microfluidic devices, checking the reagent data stored in the memory of the microvials against the list of specifications for the reagents that have to be loaded in the microfluidic device, and storage of the reagent data into the memory of the microfluidic device and in the reagent dispensing apparatus's computer memory.
The reagent dispensing apparatus provides for an automated and volumetrically and positionally precise reagent dispensing technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
The data automation provided by the reagent dispensing apparatus provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GRD003.1 This aspect of the invention provides a reagent dispensing apparatus for loading reagents into a fixed array of microfluidic devices, each microfluidic device having a digital memory for data related to the reagents loaded into the microfluidic device, the reagent dispensing apparatus comprising:
a plurality of reagent vials each of the vials having an integrated circuit with memory storing data regarding the reagent in the vial, and a droplet dispenser;
a mounting surface for detachably mounting the fixed array of microfluidic devices for movement relative to the vials; and,
a control processor for operative control of the vials and the mounting surface; wherein,
the control processor is configured to activate the droplet dispenser of the vial selected, move the vial into registration within one or more of the microfluidic devices within the fixed array and download the data from the integrated circuit to the digital memory of the one or more microfluidic devices.
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GRD003.2 Preferably, the fixed array of microfluidic devices is an array of lab- on-a-chip (LOC) devices mounted on a separable PCB (printed circuit board) wafer.
GRD003.3 Preferably, the reagent dispensing apparatus also has a camera for optical feedback of the registration between the vial selected by the control processor and the LOC device.
GRD003.4 Preferably, the vial is a microvial for holding between 282 microliters and 400 microliters.
GRD003.5 Preferably, the integrated circuit for each of the microvials has a unique identifier for identifying each of the microvials individually, the unique identifier being transmitted to the control processor and the digital memory of the LOC device.
GRD003.6 Preferably, each of the microvials has electrical contacts for receiving activation pulses for the droplet dispenser and allowing the control processor to interrogate the integrated circuit.
GRD003.7 Preferably, the reagent dispensing apparatus also has a rack wherein the microvials are detachably mounted to the rack for mechanical and electronic control of the microvials.
GRD003.8 Preferably, the mounting surface is a stage configured for movement along two orthogonal axes, the rack extending parallel to one if the orthogonal axes.
GRD003.9 Preferably, the droplet dispenser has a piezo-electric actuator.
GRD003.10 Preferably, the droplet dispenser is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
GRD003.1 1 Preferably, the reagent dispensing apparatus also has facilities configured for applying a seal to the LOC device to close a plurality of reservoirs in which the reagents have been loaded.
GRD003.12 Preferably, the LOC has a polymerase chain reaction (PCR) section and the list of reagents has one or more of:
water;
polymerase;
primers;
buffer;
anticoagulant;
deoxyribonucleoside triphosphates (dNTPs);
lysis reagent;
ligase and linkers; and,
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restriction enzymes.
GRD003.13 Preferably, the control processor is configured to automatically interrogate each of the integrated circuits to collect and store the data regarding the reagents in each of the vials.
The reagent dispensing apparatus is used, as part of a cost-effective automated mass-manufacturing environment, to dispense reagents contained in reagent microvials into the reagent reservoirs of arrays of micro fluidic devices mounted on PCB wafers. The data automation provided by the reagent dispensing apparatus includes automated computer-controlled dispensing of the reagents into the reagent reservoirs of the micro fluidic devices, checking the reagent data stored in the memory of the microvials against the list of specifications for the reagents that have to be loaded in the microfluidic device, and storage of the reagent data into the memory of the microfluidic device. The reagent dispensing apparatus provides for an automated and volumetrically and positionally precise reagent dispensing technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The data automation provided by the reagent dispensing apparatus provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
Dispensing of the reagents into the arrays of microfluidic devices mounted on PCB wafers speeds up and reduces the cost of the loading process, and by loading the reagents into the microfluidic devices after mounting the devices on the PCB wafer and soldering them, improves the chemical and physical integrity of the reagents.
GRD004.1 This aspect of the invention provides a reagent dispensing apparatus for loading reagents into a silicon wafer on which an array of lab-on-a-chip (LOC) devices are fabricated, each LOC device having a digital memory for data related to the reagents loaded into the LOC device, the reagent dispensing apparatus comprising:
a plurality of reagent vials each of the vials having an integrated circuit with memory storing data regarding the reagent in the vial, and a droplet dispenser;
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a mounting surface for detachably mounting the silicon wafer for movement relative to the vials; and,
a control processor for operative control of the vials and the mounting surface; wherein,
the control processor is configured to activate the droplet dispenser of the vial selected, move the vial into registration within one or more of the LOC devices on the silicon wafer and download the data from the integrated circuit to the digital memory of the one or more LOC devices.
GRD004.2 Preferably, the silicon wafer is partially sawn in preparation for tessellation into individually separate LOC devices.
GRD004.3 Preferably, the reagent dispensing apparatus also has a camera for optical feedback of the registration between the vial selected by the control processor and the LOC device.
GRD004.4 Preferably, the vial is a microvial for holding between 282 microliters and 400 microliters.
GRD004.5 Preferably, the integrated circuit for each of the microvials has a unique identifier for identifying each of the microvials individually, the unique identifier being transmitted to the control processor and the digital memory of the LOC device.
GRD004.6 Preferably, each of the microvials has electrical contacts for receiving activation pulses for the droplet dispenser and allowing the control processor to interrogate the integrated circuit.
GRD004.7 Preferably, the reagent dispensing apparatus also has a rack wherein the microvials are detachably mounted to the rack for mechanical and electronic control of the microvials.
GRD004.8 Preferably, the mounting surface is a stage configured for movement along two orthogonal axes, the rack extending parallel to one if the orthogonal axes.
GRD004.9 Preferably, the droplet dispenser has a piezo-electric actuator.
GRD004.10 Preferably, the droplet dispenser is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
GRD004.1 1 Preferably, the reagent dispensing apparatus also has facilities configured for applying a seal to the LOC device to close a plurality of reservoirs in which the reagents have been loaded.
GRD004.12 Preferably, the LOC has a polymerase chain reaction (PCR) section and the list of reagents has one or more of:
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water;
polymerase;
primers;
buffer;
anticoagulant;
deoxyribonucleoside triphosphates (dNTPs);
lysis reagent;
ligase and linkers; and,
restriction enzymes.
GRD004.13 Preferably, the control processor is configured to automatically interrogate each of the integrated circuits to collect and store the data regarding the reagents in each of the vials.
The reagent dispensing apparatus is used, as part of a cost-effective automated mass-manufacturing environment, to dispense reagents contained in reagent microvials into the reagent reservoirs of micro fluidic devices on partial-depth sawn silicon wafers. The data automation provided by the reagent dispensing apparatus includes automated computer-controlled dispensing of the reagents into the reagent reservoirs of the micro fluidic devices, checking the reagent data stored in the memory of the microvials against the list of specifications for the reagents that have to be loaded in the microfluidic device, and storage of the reagent data into the memory of the microfluidic device.
The reagent dispensing apparatus provides for an automated and volumetrically and positionally precise reagent dispensing technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
The data automation provided by the reagent dispensing apparatus provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
Dispensing of the reagents into microfluidic devices on partial-depth sawn silicon wafers speeds up the process of loading and reduces its cost.
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GPD001.1 This aspect of the invention provides an oligonucleotide spotting device for contactless spotting of oligonucleotide probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the oligonucleotide spotting device comprising:
a supporting substrate;
an array of reservoirs for containing the oligonucleotide probes suspended in a liquid; and,
an array of ejectors, each of the ejectors being configured for fluid communication with a corresponding one of the reservoirs respectively; wherein,
the ejectors are configured to eject droplets containing the oligonucleotide probes from the corresponding reservoir onto the surface.
GPD001.2 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPD001.3 Preferably, the ejector has a chamber for containing the liquid from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GPD001.4 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPD001.5 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
GPD001.6 Preferably, the ejectors are configured to eject droplets having a volume less than 25 picoliters.
GPD001.7 Preferably, the ejectors are configured to eject droplets having a volume less than 6 picoliters.
GPD001.8 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
GPD001.9 Preferably, the actuators in each of the ejectors are configured to actuate individually.
GPD001.10 Preferably, each of the ejectors has a plurality of inlet channels extending from the reservoir to the chamber.
GPD001.11 Preferably, the oligonucleotide spotting device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond- pads for connection to a microprocessor controller operatively controlling relative
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movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
GPD001.12 Preferably, the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
GPD001.13 Preferably, the supporting substrate has a reservoir side and an ejector side opposite the reservoir side wherein the array of reservoirs is formed in the reservoir side and the array of ejectors is formed on the ejector side.
GPD001.14 Preferably, the CMOS circuitry is between the array of reservoirs and the array of ejectors.
GPD001.15 Preferably, the array of ejectors is configured to eject droplets containing the oligonucleotide probes onto the surface with a density greater than 1 droplet per square millimeter.
GPD001.16 Preferably, the array of ejectors is configured to eject droplets containing the oligonucleotide probes onto the surface with a density greater than 8 droplets per square millimeter.
GPD001.17 Preferably, the array of ejectors is configured to eject droplets containing the oligonucleotide probes onto the surface with a density greater than 60 droplets per square millimeter.
GPD001.18 Preferably, the array of ejectors is configured to eject droplets containing the oligonucleotide probes onto the surface with a density between 500 droplets per square millimeter and 1500 droplets per square millimeter.
GPD001.19 Preferably, the array of ejectors is configured to eject droplets containing the oligonucleotide probes onto the surface at a rate greater than 100 droplets per second.
GPD001.20 Preferably, the array of ejectors is configured to eject droplets containing the oligonucleotide probes onto the surface at a rate greater than 1,400 droplets per second.
The mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted. The data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides onto the surface being spotted, receiving the specifications of the oligonucleotides stored in
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its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment. The oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
The data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GPD003.1 This aspect of the invention provides a spotting device for contactless spotting of a lab-on-a-chip (LOC) device with oligonucleotide probes, the LOC device having an array of hybridization chambers for receiving the oligonucleotide probes, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample and the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, the spotting device comprising:
an array of reservoirs for containing the oligonucleotide probes suspended in a liquid; and,
an array of ejectors, each of the ejectors being configured for fluid communication with a corresponding one of the reservoirs respectively such that the ejectors eject droplets containing the oligonucleotide probes from the corresponding reservoir into one of the hybridization chambers; wherein,
the array of reservoirs is configured to contain the complete assay of
oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
GPD003.2 Preferably, the array of reservoirs has more than 1000 reservoirs.
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GPD003.3 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPD003.4 Preferably, the ejector has a chamber for containing the liquid from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GPD003.5 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPD003.6 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
GPD003.7 Preferably, the ejectors are configured to eject droplets having a volume less than 25 picoliters.
GPD003.8 Preferably, the ejectors are configured to eject droplets having a volume less than 6 picoliters.
GPD003.9 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
GPD003.10 Preferably, the actuators in each of the ejectors are configured to actuate individually.
GPD003.11 Preferably, each of the ej ectors has a plurality of inlet channels extending from the reservoir to the chamber.
GPD003.12 Preferably, the spotting device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond-pads for connection to a microprocessor controller operatively controlling relative movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
GPD003.13 Preferably, the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
GPD003.14 Preferably, the spotting device also has a supporting substrate having a reservoir side and an ejector side opposite the reservoir side wherein the array of reservoirs is formed in the reservoir side and the array of ejectors is formed on the ejector side.
GPD003.15 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the surface with a density greater than 1 probe spot per square millimeter.
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GPD003.16 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the surface with a density greater than 8 probe spots per square millimeter.
GPD003.17 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the surface with a density greater than 60 probe spots per square millimeter.
GPD003.18 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the surface with a density between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GPD003.19 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the surface at a rate greater than 100 probe spots per second.
GPD003.20 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the surface at a rate greater than 1,400 probe spots per second. The mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them into the hybridization chambers of LOC devices that are being spotted. The data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides into the hybridization chambers of the LOC devices, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the
oligonucleotide specifications for storage into the memory of the LOC devices that are being spotted.
The oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy. The large numbers of oligonucleotide reservoirs and ejectors available on the oligonucleotide spotting device also provide for a one-step spotting of each LOC device.
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The data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GPD004.1 This aspect of the invention provides a biochemical deposition device for contactless deposition of biochemicals on a surface, the biochemical deposition device comprising:
an array of reservoirs for containing a plurality of biochemicals; and,
an array of ejectors, each of the ejectors being configured for fluid communication with a corresponding one of the reservoirs respectively; wherein,
the ejectors are configured to eject droplets containing the biochemical from the corresponding reservoir onto the surface.
GPD004.2 Preferably, the biochemicals in the array of reservoirs are oligonucleotide probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, and the surface is a lab-on-a- chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes.
GPD004.3 Preferably, the array of hybridization chambers is configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
GPD004.4 Preferably, the array of reservoirs has more than 1000 reservoirs. GPD004.5 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPD004.6 Preferably, the ejector has a chamber for containing liquid with suspended oligonucleotide probes supplied from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GPD004.7 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
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GPD004.8 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
GPD004.9 Preferably, the ejectors are configured to eject droplets having a volume less than 25 picoliters.
GPD004.10 Preferably, the ejectors are configured to eject droplets having a volume less than 6 picoliters.
GPD004.11 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
GPD004.12 Preferably, the actuators in each of the ejectors are configured to actuate individually.
GPD004.13 Preferably, each of the ejectors has a plurality of inlet channels extending from the reservoir to the chamber.
GPD004.14 Preferably, the biochemical deposition device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond- pads for connection to a microprocessor controller operatively controlling relative movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
GPD004.15 Preferably, the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
GPD004.16 Preferably, the biochemical deposition device also has a supporting substrate having a reservoir side and an ejector side opposite the reservoir side wherein the array of reservoirs is formed in the reservoir side and the array of ejectors is formed on the ejector side.
GPD004.17 Preferably, the array of ejectors is configured to eject droplets containing the biochemicals onto the surface with a density greater than 8 droplets per square millimeter.
GPD004.18 Preferably, the array of ejectors is configured to eject droplets containing the biochemicals onto the surface with a density greater than 60 droplets per square millimeter.
GPD004.19 Preferably, the array of ejectors is configured to eject droplets containing the biochemicals onto the surface with a density between 500 droplets per square millimeter and 1500 droplets per square millimeter.
GPD004.20 Preferably, the array of ejectors is configured to eject droplets containing the biochemicals onto the surface at a rate greater than 100 droplets per second.
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The mass-producible and inexpensive biochemical deposition device is used as a part of a cost-effective automated mass-manufacturing environment. Biochemicals are loaded in the device's biochemical reservoirs, and the device deposits them onto a surface by ejecting the biochemicals from its biochemical reservoir onto the surfaces being deposited upon. The data automation provided by the biochemical deposition device includes automated computer-controlled dispensing of the biochemicals onto the surface being spotted, receiving the specifications of the biochemicals stored in its reservoirs, storing the biochemicals specifications in its digital memory, and transmitting of the biochemicals specifications to segments of the automated manufacturing environment.
The biochemical deposition device provides for an automated, volumetrically and positionally precise, fast, and high-density biochemical deposition technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy. The data automation provided by the biochemical deposition device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GPD005.1 This aspect of the invention provides a microsystems technology (MST) device for contactless spotting of oligonucleotide probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the MST device comprising:
a monolithic substrate having a reservoir side and an ejector side opposite the reservoir side;
an array of reservoirs formed in the reservoir side; and,
an array of ejectors formed on the ejector side, each of the ejectors being configured for fluid communication with a corresponding one of the reservoirs respectively; wherein,
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the ejectors are configured to eject droplets containing the oligonucleotide probes from the corresponding reservoir onto the surface.
GPD005.2 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPD005.3 Preferably, the ejector has a chamber for containing liquid with suspended oligonucleotide probes supplied from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GPD005.4 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPD005.5 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
GPD005.6 Preferably, the ejectors are configured to eject droplets having a volume less than 25 picoliters.
GPD005.7 Preferably, the ejectors are configured to eject droplets having a volume less than 6 picoliters.
GPD005.8 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
GPD005.9 Preferably, the actuators in each of the ejectors are configured to actuate individually.
GPD005.10 Preferably, each of the ejectors has a plurality of inlet channels extending from the reservoir to the chamber.
GPD005.11 Preferably, the MST device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond-pads for connection to a microprocessor controller operatively controlling relative movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
GPD005.12 Preferably, the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
GPD005.13 Preferably, the surface is a lab-on-a-chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes, the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of
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reservoirs being configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
GPD005.14 Preferably, the array of reservoirs has more than 1000 reservoirs.
GPD005.15 Preferably, the CMOS circuitry is between the array of reservoirs and the array of ej ectors .
GPD005.16 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the surface with a density greater than 8 probe spots per square millimeter.
GPD005.17 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the surface with a density greater than 60 probe spots per square millimeter.
GPD005.18 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the surface with a density between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GPD005.19 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the surface at a rate greater than 100 probe spots per second.
GPD005.20 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the surface at a rate greater than 1,400 probe spots per second. The oligonucleotide spotting device is mass-produced inexpensively using microsystem technology (MST) and is used as a part of a cost-effective automated mass- manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted. The data automation provided by the oligonucleotide spotting device includes automated computer- controlled dispensing of the oligonucleotides onto the surface being spotted, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment. The oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids
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from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
The data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GPD006.1 This aspect of the invention provides an oligonucleotide spotting device for contactless spotting of oligonucleotide probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the oligonucleotide spotting device comprising:
an array of reservoirs for containing the oligonucleotide probes suspended in a liquid;
an array of ejectors overlaying the array of reservoirs, each of the ejectors being configured for fluid communication with a corresponding one of the reservoirs respectively; wherein,
the ejectors are configured to eject droplets containing the oligonucleotide probes from the corresponding reservoir onto the surface.
GPD006.2 Preferably, the oligonucleotide spotting device also has a supporting substrate having a reservoir side and an ejector side opposite the reservoir side wherein the array of reservoirs is formed in the reservoir side and the array of ejectors is formed on the ejector side.
GPD006.3 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPD006.4 Preferably, the ejector has a chamber for containing the liquid from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GPD006.5 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPD006.6 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
GPD006.7 Preferably, the ejectors are configured to eject droplets having a volume less than 25 picoliters.
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GPD006.8 Preferably, the ejectors are configured to eject droplets having a volume less than 6 picoliters.
GPD006.9 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
GPD006.10 Preferably, the actuators in each of the ejectors are configured to actuate individually.
GPD006.11 Preferably, each of the ejectors has a plurality of inlet channels extending from the reservoir to the chamber.
GPD006.12 Preferably, the oligonucleotide spotting device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond- pads for connection to a microprocessor controller operatively controlling relative movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
GPD006.13 Preferably, the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
GPD006.14 Preferably, the CMOS circuitry is between the array of reservoirs and the array of ejectors.
GPD006.15 Preferably, the surface is a lab-on-a-chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes, the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
GPD006.16 Preferably, the array of reservoirs has more than 1000 reservoirs. GPD006.17 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more than 8 probe spots per square millimeter.
GPD006.18 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more than 60 probe spots per square millimeter.
GPD006.19 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GPD006.20 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
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The mass-producible and inexpensive oligonucleotide spotting device with laminar structure is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted. The data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides onto the surfaces being spotted, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment.
The oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy. The data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GPD007.1 This aspect of the invention provides an oligonucleotide spotting device for contactless spotting of oligonucleotide probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the oligonucleotide spotting device comprising:
a supporting substrate;
an array of reservoirs on one side of the supporting substrate, the reservoirs configured for containing the oligonucleotide probes suspended in a liquid;
an array of ejectors on the other side of the supporting substrate; and,
a plurality of inlet channels for fluid communication between the reservoirs and the ejectors; wherein,
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the ejectors are configured to eject droplets containing the oligonucleotide probes from the corresponding reservoir onto the surface.
GPD007.2 Preferably, each of the ejectors is configured for fluid
communication with a corresponding one of the reservoirs respectively.
GPD007.3 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPD007.4 Preferably, the ejector has a chamber for containing the liquid from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GPD007.5 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPD007.6 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
GPD007.7 Preferably, the ejectors are configured to eject droplets having a volume less than 25 picoliters.
GPD007.8 Preferably, the ejectors are configured to eject droplets having a volume less than 6 picoliters.
GPD007.9 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
GPD007.10 Preferably, the actuators in each of the ejectors are configured to actuate individually.
GPD007.11 Preferably, each of the ejectors is in fluid communication with one of the reservoirs via more than one of the inlet channels.
GPD007.12 Preferably, the oligonucleotide spotting device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond- pads for connection to a microprocessor controller operatively controlling relative movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
GPD007.13 Preferably, the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
GPD007.14 Preferably, the CMOS circuitry is between the array of reservoirs and the array of ejectors.
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GPD007.15 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more than 8 probe spots per square millimeter.
GPD007.16 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more than 60 probe spots per square millimeter.
GPD007.17 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GPD007.18 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
GPD007.19 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 1,400 probe spots per second.
GPD007.20 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 20,000 probe spots per second.
The mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. It is fabricated with fluidics on both side of a silicon substrate, increasing the device integration level, reducing the device dimensions, and minimizing the device cost. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted. The data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides onto the surface being spotted, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment. The oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids
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from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
The data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GPD008.1 This aspect of the invention provides an oligonucleotide spotting device for contactless spotting of probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the oligonucleotide spotting device comprising:
a supporting substrate;
an array of ejectors, each having an actuator for ejecting droplets of liquid containing the oligonucleotide probes;
CMOS circuitry for providing each of the drop ejection actuators with a drive pulse for droplet ejection; and,
bond-pads for electrically connecting the CMOS circuitry and an external microprocessor controller for operative control of the array of ejectors.
GPD008.2 Preferably, the CMOS circuitry has a digital memory storing identity data for identifying the device to the external microprocessor controller.
GPD008.3 Preferably, the oligonucleotide spotting device also has an array of reservoirs for containing the oligonucleotide probes suspended in a liquid, wherein the supporting substrate has a reservoir side and an ejector side opposite the reservoir side, the array of reservoirs being formed in the reservoir side and, the array of ejectors formed on the ejector side, each of the ejectors being configured for fluid communication with a corresponding one of the probe reservoirs respectively.
GPD008.4 Preferably, the digital memory stores specification data for the oligonucleotide probes.
GPD008.5 Preferably, each of the ejectors has a plurality of the actuators and a corresponding plurality of nozzles associated with each of the droplet ejection actuators respectively, such that actuation of one of the actuators ejects a droplet through the nozzle associated with said actuator.
GPD008.6 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
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GPD008.7 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
GPD008.8 Preferably, the ejectors are configured to eject droplets having a volume less than 25 picoliters.
GPD008.9 Preferably, the ejectors are configured to eject droplets having a volume less than 6 picoliters.
GPD008.10 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
GPD008.11 Preferably, the actuators in one of the ejectors are configured to actuate individually.
GPD008.12 Preferably, each of the ejectors have a chamber for containing the liquid for ejection from the nozzle, and a plurality of inlet channels extending from the chamber to the reservoir corresponding to the ejector.
GPD008.13 Preferably, the actuators in each of the ejectors are configured to actuate individually.
GPD008.14 Preferably, the CMOS circuitry is between the array of reservoirs and the array of ejectors.
GPD008.15 Preferably, the surface is a lab-on-a-chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes, the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
GPD008.16 Preferably, the array of reservoirs has more than 1000 reservoirs. GPD008.17 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more than 8 probe spots per square millimeter.
GPD008.18 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more than 60 probe spots per square millimeter.
GPD008.19 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GPD008.20 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
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The mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted. The data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides onto the surface being spotted, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment. The spotting device performs these functions under external computer control.
The oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
The data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment. GPD009.1 This aspect of the invention provides an oligonucleotide spotting device for contactless spotting of probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the oligonucleotide spotting device comprising:
a supporting substrate;
an array of ejectors, each having an actuator for ejecting droplets of liquid containing the oligonucleotide probes; and,
CMOS circuitry for providing each of the drop ejection actuators with a drive pulse for droplet ejection; wherein,
the CMOS circuitry has a digital memory for storing data related to the device.
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GPD009.2 Preferably, the CMOS circuitry has bond-pads for electrically connecting and an external microprocessor controller for operative control of the array of ejectors.
GPD009.3 Preferably, the data includes identity data for identifying the device to the external microprocessor controller.
GPD009.4 Preferably, the oligonucleotide spotting device also has an array of reservoirs for containing the oligonucleotide probes suspended in a liquid, wherein the supporting substrate has a reservoir side and an ejector side opposite the reservoir side, the array of reservoirs being formed in the reservoir side and, the array of ejectors formed on the ejector side, each of the ejectors being configured for fluid communication with a corresponding one of the probe reservoirs respectively.
GPD009.5 Preferably, the digital memory stores specification data for the oligonucleotide probes.
GPD009.6 Preferably, each of the ejectors has a plurality of the actuators and a corresponding plurality of nozzles associated with each of the droplet ejection actuators respectively, such that actuation of one of the actuators ejects a droplet through the nozzle associated with said actuator.
GPD009.7 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPD009.8 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
GPD009.9 Preferably, the ejectors are configured to eject droplets having a volume less than 25 picoliters.
GPD009.10 Preferably, the ejectors are configured to eject droplets having a volume less than 6 picoliters.
GPD009.11 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
GPD009.12 Preferably, the actuators in one of the ejectors are configured to actuate individually.
GPD009.13 Preferably, each of the ejectors have a chamber for containing the liquid for ejection from the nozzle, and a plurality of inlet channels extending from the chamber to the reservoir corresponding to the ejector.
GPD009.14 Preferably, the actuators in each of the ejectors are configured to actuate individually.
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GPD009.15 Preferably, the CMOS circuitry is between the array of reservoirs and the array of ejectors.
GPD009.16 Preferably, the surface is a lab-on-a-chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes, the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
GPD009.17 Preferably, the array of reservoirs has more than 1000 reservoirs. GPD009.18 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more than 60 probe spots per square millimeter.
GPD009.19 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GPD009.20 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
The mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted. The data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides onto the surface being spotted, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment.
The oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids
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from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
The data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GPD010.1 This aspect of the invention provides a spotting device for contactless spotting of oligonucleotide probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the spotting device comprising:
an array of reservoirs for containing the oligonucleotide probes suspended in a liquid;
an array of ejectors in fluid communication with the reservoirs, each having an actuator for ejecting droplets of liquid containing the oligonucleotide probes; and,
CMOS circuitry for providing each of the drop ejection actuators with a drive pulse for droplet ejection; wherein,
the CMOS circuitry has a digital memory.
GPD010.2 Preferably, the CMOS circuitry has bond-pads for electrically connecting and an external microprocessor controller for operative control of the array of ejectors.
GPD010.3 Preferably, the digital memory stores identity data for identifying the device to the external microprocessor controller.
GPD010.4 Preferably, the oligonucleotide spotting device also has a supporting substrate with a reservoir side and an ejector side opposite the reservoir side, the array of reservoirs being formed in the reservoir side and, the array of ejectors formed on the ejector side, each of the ejectors being configured for fluid communication with a corresponding one of the probe reservoirs respectively.
GPD010.5 Preferably, the digital memory stores specification data for the oligonucleotide probes.
GPD010.6 Preferably, each of the ejectors has a plurality of the actuators and a corresponding plurality of nozzles associated with each of the droplet ejection actuators respectively, such that actuation of one of the actuators ejects a droplet through the nozzle associated with said actuator.
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GP DO 10.7 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPD010.8 Preferably, the ejectors are configured to eject droplets having a volume less than 2.0 picoliters.
GPD010.9 Preferably, the actuators in one of the ejectors are configured to actuate individually.
GPD010.10 Preferably, each of the ejectors have a chamber for containing the liquid for ejection from the nozzle, and a plurality of inlet channels extending from the chamber to the reservoir corresponding to the ejector.
GPD010.11 Preferably, the actuators in each of the ejectors are configured to actuate individually.
GPD010.12 Preferably, the CMOS circuitry is between the array of reservoirs and the array of ejectors.
GPD010.13 Preferably, the surface is a lab-on-a-chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes, the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
GPD010.14 Preferably, the array of reservoirs has more than 1000 reservoirs.
GPD010.15 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more than 8 probe spots per square millimeter.
GPD010.16 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more than 60 probe spots per square millimeter.
GPD010.17 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GPD010.18 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
GPD010.19 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 1,400 probe spots per second.
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GPDO 10.20 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 20,000 probe spots per second. The mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted. The data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides onto the surface being spotted, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment. The oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
The data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GPDO 11.1 This aspect of the invention provides a spotting device for contactless spotting of lab-on-a-chip (LOC) devices with oligonucleotide probes, the LOC devices being held in a fixed array on a printed circuit board (PCB) and each having an array of hybridization chambers for receiving the oligonucleotide probes, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the spotting device comprising:
a monolithic supporting substrate;
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an array of reservoirs on one side of the supporting substrate, the reservoirs containing sufficient amount of the oligonucleotide probes suspended in a liquid to spot all the LOC devices on the PCB; and,
an array of ejectors on the other side of the supporting substrate, each of the ejectors being configured for fluid communication with a corresponding one of the reservoirs respectively such that the ejectors eject droplets containing the oligonucleotide probes from the corresponding reservoir into one of the hybridization chambers.
GPD011.2 Preferably, the array of reservoirs has more than 1000 reservoirs.
GPD011.3 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPD011.4 Preferably, the ejector has a chamber for containing the liquid from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GP DO 11.5 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPD011.6 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
GPD011.7 Preferably, the ejectors are configured to eject droplets having a volume less than 25 picoliters.
GPD011.8 Preferably, the ejectors are configured to eject droplets having a volume less than 6 picoliters.
GPD011.9 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
GPD011.10 Preferably, the actuators in each of the ejectors are configured to actuate individually.
GPD011.11 Preferably, each of the ej ectors has a plurality of inlet channels extending from the reservoir to the chamber.
GPD011.12 Preferably, the spotting device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond-pads for connection to a microprocessor controller operatively controlling relative movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
GPD011.13 Preferably, the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
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GP DO 11.14 Preferably, the array of reservoirs is integrally formed into the one side of the monolithic supporting substrate.
GPDO 11.15 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more than 8 probe spots per square millimeter.
GPDO 11.16 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more than 60 probe spots per square millimeter.
GPDO 11.17 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GPD011.18 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
GPDO 11.19 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 1,400 probe spots per second.
GPDO 11.20 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 20,000 probe spots per second. The mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them into the hybridization chambers of the arrays of LOC devices mounted on PCB wafers. The data automation provided by the oligonucleotide spotting device includes automated computer- controlled spotting with oligonucleotide of the arrays of LOC devices mounted on PCB wafers, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to the LOC devices or other segments of the automated manufacturing environment.
The oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The device also functions
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as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy. The data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
Spotting with oligonucleotide of the arrays of LOC devices mounted on PCB wafers speeds up and reduces the cost of the loading process, and spotting the LOC devices after mounting them on the PCB wafers and soldering them, improves the chemical and physical integrity of the oligonucleotide.
GP DO 12.1 This aspect of the invention provides an oligonucleotide spotting device for contactless spotting of a silicon wafer on which an array of lab-on-a-chip (LOC) devices are fabricated, the LOC devices being configured to use the oligonucleotide probes to detect target nucleic acid sequences in a biological sample and each having an array of hybridization chambers for receiving the oligonucleotide probes, the oligonucleotide spotting device comprising:
an array of reservoirs on one side of the supporting substrate, the reservoirs containing sufficient amount of the oligonucleotide probes suspended in a liquid to spot all the LOC devices on the wafer; and,
an array of ejectors overlaying the array of reservoirs for fixed movement therewith, such that the ejectors eject droplets containing the oligonucleotide probes from the corresponding reservoir into one of the hybridization chambers.
GPD012.2 Preferably, the oligonucleotide spotting device also has a supporting substrate having a reservoir side and an ejector side opposite the reservoir side wherein the array of reservoirs is formed in the reservoir side and the array of ejectors is formed on the ejector side.
GPD012.3 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPD012.4 Preferably, the ejector has a chamber for containing the liquid from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding
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to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GP DO 12.5 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPD012.6 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
GPD012.7 Preferably, the ejectors are configured to eject droplets having a volume less than 25 picoliters.
GPD012.8 Preferably, the ejectors are configured to eject droplets having a volume less than 6 picoliters.
GPD012.9 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
GPD012.10 Preferably, the actuators in each of the ejectors are configured to actuate individually.
GPD012.11 Preferably, each of the ejectors has a plurality of inlet channels extending from the reservoir to the chamber.
GPD012.12 Preferably, the oligonucleotide spotting device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond- pads for connection to a microprocessor controller operatively controlling relative movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
GPD012.13 Preferably, the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
GPD012.14 Preferably, the CMOS circuitry is between the array of reservoirs and the array of ej ectors .
GPD012.15 Preferably, the LOC device has an array of hybridization chambers for receiving the oligonucleotide probes, the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a
predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
GPD012.16 Preferably, the array of reservoirs has more than 1000 reservoirs.
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GPD012.17 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GPD012.18 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
GPD012.19 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 1,400 probe spots per second.
GPD012.20 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 20,000 probe spots per second.
The mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them into the hybridization chambers of the arrays of LOC devices on partial-depth sawn wafers. The data automation provided by the oligonucleotide spotting device includes automated computer-controlled spotting with oligonucleotide of the arrays of LOC devices on partial- depth sawn wafers, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to the LOC devices or other segments of the automated manufacturing environment. The oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
GCA003-PCT
The data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment. Spotting with oligonucleotide of the arrays of LOC devices on partial-depth sawn wafers speeds up and reduces the cost of the loading process.
GPDO 13.1 This aspect of the invention provides an oligonucleotide spotting device for contactless spotting of probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the oligonucleotide spotting device comprising:
a monolithic supporting substrate;
an array of ejectors formed on one side of the supporting substrate such that the ejectors eject droplets containing the oligonucleotide probes onto the surface; wherein during use,
each of the ejectors in the array is configured to eject droplets having a volume less than 100 picoliters.
GPD013.2 Preferably, each of the ejectors in the array is configured to eject droplets having a volume less than 25 picoliters.
GPDO 13.3 Preferably, each of the ejectors in the array is configured to eject droplets having a volume less than 6 picoliters.
GPD013.4 Preferably, each of the ejectors in the array is configured to eject droplets having a volume between 0.1 picoliter and 1.6 picoliters.
GPDO 13.5 Preferably, the monolithic supporting substrate has a reservoir side and an ejection side opposite the reservoir side, the array of ejectors being formed on the ejection side and an array of reservoirs formed in the reservoir side, each of the ejectors being configured for fluid communication with a corresponding one of the probe reservoirs respectively.
GPDO 13.6 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPDO 13.7 Preferably, the ejector has a chamber for containing the liquid supplied from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
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GPD013.8 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPD013.9 Preferably, the actuators in one of the ejectors are configured to actuate individually.
GPDO 13.10 Preferably, each of the ej ectors has a plurality of inlet channels extending from the reservoir to the chamber.
GPD013.11 Preferably, the oligonucleotide spotting device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond- pads for connection to a control microprocessor operatively controlling relative movement between the nozzles and the surface to be spotted with the probes.
GPD013.12 Preferably, the CMOS circuitry has memory for storing specification data relating to the probes in the reservoirs.
GPDO 13.13 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more than 8 probe spots per square millimeter.
GPDO 13.14 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more than 60 probe spots per square millimeter.
GPDO 13.15 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GPDO 13.16 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
GPDO 13.17 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 1,400 probe spots per second.
GPDO 13.18 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 20,000 probe spots per second.
GPDO 13.19 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate between 300,000 probe spots per second and 1,000,000 probe spots per second.
GPDO 13.20 Preferably, the array of reservoirs are integrally formed in the reservoir side of the monolithic substrate.
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The mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted. The data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides onto the surface being spotted, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment.
The oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy. In particular, the capability of the device to spot the requisite low-volume probes provides for low probe cost, in turn, permitting the inexpensive assay system.
The data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment. GPD014.1 This aspect of the invention provides an oligonucleotide spotting device for contactless spotting of probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the oligonucleotide spotting device comprising:
an array of ejectors, each having an actuator for ejecting droplets of liquid containing the probes;
CMOS circuitry for providing each of the actuators with a drive pulse for droplet ejection; wherein during use,
the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
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GPD014.2 Preferably, the array of ejectors spot the probes onto the surface at a rate greater than 1,400 probe spots per second.
GPD014.3 Preferably, the array of ejectors spot the probes onto the surface at a rate greater than 20,000 probe spots per second.
GPD014.4 Preferably, the array of ejectors spot the probes onto the surface at a rate between 300,000 probe spots per second and 1,000,000 probe spots per second.
GPD014.5 Preferably, the CMOS circuitry has bond-pads for connection to an external control microprocessor for operative control of the array of ejectors.
GPD014.6 Preferably, the CMOS circuitry has a digital memory storing identity data for identifying the device to the external microprocessor controller.
GPD014.7 Preferably, the digital memory stores probe type data and probe location data, the probe type data identifying the probe types in the device and the probe location data identifying the reservoir location for each of the probe types.
GPD014.8 Preferably, the oligonucleotide spotting device also has a supporting substrate, the supporting substrate has a reservoir side and an ejector side opposite the reservoir side, the array of ejectors being formed on the ejector side and an array of reservoirs being formed in the reservoir side, each of the ejectors being configured for fluid communication with a corresponding one of the probe reservoirs respectively for ejecting droplets containing the probes from the corresponding reservoir onto the surface.
GPD014.9 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPD014.10 Preferably, the ejector has a chamber for containing liquid supplied from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GP DO 14.11 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPD014.12 Preferably, the actuators in one of the ejectors are configured to actuate individually.
GPD014.13 Preferably, each of the ejectors has a plurality of inlet channels extending from the reservoir to the chamber.
GPD014.14 Preferably,
the surface is a lab-on-a-chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes, the array of hybridization chambers being configured
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to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
GPD014.15 Preferably, the array of reservoirs has more than 1000 reservoirs.
GPD014.16 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
GPD014.17 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 1,400 probe spots per second.
GPD014.18 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 20,000 probe spots per second.
GPD014.19 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate between 300,000 probe spots per second and 1,000,000 probe spots per second.
GPD014.20 Preferably, the array of reservoirs are integrally formed in the reservoir side of the monolithic substrate.
The mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted. The data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides onto the surface being spotted, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment.
The oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The high spotting rate of
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the device, in turn, provides a high spotting throughput and reduces the overall cost of the product assay system. The device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
The data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment. GPD015.1 This aspect of the invention provides a biochemical deposition device for contactless deposition of biochemicals on a surface, the biochemical deposition device comprising:
a supporting substrate;
an array of reservoirs on one side of the substrate, the reservoirs being configured for containing a plurality of biochemicals; and,
an array of ejectors on the other side of the supporting substrate, each of the ejectors being configured for fluid communication with the reservoirs; wherein during use, the array of ejectors eject droplets containing the biochemicals onto the surface at a rate greater than 100 droplets per second.
GPD015.2 Preferably, the array of ejectors eject droplets containing the biochemicals onto the surface at a rate greater than 1,400 droplets per second.
GPD015.3 Preferably, the array of ejectors eject droplets containing the biochemicals onto the surface at a rate greater than 20,000 droplets per second.
GPD015.4 Preferably, the array of ejectors eject droplets containing the biochemicals onto the surface at a rate between 300,000 droplets per second and 1,000,000 droplets per second.
GPD015.5 Preferably, the biochemicals in the array of reservoirs are oligonucleotide probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, and the surface is a lab-on-a- chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes.
GPD015.6 Preferably, the array of hybridization chambers is configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay
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of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
GPD015.7 Preferably, the array of reservoirs has more than 1000 reservoirs.
GPD015.8 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPD015.9 Preferably, the ejector has a chamber for containing liquid with suspended oligonucleotide probes supplied from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GP DO 15.10 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPD015.11 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
GPD015.12 Preferably, the ejectors are configured to eject droplets having a volume less than 25 picoliters.
GPD015.13 Preferably, the ejectors are configured to eject droplets having a volume less than 6 picoliters.
GPD015.14 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
GPD015.15 Preferably, the actuators in each of the ejectors are configured to actuate individually.
GPD015.16 Preferably, each of the ej ectors has a plurality of inlet channels extending from the reservoir to the chamber.
GP DO 15.17 Preferably, the biochemical deposition device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond- pads for connection to a microprocessor controller operatively controlling relative movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
GPD015.18 Preferably, the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
GPD015.19 Preferably, the array of ejectors is configured to eject droplets containing the biochemicals onto the surface with a density greater than 8 droplets per square millimeter.
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GPD015.20 Preferably, the array of ejectors is configured to eject droplets containing the biochemicals onto the surface with a density greater than 60 droplets per square millimeter. The mass-producible and inexpensive biochemical deposition device is used as a part of a cost-effective automated mass-manufacturing environment. Biochemicals are loaded in the device's biochemical reservoirs, and the device deposits them onto a surface by ejecting the biochemicals from its biochemical reservoir onto the surfaces being deposited upon. The data automation provided by the biochemical deposition device includes automated computer-controlled dispensing of the biochemicals onto the surface being spotted, receiving the specifications of the biochemicals stored in its reservoirs, storing the biochemicals specifications in its digital memory, and transmitting of the biochemicals specifications to segments of the automated manufacturing environment. The biochemical deposition device provides for an automated, volumetrically and positionally precise, fast, and high-density biochemical deposition technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The high deposition rate of the device, in turn, provides a high deposition throughput and reduces the overall product costs. The device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy.
The data automation provided by the biochemical deposition device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GPD016.1 This aspect of the invention provides an oligonucleotide spotting device for contactless spotting of probes onto a surface, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, the oligonucleotide spotting device comprising:
an array of ejectors, each having an actuator for ejecting droplets of liquid containing the probes;
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CMOS circuitry for providing each of the actuators with a drive pulse for droplet ejection; wherein,
the array of ejectors is configured to spot the probes onto the surface at a density more than 1 probe spot per square millimeter.
GPDO 16.2 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more than 8 probe spots per square millimeter.
GPDO 16.3 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more than 60 probe spots per square millimeter.
GPDO 16.4 Preferably, the array of ejectors is configured to spot the probes onto the surface at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GPD016.5 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 100 probe spots per second.
GPD016.6 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 1,400 probe spots per second.
GPD016.7 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate greater than 20,000 probe spots per second.
GPDO 16.8 Preferably, the CMOS circuitry is configured to generate drive pulses such the array of ejectors spot the probes onto the surface at a rate between 300,000 probe spots per second and 1,000,000 probe spots per second.
GPD016.9 Preferably, the CMOS circuitry has bond-pads for connection to an external control microprocessor for operative control of the array of ejectors.
GPD016.10 Preferably, the CMOS circuitry has a digital memory storing identity data for identifying the device to the external microprocessor controller.
GPDO 16.11 Preferably, the digital memory stores probe type data and probe location data, the probe type data identifying the probe types in the device and the probe location data identifying the reservoir location for each of the probe types.
GPDO 16.12 Preferably, the oligonucleotide spotting device also has a supporting substrate, the supporting substrate has a reservoir side and an ejector side opposite the reservoir side, the array of ejectors being formed on the ejector side and an array of reservoirs being formed in the reservoir side, each of the ejectors being configured for fluid
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communication with a corresponding one of the probe reservoirs respectively for ejecting droplets containing the probes from the corresponding reservoir onto the surface.
GPD016.13 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPD016.14 Preferably, the ejector has a chamber for containing liquid supplied from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GPD016.15 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPD016.16 Preferably, the actuators in one of the ejectors are configured to actuate individually.
GPDO 16.17 Preferably, each of the ej ectors has a plurality of inlet channels extending from the reservoir to the chamber.
GPDO 16.18 Preferably,
the surface is a lab-on-a-chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes, the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
GPD016.19 Preferably, the array of reservoirs has more than 1000 reservoirs. GPDO 16.20 Preferably, the array of reservoirs are integrally formed in the reservoir side of the monolithic substrate.
The mass-producible and inexpensive oligonucleotide spotting device is used as a part of a cost-effective automated mass-manufacturing environment. Oligonucleotides are loaded in the device's oligonucleotides reservoirs, and the device ejects them onto the surfaces that are being spotted. The data automation provided by the oligonucleotide spotting device includes automated computer-controlled dispensing of the oligonucleotides onto the surface being spotted, receiving the specifications of the oligonucleotides stored in its reservoirs, storing the oligonucleotide specifications in its digital memory, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment.
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The oligonucleotide spotting device provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy. In particular, the capability of the device to spot at the requisite high-density provides for low final product dimensions, in turn, permitting the inexpensive assay system.
The data automation provided by the oligonucleotide spotting device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GPD017.1 This aspect of the invention provides a biochemical deposition device for contactless deposition of biochemicals on a surface, the biochemical deposition device comprising:
a supporting substrate;
an array of reservoirs on one side of the substrate, the reservoirs being configured for containing a plurality of biochemicals; and,
an array of ejectors on the other side of the supporting substrate, the ejectors being in fluid communication with the reservoirs, and configured to eject droplets containing the biochemicals onto the surface with a density greater than 1 droplet per square millimeter.
GPD017.2 Preferably, the array of ejectors is configured to eject droplets containing the biochemicals onto the surface with a density greater than 8 droplets per square millimeter.
GPD017.3 Preferably, the array of ejectors is configured to eject droplets containing the biochemicals onto the surface with a density greater than 60 droplets per square millimeter.
GPD017.4 Preferably, the array of ejectors is configured to eject droplets containing the biochemicals onto the surface with a density between 500 droplets per square millimeter and 1500 droplets per square millimeter.
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GPD017.5 Preferably, the array of ejectors is configured to eject droplets containing the biochemicals onto the surface at a rate greater than 100 droplets per second.
GPD017.6 Preferably, the array of ejectors is configured to eject droplets containing the biochemicals onto the surface at a rate greater than 1,400 droplets per second.
GPD017.7 Preferably, the array of ejectors is configured to eject droplets containing the biochemicals onto the surface at a rate greater than 20,000 droplets per second.
GPD017.8 Preferably, the array of ejectors is configured to eject droplets containing the biochemicals onto the surface at a rate between 300,000 droplets per second and 1,000,000 droplets per second.
GPD017.9 Preferably, the biochemicals in the array of reservoirs are oligonucleotide probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample, and the surface is a lab-on-a- chip (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probes.
GPD017.10 Preferably, the array of hybridization chambers is configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
GPD017.11 Preferably, the array of reservoirs has more than 1000 reservoirs.
GPD017.12 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPD017.13 Preferably, the ejector has a chamber for containing liquid with suspended oligonucleotide probes supplied from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GPD017.14 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPD017.15 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
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GPD017.16 Preferably, the actuators in each of the ejectors are configured to actuate individually.
GPD017.17 Preferably, each of the ejectors has a plurality of inlet channels extending from the reservoir to the chamber.
GPD017.18 Preferably, the biochemical deposition device also has CMOS circuitry for providing the actuators with drive pulses, the CMOS circuitry having bond- pads for connection to a microprocessor controller operatively controlling relative movement between the nozzles and the surface to be spotted with the oligonucleotide probes.
GPD017.19 Preferably, the CMOS circuitry has memory for storing specification data related to the oligonucleotide probes in the reservoir.
GPD017.20 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters. The mass-producible and inexpensive biochemical deposition device is used as a part of a cost-effective automated mass-manufacturing environment. Biochemicals are loaded in the device's biochemical reservoirs, and the device deposits them onto a surface by ejecting the biochemicals from its biochemical reservoir onto the surfaces being deposited upon. The data automation provided by the biochemical deposition device includes automated computer-controlled dispensing of the biochemicals onto the surface being spotted, receiving the specifications of the biochemicals stored in its reservoirs, storing the biochemicals specifications in its digital memory, and transmitting of the biochemicals specifications to segments of the automated manufacturing environment. The biochemical deposition device provides for an automated, volumetrically and positionally precise, fast, and high-density biochemical deposition technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. The device also functions as the intermediate fluidic manipulation mechanism that is required for transferring fluids from a macroscopic level of volume and positioning accuracy to a microscopic level of volume and positioning accuracy. In particular, the capability of the device to deposit biochemicals at the requisite high-density provides for low final product dimensions, in turn, permitting the inexpensive product.
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The data automation provided by the biochemical deposition device provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment. GAL001.1 This aspect of the invention provides a robotic system for spotting oligonucleotides comprising:
an oligonucleotide spotting device for contactless spotting of oligonucleotides onto a surface, the oligonucleotide spotting device having an array of ejectors, each having a drop ejection actuator for ejecting droplets of liquid containing the oligonucleotides onto a surface, at least one reservoir in fluid communication with one or more of the ejectors and CMOS drive circuitry for providing each of the drop ejection actuators with a drive pulse for droplet ejection; and,
an apparatus for loading oligonucleotides into the oligonucleotide spotting device, the apparatus having a stage for detachably mounting a plurality of the oligonucleotide spotting devices, and a plurality of oligonucleotide containers mounted for movement relative to the stage, each of the oligonucleotide containers having a droplet dispenser for ejecting droplets of fluid containing oligonucleotides into the reservoirs of the oligonucleotide spotting devices.
GAL001.2 Preferably, the oligonucleotide spotting device has bond-pads for electrically connecting the CMOS drive circuitry and the apparatus such that the apparatus downloads oligonucleotide data to memory within the CMOS drive circuitry.
GAL001.3 Preferably, the apparatus has a camera for optically aligning the stage relative to the droplet dispensers.
GAL001.4 Preferably, the oligonucleotide spotting device has a supporting substrate for supporting the CMOS circuitry, the supporting substrate having a reservoir side in which the at least one reservoir is formed and an ejector side opposite the reservoir side, in which the array of ejectors are formed.
GAL001.5 Preferably, the oligonucleotide spotting device has an array of the reservoirs in the reservoir side, each of the reservoirs being in fluid communication with two or more of the ejectors.
GAL001.6 Preferably, the oligonucleotides are configured to be probes for hybridization with target nucleic acid sequences in a biological sample, and the liquid is a probe solution such that each of the ejectors have a respective nozzle through which the drop ejection actuator ejects a droplet of the probe solution.
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GAL001.7 Preferably, the oligonucleotide spotting device has at least one common chamber for containing the fluid to be ejected by a plurality of the nozzles.
GAL001.8 Preferably, each of the common chambers has a plurality of chamber inlets in fluid communication with the reservoir.
GAL001.9 Preferably, the oligonucleotide spotting device has an array of the reservoirs and an array of the common chambers respectively in fluid communication with one of the reservoirs.
GAL001.10 Preferably, the drop ejection actuators each have a heater for generating a vapor bubble in the fluid to eject a droplet through the nozzle corresponding to that drop ejection actuator.
GAL001.1 1 Preferably, the ejectors are each configured to eject a droplet of the probe solution having a volume less than 100 picoliters.
GAL001.12 Preferably, the ejectors are each configured to eject a droplet of the probe solution having a volume less than 25 picoliters.
GAL001.13 Preferably, the ejectors are each configured to eject a droplet of the probe solution having a volume less than 6 picoliters.
GAL001.14 Preferably, the ejectors are each configured to eject a droplet of the probe solution having a volume between 0.1 picoliters and 1.6 picoliters.
GAL001.15 Preferably, the containers are vials for containing an aliquot of the probe solution, each of the vials having a quality assurance chip with memory for storing data identifying the oligonucleotides.
GAL001.16 Preferably, the vials each have a droplet dispenser and electrical contacts for receiving an actuation signal to activate the droplet dispenser.
GAL001.17 Preferably, the droplet dispenser has a piezoelectric actuator.
GAL001.18 Preferably, the apparatus is configured for transferring data from the quality assurance chip of the vials to the CMOS circuitry of the oligonucleotide spotting device.
GAL001.19 Preferably, the vials are suspended on a rack adj acent the oligonucleotide spotting devices on one side of the stage, the rack being configured to establish an individual electrical connection with each of the vials.
The apparatus for loading of oligonucleotide spotting devices is used, as part of a cost-effective automated mass-manufacturing environment, to dispense oligonucleotides contained in oligonucleotide microvials into the oligonucleotide reservoirs of
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oligonucleotide spotting devices. The data automation provided by the apparatus includes automated computer-controlled dispensing of the oligonucleotides into the oligonucleotide reservoirs of the oligonucleotide spotting devices, checking the oligonucleotide data stored in the memory of the microvials against the list of specifications for the oligonucleotides that have to be loaded in the oligonucleotide spotting devices, and storage of the oligonucleotide data into the memory of the oligonucleotide spotting devices.
The apparatus for loading of oligonucleotide spotting devices provides for an automated and volumetrically and positionally precise oligonucleotide dispensing technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
The data automation provided by the apparatus for loading of oligonucleotide spotting devices provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GPA001.1 This aspect of the invention provides an oligonucleotide spotting robot for spotting oligonucleotide probes into a microfluidic device having a digital memory for data related to the oligonucleotide probes loaded into the microfluidic device, the oligonucleotide dispensing robot comprising:
an array of reservoirs for containing the oligonucleotide probes suspended in a liquid;
an array of ejectors, each of the ejectors being configured for fluid communication with a corresponding one of the reservoirs respectively;
a mounting surface for detachably mounting the microfluidic device for movement relative to the ejectors; and,
a control processor for operative control of the ejectors and the mounting surface; wherein,
the control processor is configured to activate the ejectors, move the ejectors selected for activation into registration with the microfluidic device and download the data to the digital memory.
GPA001.2 Preferably, the oligonucleotide spotting robot also has a camera for optical feedback of the registration between the ejectors selected by the control processor and the microfluidic device.
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GPA001.3 Preferably, the array of reservoirs has more than 1000 reservoirs.
GPA001.4 Preferably, the oligonucleotide spotting robot also has CMOS circuitry between the array of reservoirs and the array of ejectors, the CMOS circuitry being configured to drive the array of ejectors in accordance with control signals from the control microprocessor, wherein the CMOS circuitry stores the data relating to the oligonucleotide probes.
GPA001.5 Preferably, the mounting surface is a stage configured for movement along two orthogonal axes, and the array of ejectors is mounted closely adjacent to, and facing, the stage.
GPA001.6 Preferably, the micro fluidic device is a lab-on-a-chip (LOC) device, the LOC device having an array of hybridization chambers for receiving the
oligonucleotide probes, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample and the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs being configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
GPA001.7 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPA001.8 Preferably, the ejector has a chamber for containing the liquid from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GPA001.9 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPA001.10 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
GPA001.11 Preferably, the ejectors are configured to eject droplets having a volume less than 25 picoliters.
GPA001.12 Preferably, the ejectors are configured to eject droplets having a volume less than 6 picoliters.
GPA001.13 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
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GPA001.14 Preferably, the actuators in each of the ejectors are configured to actuate individually.
GP A001.15 Preferably, each of the ej ectors has a plurality of inlet channels extending from the reservoir to the chamber.
GPA001.16 Preferably, the array of ejectors is configured to spot the oligonucleotide probes onto the LOC device with a density greater than 1 probe spot per square millimeter.
GPA001.17 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the LOC device with a density greater than 8 probe spots per square millimeter.
GPA001.18 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the LOC device with a density greater than 60 probe spots per square millimeter.
GPA001.19 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the LOC device with a density between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GPA001.20 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the LOC device at a rate greater than 100 probe spots per second. The oligonucleotide spotting robot is used as part of a cost-effective automated mass-manufacturing environment. Loaded oligonucleotide spotting devices are picked up by the robot and the robot positions the oligonucleotide spotting devices and commands them to eject the oligonucleotides onto the surfaces that are being spotted. The data automation provided by the oligonucleotide spotting robot includes automated computer- controlled dispensing of the oligonucleotides onto the surface being spotted, reading the specifications of the oligonucleotides stored in the reservoirs of the oligonucleotide spotting devices, checking the specifications of the oligonucleotides stored in the reservoirs of the oligonucleotide spotting devices against the relevant databases, storing the oligonucleotide specifications in the memory of the devices that are being spotted, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment.
The oligonucleotide spotting robot provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying
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the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
The data automation provided by the oligonucleotide spotting robot provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GPA003.1 This aspect of the invention provides an oligonucleotide spotting robot for spotting oligonucleotide probes into an array of lab-on-a-chip (LOC) devices, each having a digital memory for data related to the oligonucleotide probes loaded into that LOC device, the oligonucleotide dispensing robot comprising:
an array of reservoirs for containing the oligonucleotide probes suspended in a liquid;
an array of ejectors, each of the ejectors being configured for fluid communication with a corresponding one of the reservoirs respectively;
a mounting surface for detachably mounting the array of LOC devices for movement relative to the ejectors; and,
a control processor for operative control of the ejectors and the mounting surface; wherein,
the control processor is configured to activate the ejectors, move the ejectors selected for activation into registration with one or more of the LOC devices and download the data specifically relevant to each of the LOC devices into the digital memory of that LOC device.
GPA003.2 Preferably, the oligonucleotide spotting robot also has a camera for optical feedback of the registration between the ejectors selected by the control processor and the LOC devices.
GPA003.3 Preferably, the array of reservoirs has more than 1000 reservoirs.
GPA003.4 Preferably, the oligonucleotide spotting robot also has CMOS circuitry between the array of reservoirs and the array of ejectors, the CMOS circuitry being configured to drive the array of ejectors in accordance with control signals from the control microprocessor, wherein the CMOS circuitry stores the data relating to the oligonucleotide probes.
GPA003.5 Preferably, the array of LOC devices are mounted to a printed circuit board (PCB) which is in turn detachably mounted to the mounting surface, the mounting
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surface being a stage configured for movement along two orthogonal axes, and the array of ejectors is mounted closely adjacent to, and facing, the stage.
GPA003.6 Preferably, each of the LOC devices has an array of hybridization chambers for receiving the oligonucleotide probes, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample and the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs being configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC devices.
GPA003.7 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPA003.8 Preferably, the ejector has a chamber for containing the liquid from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GPA003.9 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPA003.10 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
GPA003.11 Preferably, the ejectors are configured to eject droplets having a volume less than 25 picoliters.
GPA003.12 Preferably, the ejectors are configured to eject droplets having a volume less than 6 picoliters.
GPA003.13 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
GPA003.14 Preferably, the actuators in each of the ejectors are configured to actuate individually.
GP A003.15 Preferably, each of the ej ectors has a plurality of inlet channels extending from the reservoir to the chamber.
GPA003.16 Preferably, the array of ejectors is configured to spot the oligonucleotide probes onto the LOC device with a density greater than 1 probe spot per square millimeter.
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GPA003.17 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the LOC device with a density greater than 8 probe spots per square millimeter.
GPA003.18 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the LOC device with a density greater than 60 probe spots per square millimeter.
GPA003.19 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the LOC device with a density between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GPA003.20 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the LOC device at a rate greater than 100 probe spots per second.
The oligonucleotide spotting robot is used as part of a cost-effective automated mass-manufacturing environment. Loaded oligonucleotide spotting devices are picked up by the robot and the robot positions the oligonucleotide spotting devices and commands them to eject the oligonucleotides into the hybridization chambers of the arrays of LOC devices mounted on PCB wafers. The data automation provided by the oligonucleotide spotting robot includes automated computer-controlled dispensing of the oligonucleotides into the hybridization chambers of the arrays of LOC devices mounted on PCB wafers, reading the specifications of the oligonucleotides stored in the reservoirs of the oligonucleotide spotting devices, checking the specifications of the oligonucleotides stored in the reservoirs of the oligonucleotide spotting devices against the relevant databases, storing the oligonucleotide specifications in the memory of the LOC devices that are being spotted, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment.
The oligonucleotide spotting robot provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
The data automation provided by the oligonucleotide spotting robot provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
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Spotting with oligonucleotide of the arrays of LOC devices mounted on PCB wafers speeds up and reduces the cost of the loading process, and spotting the LOC devices after mounting them on the PCB wafers and soldering them, improves the chemical and physical integrity of the oligonucleotide.
GPA004.1 This aspect of the invention provides an oligonucleotide spotting robot for spotting oligonucleotide probes into a silicon wafer on which an array of lab-on- a-chip (LOC) devices are fabricated, each LOC device having a digital memory for data related to the reagents loaded into the LOC device, the oligonucleotide dispensing robot comprising:
an array of reservoirs for containing the oligonucleotide probes suspended in a liquid;
an array of ejectors, each of the ejectors being configured for fluid communication with a corresponding one of the reservoirs respectively;
a mounting surface for detachably mounting the array of LOC devices for movement relative to the ejectors; and,
a control processor for operative control of the ejectors and the mounting surface; wherein,
the control processor is configured to activate the ejectors, move the ejectors selected for activation into registration with one or more of the LOC devices and download the data specifically relevant to each of the LOC devices into the digital memory of that LOC device.
GPA004.2 Preferably, the oligonucleotide spotting robot also has a camera for optical feedback of the registration between the ejectors selected by the control processor and the LOC devices.
GPA004.3 Preferably, the array of reservoirs has more than 1000 reservoirs.
GPA004.4 Preferably, the oligonucleotide spotting robot also has CMOS circuitry between the array of reservoirs and the array of ejectors, the CMOS circuitry being configured to drive the array of ejectors in accordance with control signals from the control microprocessor, wherein the CMOS circuitry stores the data relating to the oligonucleotide probes.
GPA004.5 Preferably, the silicon wafer is partially sawn in preparation for dicing into individually separate LOC devices, and the silicon wafer being detachably
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mounted to the mounting surface, the mounting surface being a stage configured for movement along two orthogonal axes, and the array of ejectors is mounted closely adjacent to, and facing, the stage.
GPA004.6 Preferably, each of the LOC devices has an array of hybridization chambers for receiving the oligonucleotide probes, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample and the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs being configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC devices.
GPA004.7 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPA004.8 Preferably, the ejector has a chamber for containing the liquid from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GPA004.9 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPA004.10 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
GPA004.11 Preferably, the ejectors are configured to eject droplets having a volume less than 25 picoliters.
GPA004.12 Preferably, the ejectors are configured to eject droplets having a volume less than 6 picoliters.
GPA004.13 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
GPA004.14 Preferably, the actuators in each of the ejectors are configured to actuate individually.
GPA004.15 Preferably, each of the ejectors has a plurality of inlet channels extending from the reservoir to the chamber.
GPA004.16 Preferably, the array of ejectors is configured to spot the oligonucleotide probes onto the LOC device with a density greater than 1 probe spot per square millimeter.
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GPA004.17 Preferably, the array of ejectors is configured to spot the
oligonucleotides onto the LOC device with a density greater than 8 probe spots per square millimeter.
GPA004.18 Preferably, the array of ejectors is configured to spot the
oligonucleotides onto the LOC device with a density greater than 60 probe spots per square millimeter.
GPA004.19 Preferably, the array of ejectors is configured to spot the
oligonucleotides onto the LOC device with a density between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GPA004.20 Preferably, the array of ejectors is configured to spot the
oligonucleotides onto the LOC device at a rate greater than 100 probe spots per second.
The oligonucleotide spotting robot is used as part of a cost-effective automated mass-manufacturing environment. Loaded oligonucleotide spotting devices are picked up by the robot and the robot positions the oligonucleotide spotting devices and commands them to eject the oligonucleotides into the hybridization chambers of the arrays of LOC devices on partial-depth sawn wafers. The data automation provided by the oligonucleotide spotting robot includes automated computer-controlled dispensing of the oligonucleotides into the hybridization chambers of the arrays of LOC devices on partial-depth sawn wafers, reading the specifications of the oligonucleotides stored in the reservoirs of the oligonucleotide spotting devices, checking the specifications of the oligonucleotides stored in the reservoirs of the oligonucleotide spotting devices against the relevant databases, storing the oligonucleotide specifications in the memory of the LOC devices that are being spotted, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment.
The oligonucleotide spotting robot provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
The data automation provided by the oligonucleotide spotting robot provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
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Spotting with oligonucleotide of the arrays of LOC devices on partial-depth sawn wafers speeds up and reduces the cost of the loading process. GPA005.1 This aspect of the invention provides an oligonucleotide spotting robot for spotting oligonucleotide probes onto a substrate, the oligonucleotide dispensing robot comprising:
an array of reservoirs for containing the oligonucleotide probes suspended in a liquid;
an array of ejectors, each of the ejectors being configured for fluid communication with a corresponding one of the reservoirs respectively;
a mounting surface for detachably mounting the substrate for movement relative to the ejectors; and,
a control processor for operative control of the ejectors and the mounting surface; wherein,
the array of ejectors is configured to spot the probes onto the surface at a density more than 1 probe spot per square millimeter.
GPA005.2 Preferably, the array of ejectors is configured to spot the probes onto the substrate at a density more than 8 probe spots per square millimeter.
GPA005.3 Preferably, the array of ejectors is configured to spot the probes onto the substrate at a density more than 60 probe spots per square millimeter.
GPA005.4 Preferably, the array of ejectors is configured to spot the probes onto the substrate at a density more between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GPA005.5 Preferably, the substrate is a lab-on-a-chip (LOC) device with a digital memory for data related to the oligonucleotide probes loaded into the LOC device, and the control microprocessor is configured to activate the ejectors, move the ejectors selected for activation into registration with the LOC device and download the data to the digital memory.
GPA005.6 Preferably, the oligonucleotide spotting robot also has a camera for optical feedback of the registration between the ejectors selected by the control processor and the LOC device.
GPA005.7 Preferably, the array of reservoirs has more than 1000 reservoirs.
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GPA005.8 Preferably, the oligonucleotide spotting robot also has CMOS circuitry between the array of reservoirs and the array of ejectors, the CMOS circuitry being configured to drive the array of ejectors in accordance with control signals from the control microprocessor, wherein the CMOS circuitry stores the data relating to the oligonucleotide probes.
GPA005.9 Preferably, the mounting surface is a stage configured for movement along two orthogonal axes, and the array of ejectors is mounted closely adjacent to, and facing, the stage.
GPA005.10 Preferably, the LOC device has an array of hybridization chambers for receiving the oligonucleotide probes, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample and the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs being configured to contain the complete assay of
oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device.
GPA005.11 Preferably, each of the ejectors has a plurality of nozzles, such that the ejector is configured to eject a droplet from each nozzle respectively.
GPA005.12 Preferably, the ejector has a chamber for containing the liquid from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle.
GPA005.13 Preferably, the actuators are thermal actuators, each configured to generate a vapor bubble in the liquid.
GPA005.14 Preferably, the ejectors are configured to eject droplets having a volume less than 100 picoliters.
GPA005.15 Preferably, the ejectors are configured to eject droplets having a volume less than 25 picoliters.
GPA005.16 Preferably, the ejectors are configured to eject droplets having a volume less than 6 picoliters.
GPA005.17 Preferably, the ejectors are configured to eject droplets having a volume between 0.1 picoliters and 1.6 picoliters.
GPA005.18 Preferably, the actuators in each of the ejectors are configured to actuate individually.
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GPA005.19 Preferably, each of the ejectors has a plurality of inlet channels extending from the reservoir to the chamber.
GPA005.20 Preferably, the array of ejectors is configured to spot the oligonucleotides onto the substrate at a rate greater than 100 probe spots per second.
The oligonucleotide spotting robot is used as part of a cost-effective automated mass-manufacturing environment. Loaded oligonucleotide spotting devices are picked up by the robot and the robot positions the oligonucleotide spotting devices and commands them to eject the oligonucleotides into the hybridization chambers of LOC devices that are being spotted. The data automation provided by the oligonucleotide spotting robot includes automated computer-controlled dispensing of the oligonucleotides into the hybridization chambers of LOC devices that are being spotted, reading the specifications of the oligonucleotides stored in the reservoirs of the oligonucleotide spotting devices, checking the specifications of the oligonucleotides stored in the reservoirs of the oligonucleotide spotting devices against the relevant databases, storing the oligonucleotide specifications in the memory of the LOC devices that are being spotted, and transmitting of the
oligonucleotide specifications to other segments of the automated manufacturing environment. The oligonucleotide spotting robot provides for an automated, volumetrically and positionally precise, fast, and high-density oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment. In particular, the capability of the robot to spot at the requisite high-density provides for low final LOC device dimensions, in turn, permitting the inexpensive assay system.
The data automation provided by the oligonucleotide spotting robot provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GSS001.1 This aspect of the invention provides a system for microarray spotting and genetic analysis, the system comprising:
containers with probes for hybridization with different target nucleic acid sequences;
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an integrated circuit secured to each of the containers respectively, each of the integrated circuits have a container digital memory storing probe specification data related to probes in that container;
a micro fluidic device for supporting an array of probes selected from the containers, the selected probes corresponding to a desired genetic test assay, the microfluidic device having a device digital memory;
an oligonucleotide spotting robot for spotting the selected probes onto the microfluidic device to form the array of probes, the oligonucleotide spotting robot having a control microprocessor for downloading the specification data to the device digital memory; and,
a device reader for accessing the specification data from the device digital memory in order to analyze hybridization data from the microfluidic device.
GSS001.2 Preferably, the microfluidic device has:
a supporting substrate;
a microsystems technologies (MST) layer for supporting the array of selected probes; and,
CMOS circuitry between the MST layer and the supporting substrate, the CMOS circuitry having a photosensor for detecting hybridization of probes within the array of selected probes.
GSS001.3 Preferably, the probes are fluorescence resonance energy transfer
(FRET) probes.
GSS001.4 Preferably, the device digital memory also stores device identity data for uniquely identifying the microfluidic device.
GSS001.5 Preferably, the system also has a test module in which the microfluidic device is mounted, the test module being configured for data transmission between the device digital memory and the reader.
GSS001.6 Preferably, the test module connects to the reader via a universal serial bus (USB) connection.
GSS001.7 Preferably, during use, the CMOS circuitry is powered by the reader via the USB connection.
GSS001.8 Preferably, the test module also has an excitation light source for generating a fluorescence emission from the FRET probes which have hybridized with any of the target nucleic acid sequences.
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GSS001.9 Preferably, during use, the excitation light extinguishes prior to activation of the photosensor, and the CMOS circuitry is configured to delay activation of the photosensor for a predetermined period following deactivation of the excitation light source.
GSS001.10 Preferably, the photosensor is less than 249 microns from the probes.
GSS001.1 1 Preferably, the system also has a receptacle for receiving a biological sample containing the target nucleic acid sequences, the receptacle being in fluid communication with an inlet in the microfluidic device.
GSS001.12 Preferably, the microfluidic device has a fluid flow-path from the inlet to the end point sensor, the fluid flow-path configured to bring the target nucleic acid sequences into contact with the array of probes by capillary action.
GSS001.13 Preferably, the microfluidic device has a plurality of reagent reservoirs for different reagents required to process the biological sample.
GSS001.14 Preferably, each of the container digital memories stores identity data distinguishing the container from others used to spot the microfluidic device, the control microprocessor configured to download the identity data to the device digital memory of the microfluidic device being spotted.
GS S001.15 Preferably, the containers each have a droplet generator for ej ecting droplets of a liquid suspension of the probes onto the microfluidic device.
GSS001.16 Preferably, the system also has a mounting surface for mounting the microfluidic device for movement relative to the containers such that the control microprocessor controls both the containers and the mounting section to activate the droplet generator of the container selected and moved into registration with the microfluidic device.
GSS001.17 Preferably, the system also has a camera for optical feedback of the registration between the container selected by the control microprocessor and the microfluidic device.
GSS001.18 Preferably, the data stored in the container digital memory and the device digital memory is encrypted.
GSS001.19 Preferably, the droplet generators are configured to eject droplets having a volume less than 6 picoliters.
GSS001.20 Preferably, the array of selected probes contains more than 1000 probes in an area less than 1500 microns by 1500 microns.
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The system for variable microarray spotting and genetic analysis provides for automated, fast, easy, and low-cost compilation of easy-to-use and inexpensive application-specific/application-optimized assay systems, using a library of LOC devices, an oligonucleotide spotting device, an apparatus for loading of oligonucleotide spotting devices, an oligonucleotide spotting robot, and a library of oligonucleotide probes stored in microvials with digital memory. All the process steps from probe acceptance through LOC device spotting are automated. GSL001.1 This aspect of the invention provides a system for loading reagents into a microfluidic device for genetic analysis, the system comprising:
containers with reagents for processing a biological sample in the microfluidic device;
an integrated circuit secured to each of the containers respectively, each of the integrated circuits having a container digital memory storing reagent specification data related to the reagent in that container;
a microfluidic device for performing a desired genetic test assay, the microfluidic device having a device digital memory;
a reagent dispensing apparatus for loading a selection of the reagents into the microfluidic device, the reagent dispensing apparatus having a control microprocessor for accessing the container digital memory and transmitting the specification data to the device digital memory; and,
a device reader for accessing the reagent specification data from the device digital memory in order to analyze test assay results from the microfluidic device.
GSL001.2 Preferably, the microfluidic device has:
a supporting substrate;
a microsystems technologies (MST) layer for supporting an array of
oligonucleotide probes for hybridization with target nucleic acid sequences in the biological sample; and,
CMOS circuitry between the MST layer and the supporting substrate, the CMOS circuitry having a photosensor for detecting hybridization within the array of probes.
GSL001.3 Preferably, the probes are fluorescence resonance energy transfer (FRET) probes.
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GSL001.4 Preferably, the device digital memory also stores device identity data for uniquely identifying the microfluidic device.
GSL001.5 Preferably, the system also has a test module in which the microfluidic device is mounted, the test module being configured for data transmission between the device digital memory and the reader.
GSL001.6 Preferably, the test module connects to the reader via a universal serial bus (USB) connection.
GSL001.7 Preferably, during use, the CMOS circuitry is powered by the reader via the USB connection.
GSL001.8 Preferably, the test module also has an excitation light source for generating a fluorescence emission from the FRET probes which have hybridized with any of the target nucleic acid sequences.
GSL001.9 Preferably, during use, the excitation light extinguishes prior to activation of the photosensor, and the CMOS circuitry is configured to delay activation the photosensor for a predetermined period following deactivation of the excitation light source.
GSL001.10 Preferably, the photosensor is less than 249 microns from the probes.
GSL001.11 Preferably, the system also has a receptacle for receiving the biological sample containing the target nucleic acid sequences, the receptacle being in fluid communication with an inlet in the microfluidic device.
GSL001.12 Preferably, the microfluidic device has a fluid flow-path from the inlet to the end point sensor, the fluid flow-path configured to bring the target nucleic acid sequences into contact with the array of probes by capillary action.
GSL001.13 Preferably, the microfluidic device has a plurality of reagent reservoirs for the reagents required to process the biological sample.
GS LOO 1.14 Preferably, each of the container digital memories stores identity data distinguishing the container from others used to spot the microfluidic device, the control microprocessor being configured to download the identity data to the device digital memory of the microfluidic device being spotted.
GSL001.15 Preferably, the containers each have a droplet generator for ejecting droplets of the reagent into the reagent reservoirs.
GSL001.16 Preferably, the system also has a mounting surface for mounting the microfluidic device for movement relative to the containers such that the control
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microprocessor controls both the containers and the mounting section to activate the droplet generator of the container selected and moved into registration with the microfluidic device.
GSL001.17 Preferably, the system also has a camera for optical feedback of the registration between the container selected by the control microprocessor and the microfluidic device.
GSL001.18 Preferably, the data stored in the container digital memory and the device digital memory is encrypted.
GSL001.19 Preferably, the droplet generators are configured to eject droplets having a volume less than 6 picoliters.
GSL001.20 Preferably, the array of probes contains more than 1000 probes in an area less than 1500 microns by 1500 microns.
The system for variable LOC device reagent loading and genetic analysis provides for automated, fast, easy, and low-cost compilation of easy-to-use and inexpensive application-specific/application-optimized assay systems, using a library of LOC devices, a reagent dispensing apparatus, and a library of reagents stored in microvials with digital memory. All the process steps from reagent acceptance through LOC device loading are automated.
GCA001.1 This aspect of the invention provides an apparatus for dispensing reagents and loading oligonucleotide spotting devices, the apparatus comprising:
a plurality of reagent vials, each with a droplet dispenser;
a plurality of oligonucleotide vials, each with a droplet dispenser;
a mounting surface for detachably mounting a microfluidic device for movement relative to the reagent vials, and detachably mounting an oligonucleotide spotting device; and,
a control processor for operative control of the reagent vials and oligonucleotide vials and movement of the mounting surface relative to the reagent vials and
oligonucleotide vials; wherein,
the control processor is configured to activate any of the droplet dispensers, move the microfluidic device into registration with the reagent vials and move the
oligonucleotide spotting device into registration with the oligonucleotide vials.
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GCA001.2 Preferably, each of the reagent vials has an integrated circuit storing reagent specification data, each of the oligonucleotide vials has an integrated circuit storing oligonucleotide specification data, and the control processor is configured to download the reagent specification data to digital memory in the microfluidic device and download the oligonucleotide specification data to digital memory in the oligonucleotide spotting device.
GCAOOl .3 Preferably, the apparatus also has a camera for optical feedback of the registration between the vial selected by the control processor and the microfluidic device.
GCAOOl.4 Preferably, the reagent vials and the oligonucleotide vials are microvials with a volume between 282 microliters and 400 microliters.
GCAOOl .5 Preferably, the integrated circuit for each of the microvials has a unique identifier for identifying each of the microvials individually, the unique identifier being transmitted to the control processor.
GCAOOl .6 Preferably, each of the microvials has electrical contacts for receiving activation pulses for the droplet dispenser and allowing the control processor to interrogate the integrated circuit.
GCAOOl.7 Preferably, the apparatus also has a rack wherein the microvials are detachably mounted to the rack for mechanical and electronic control of the microvials.
GCAOOl.8 Preferably, the mounting surface is a stage configured for movement along two orthogonal axes, the rack extending parallel to one if the orthogonal axes.
GCAOOl .9 Preferably, the droplet dispenser has a piezo-electric actuator.
GCAOOl .10 Preferably, the droplet dispenser is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
GCA001.1 1 Preferably, the microfluidic device is a LOC device for genetic analysis of a biological sample, the LOC device having a polymerase chain reaction (PCR) section and the list of reagents has one or more of:
water;
polymerase;
primers;
buffer;
anticoagulant;
deoxyribonucleoside triphosphates (dNTPs);
lysis reagent;
ligase and linkers; and,
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restriction enzymes.
GCA001.12 Preferably, the apparatus also has a facility for applying a film to the LOC device to cover reagent reservoirs formed in an exterior surface.
GCA001.13 Preferably, the LOC device is one of an array of LOC devices fabricated on a silicon wafer, the stage being configured to detachably mount the silicon wafer for loading reagents into all the LOC devices in the array.
GCA001.14 Preferably, the LOC device is one of an array of LOC devices mounted on a printed circuit board (PCB), the stage being configured to detachably mount the PCB for loading reagents into all the LOC devices in the array.
The combined reagent dispensing apparatus and apparatus for loading of oligonucleotide spotting devices is used, as part of a cost-effective automated mass- manufacturing environment, to dispense reagents contained in reagent microvials into the reagent reservoirs of microfluidic devices and to dispense oligonucleotides contained in oligonucleotide microvials into the oligonucleotide reservoirs of oligonucleotide spotting devices. The data automation provided by the apparatus includes automated computer- controlled dispensing of the reagents into the reagent reservoirs of the microfluidic devices and dispensing of the oligonucleotides into the oligonucleotide reservoirs of the oligonucleotide spotting devices, checking the reagent data stored in the memory of the microvials against the list of specifications for the reagents that have to be loaded in the microfluidic device, checking the oligonucleotide data stored in the memory of the microvials against the list of specifications for the oligonucleotides that have to be loaded in the oligonucleotide spotting devices, storage of the reagent data into the memory of the microfluidic device, and storage of the oligonucleotide data into the memory of the oligonucleotide spotting devices.
The combined reagent dispensing apparatus and apparatus for loading of oligonucleotide spotting devices provides for an automated and volumetrically and positionally precise reagent and oligonucleotide dispensing technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
The data automation provided by the combined reagent dispensing apparatus and apparatus for loading of oligonucleotide spotting devices provides for an automated, safe,
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secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GCA002.1 This aspect of the invention provides an apparatus for dispensing reagents, loading oligonucleotide spotting devices and spotting oligonucleotide probes, the apparatus comprising:
a plurality of reagent vials, each with a droplet dispenser;
a plurality of oligonucleotide vials, each with a droplet dispenser;
a mounting surface for detachably mounting a microfluidic device for movement relative to the reagent vials, and detachably mounting an oligonucleotide spotting device; a chuck for detachably mounting the oligonucleotide spotting device adjacent the mounting surface; and,
a control processor for operative control of the reagent and oligonucleotide vials, the oligonucleotide spotting device when mounted in the chuck and movement of the mounting surface relative to the reagent and oligonucleotide vials, and the oligonucleotide spotting device; wherein,
the control processor is configured to activate any of the droplet dispensers, move the microfluidic device into registration with the reagent vials and move the
oligonucleotide spotting device into registration with the oligonucleotide vials.
GCA002.2 Preferably, the control processor is configured to operate the oligonucleotide spotting device when in the chuck to spot oligonucleotide probes into the microfluidic device on the mounting surface.
GCA002.3 Preferably, each of the reagent vials has an integrated circuit storing reagent specification data, each of the oligonucleotide vials has an integrated circuit storing oligonucleotide specification data, and the control processor is configured to download the reagent specification data to digital memory in the microfluidic device and download the oligonucleotide specification data to digital memory in the oligonucleotide spotting device.
GCA002.4 Preferably, the apparatus also has a camera for optical feedback of the registration between the vial selected by the control processor and the microfluidic device.
GCA002.5 Preferably, the reagent vials and the oligonucleotide vials are microvials with a volume between 282 microliters and 400 microliters.
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GCA002.6 Preferably, the integrated circuit for each of the micro vials has a unique identifier for identifying each of the micro vials individually, the unique identifier being transmitted to the control processor.
GCA002.7 Preferably, each of the microvials has electrical contacts for receiving activation pulses for the droplet dispenser and allowing the control processor to interrogate the integrated circuit.
GCA002.8 Preferably, the apparatus also has a rack wherein the microvials are detachably mounted to the rack for mechanical and electronic control of the microvials.
GCA002.9 Preferably, the mounting surface is a stage configured for movement along two orthogonal axes, the rack extending parallel to one if the orthogonal axes.
GCA002.10 Preferably, the droplet dispenser has a piezo-electric actuator.
GCA002.1 1 Preferably, the droplet dispenser is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
GCA002.12 Preferably, the micro fluidic device is a LOC device for genetic analysis of a biological sample, the LOC device having a polymerase chain reaction (PCR) section and the list of reagents has one or more of:
water;
polymerase;
primers;
buffer;
anticoagulant;
deoxyribonucleoside triphosphates (dNTPs);
lysis reagent;
ligase and linkers; and,
restriction enzymes.
GCA002.13 Preferably, the apparatus also has a facility for applying a film to the LOC device to cover reagent reservoirs formed in an exterior surface.
GCA002.14 Preferably, the LOC device is one of an array of LOC devices fabricated on a silicon wafer, the stage being configured to detachably mount the silicon wafer for loading reagents into all the LOC devices in the array.
GCA002.15 Preferably, the LOC device is one of an array of LOC devices mounted on a printed circuit board (PCB), the stage being configured to detachably mount the PCB for loading reagents into all the LOC devices in the array.
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GCA002.16 Preferably, the oligonucleotide spotting device has an array of reservoirs for containing the oligonucleotide probes and an array of ejectors, and the LOC device has an array of hybridization chambers for receiving the oligonucleotide probes, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample and the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device, the control processor being configured to operate the ejectors to correctly spot the hybridization chamber array and download an association between the specification data for the oligonucleotide probes from each of the reservoirs, and array location data locating the hybridization chamber spotted by each of the reservoirs.
GCA002.17 Preferably, each of the ejectors has a plurality of nozzles, a chamber for containing liquid with a suspension of the oligonucleotide probes from the
corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle, the control processor being configured to operate each of the actuators individually.
GCA002.18 Preferably, the control processor is configured to operate the array of ejectors to spot the oligonucleotides onto the LOC device with a density greater than 8 probe spots per square millimeter.
GCA002.19 Preferably, the control processor is configured to operate the array of ejectors to spot the oligonucleotides onto the LOC device with a density greater than 60 probe spots per square millimeter.
GCA002.20 Preferably, the control processor is configured to operate the array of ejectors to spot the oligonucleotides onto the LOC device with a density between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
The combined apparatus for reagent dispensing, loading of oligonucleotide spotting devices, and oligonucleotide spotting is used, as part of a cost-effective automated mass- manufacturing environment, to dispense reagents contained in reagent microvials into the reagent reservoirs of microfluidic devices, to dispense oligonucleotides contained in oligonucleotide microvials into the oligonucleotide reservoirs of oligonucleotide spotting
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devices, and to eject the oligonucleotides onto the surfaces that are being spotted. The data automation provided by the apparatus includes automated computer-controlled dispensing of the reagents into the reagent reservoirs of the microfluidic devices, dispensing of the oligonucleotides into the oligonucleotide reservoirs of the oligonucleotide spotting devices, dispensing of the oligonucleotides onto the surface being spotted, checking the reagent data stored in the memory of the microvials against the list of specifications for the reagents that have to be loaded in the microfluidic device, checking the oligonucleotide data stored in the memory of the microvials against the list of specifications for the oligonucleotides that have to be loaded in the oligonucleotide spotting devices, storing the reagent data into the memory of the microfluidic device, storing the oligonucleotide specifications in the memory of the devices that are being spotted, and transmitting of the reagent and oligonucleotide specifications to other segments of the automated
manufacturing environment. The combined apparatus for reagent dispensing, loading of oligonucleotide spotting devices, and oligonucleotide spotting provides for an automated, volumetrically and positionally precise, fast, and high-density reagent dispensing and oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
The data automation provided by the combined apparatus for reagent dispensing, loading of oligonucleotide spotting devices, and oligonucleotide spotting provides for an automated, safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
GCA003.1 This aspect of the invention provides an apparatus for loading oligonucleotide spotting devices and spotting oligonucleotide probes, the apparatus comprising:
a plurality of oligonucleotide vials, each with a droplet dispenser;
a mounting surface for detachably mounting an oligonucleotide spotting device; a chuck for detachably mounting the oligonucleotide spotting device adjacent the mounting surface; and,
a control processor for operative control of the oligonucleotide vials, the oligonucleotide spotting device when mounted in the chuck and movement of the
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mounting surface relative to the oligonucleotide vials, and the oligonucleotide spotting device; wherein,
the control processor is configured to activate the droplet dispensers, and move the oligonucleotide spotting device into registration with the oligonucleotide vials.
GCA003.2 Preferably, the control processor is configured to operate the oligonucleotide spotting device when in the chuck to spot oligonucleotide probes into a microfluidic device on the mounting surface.
GCA003.3 Preferably, each of the oligonucleotide vials has an integrated circuit storing oligonucleotide specification data, and the control processor is configured to download the oligonucleotide specification data to digital memory in the oligonucleotide spotting device.
GCA003.4 Preferably, the apparatus also has a camera for optical feedback of the registration between the vial selected by the control processor and the oligonucleotide spotting device.
GCA003.5 Preferably, the oligonucleotide vials are micro vials with a volume between 282 microliters and 400 microliters.
GCA003.6 Preferably, the integrated circuit for each of the micro vials has a unique identifier for identifying each of the micro vials individually, the unique identifier being transmitted to the control processor.
GCA003.7 Preferably, each of the microvials has electrical contacts for receiving activation pulses for the droplet dispenser and allowing the control processor to interrogate the integrated circuit.
GCA003.8 Preferably, the apparatus also has a rack wherein the microvials are detachably mounted to the rack for mechanical and electronic control of the microvials.
GCA003.9 Preferably, the mounting surface is a stage configured for movement along two orthogonal axes, the rack extending parallel to one if the orthogonal axes.
GCA003.10 Preferably, the droplet dispenser has a piezo-electric actuator.
GCA003.1 1 Preferably, the droplet dispenser is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
GCA003.12 Preferably, the apparatus also has reagent vials containing reagents for processing a biological sample wherein the microfluidic device is a LOC device for genetic analysis of the biological sample, the LOC device having a polymerase chain reaction (PC ) section and the list of reagents has one or more of:
water;
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polymerase;
primers;
buffer;
anticoagulant;
deoxyribonucleoside triphosphates (dNTPs);
lysis reagent;
ligase and linkers; and,
restriction enzymes.
GCA003.13 Preferably, the apparatus also has a facility for applying a film to the LOC device to cover reagent reservoirs formed in an exterior surface.
GCA003.14 Preferably, the LOC device is one of an array of LOC devices fabricated on a silicon wafer, the stage being configured to detachably mount the silicon wafer for loading reagents into all the LOC devices in the array.
GCA003.15 Preferably, the LOC device is one of an array of LOC devices mounted on a printed circuit board (PCB), the stage being configured to detachably mount the PCB for loading reagents into all the LOC devices in the array.
GCA003.16 Preferably, the oligonucleotide spotting device has an array of reservoirs for containing the oligonucleotide probes and an array of ejectors, and the LOC device has an array of hybridization chambers for receiving the oligonucleotide probes, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample and the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device, the control processor being configured to operate the ejectors to correctly spot the hybridization chamber array and download an association between the specification data for the oligonucleotide probes from each of the reservoirs, and array location data locating the hybridization chamber spotted by each of the reservoirs.
GCA003.17 Preferably, each of the ejectors has a plurality of nozzles, a chamber for containing liquid with a suspension of the oligonucleotide probes from the
corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the
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chamber through the corresponding nozzle, the control processor being configured to operate each of the actuators individually.
GCA003.18 Preferably, the control processor is configured to operate the array of ejectors to spot the oligonucleotides onto the LOC device with a density greater than 8 probe spots per square millimeter.
GCA003.19 Preferably, the control processor is configured to operate the array of ejectors to spot the oligonucleotides onto the LOC device with a density greater than 60 probe spots per square millimeter.
GCA003.20 Preferably, the control processor is configured to operate the array of ejectors to spot the oligonucleotides onto the LOC device with a density between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
The combined apparatus for loading of oligonucleotide spotting devices and oligonucleotide spotting is used, as part of a cost-effective automated mass-manufacturing environment, to dispense oligonucleotides contained in oligonucleotide microvials into the oligonucleotide reservoirs of oligonucleotide spotting devices and to eject the
oligonucleotides onto the surfaces that are being spotted. The data automation provided by the apparatus includes automated computer-controlled dispensing of the oligonucleotides into the oligonucleotide reservoirs of the oligonucleotide spotting devices, dispensing of the oligonucleotides onto the surface being spotted, checking the oligonucleotide data stored in the memory of the microvials against the list of specifications for the oligonucleotides that have to be loaded in the oligonucleotide spotting devices, storing the oligonucleotide specifications in the memory of the devices that are being spotted, and transmitting of the oligonucleotide specifications to other segments of the automated manufacturing environment.
The combined apparatus for loading of oligonucleotide spotting devices and oligonucleotide spotting provides for an automated, volumetrically and positionally precise, fast, and high-density and oligonucleotide spotting technique, simplifying the complexity, increasing the reliability, increasing the security, increasing the safety, and reducing the cost of the automated manufacturing environment.
The data automation provided by the combined apparatus for loading of oligonucleotide spotting devices and oligonucleotide spotting provides for an automated,
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safe, secure, and inexpensive technique of data monitoring and management in the automated manufacturing environment.
BRIEF DESCRIPTION OF THE DRAWINGS
Preferred embodiments of the present invention will now be described by way of example only with reference to the accompanying drawings, in which:
Figure 1 shows a test module and test module reader configured for fluorescence detection;
Figure 2 is a schematic overview of the electronic components in the test module configured for fluorescence detection;
Figure 3 is a schematic overview of the electronic components in the test module reader;
Figure 4 is a schematic representation of the architecture of the LOC device; Figure 5 is a perspective of the LOC device;
Figure 6 is a plan view of the LOC device with features and structures from all layers superimposed on each other;
Figure 7 is a plan view of the LOC device with the structures of the cap shown in isolation;
Figure 8 is a top perspective of the cap with internal channels and reservoirs shown in dotted line;
Figure 9 is an exploded top perspective of the cap with internal channels and reservoirs shown in dotted line;
Figure 10 is a bottom perspective of the cap showing the configuration of the top channels;
Figure 11 is a plan view of the LOC device showing the structures of the CMOS + MST device in isolation;
Figure 12 is a schematic section view of the LOC device at the sample inlet;
Figure 13 is an enlarged view of Inset AA shown in Figure 6;
Figure 14 is an enlarged view of Inset AB shown in Figure 6;
Figure 15 is an enlarged view of Inset AE shown in Figure 13;
Figure 16 is a partial perspective illustrating the laminar structure of the LOC device within Inset AE;
Figure 17 is a partial perspective illustrating the laminar structure of the LOC device within Inset AE;
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Figure 18 is a partial perspective illustrating the laminar structure of the LOC device within Inset AE;
Figure 19 is a partial perspective illustrating the laminar structure of the LOC device within Inset AE;
Figure 20 is a partial perspective illustrating the laminar structure of the LOC device within Inset AE;
Figure 21 is a partial perspective illustrating the laminar structure of the LOC device within Inset AE;
Figure 22 is schematic section view of the lysis reagent reservoir shown in Figure 21;
Figure 23 is a partial perspective illustrating the laminar structure of the LOC device within Inset AB;
Figure 24 is a partial perspective illustrating the laminar structure of the LOC device within Inset AB;
Figure 25 is a partial perspective illustrating the laminar structure of the LOC device within Inset AI;
Figure 26 is a partial perspective illustrating the laminar structure of the LOC device within Inset AB;
Figure 27 is a partial perspective illustrating the laminar structure of the LOC device within Inset AB;
Figure 28 is a partial perspective illustrating the laminar structure of the LOC device within Inset AB;
Figure 29 is a partial perspective illustrating the laminar structure of the LOC device within Inset AB;
Figure 30 is a schematic section view of the amplification mix reservoir and the polymerase reservoir;
Figure 31 show the features of a boiling- initiated valve in isolation;
Figure 32 is a schematic section view of the boiling-initiated valve taken through line 33-33 shown in Figure 31;
Figure 33 is an enlarged view of Inset AF shown in Figure 15;
Figure 34 is a schematic section view of the upstream end of the dialysis section taken through line 35-35 shown in Figure 33;
Figure 35 is an enlarged view of Inset AC shown in Figure 6;
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Figure 36 is a further enlarged view within Inset AC showing the amplification section;
Figure 37 is a further enlarged view within Inset AC showing the amplification section;
Figure 38 is a further enlarged view within Inset AC showing the amplification section;
Figure 39 is a further enlarged view within Inset AK shown in Figure 38; Figure 40 is a further enlarged view within Inset AC showing the amplification chamber;
Figure 41 is a further enlarged view within Inset AC showing the amplification section;
Figure 42 is a further enlarged view within Inset AC showing the amplification chamber;
Figure 43 is a further enlarged view within Inset AL shown in Figure 42; Figure 44 is a further enlarged view within Inset AC showing the amplification section;
Figure 45 is a further enlarged view within Inset AM shown in Figure 44; Figure 46 is a further enlarged view within Inset AC showing the amplification chamber;
Figure 47 is a further enlarged view within Inset AN shown in Figure 46;
Figure 48 is a further enlarged view within Inset AC showing the amplification chamber;
Figure 49 is a further enlarged view within Inset AC showing the amplification chamber;
Figure 50 is a further enlarged view within Inset AC showing the amplification section;
Figure 51 is a schematic section view of the amplification section;
Figure 52 is an enlarged plan view of the hybridization section;
Figure 53 is a further enlarged plan view of two hybridization chambers in isolation;
Figure 54 is schematic section view of a single hybridization chamber;
Figure 55 is an enlarged view of the humidifier illustrated in Inset AG shown in Figure 6;
Figure 56 is an enlarged view of Inset AD shown in Figure 52;
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Figure 57 is an exploded perspective view of the LOC device within Inset AD; Figure 58 is a diagram of a FRET probe in a closed configuration;
Figure 59 is a diagram of a FRET probe in an open and hybridized configuration; Figure 60 is a graph of the intensity of an excitation light over time;
Figure 61 is a diagram of the excitation illumination geometry of the hybridization chamber array;
Figure 62 is a diagram of a Sensor Electronic Technology LED illumination geometry;
Figure 63 is a schematic plan view of a reagent dispensing robot;
Figure 64 is a perspective of a reagent microvial with inbuilt droplet generator;
Figure 65 is a schematic plan view of an oligonucleotide ejector robot for loading selected probes into a probe ejector chip;
Figure 66 is a schematic plan view of a probe spotting robot for loading probes into the LOC devices on a partial-depth sawn silicon wafer;
Figure 67 is an enlarged plan view of the humidity sensor shown in Inset AH of Figure 6;
Figure 68 is a schematic showing part of the photodiode array of the photosensor;
Figure 69 is a circuit diagram for a single photodiode;
Figure 70 is a timing diagram for the photodiode control signals;
Figure 71 shows an oligonucleotide ejector chip (ONEC);
Figure 72 shows an array of droplet generators from the ONEC shown in Inset AO of Figure 71 ;
Figure 73 is a schematic section of the array of droplet generators taken along line 91-91 shown in Figure 72;
Figure 74 is an enlarged view of the evaporator shown in Inset AP of Figure 55;
Figure 75 is a schematic section view through a hybridization chamber with a detection photodiode and trigger photodiode;
Figure 76 is a diagram of linker-primed PCR;
Figure 77 is a schematic representation of a test module with a lancet;
Figure 78 is a diagrammatic representation of the architecture of LOC variant VII; Figure 79 is a diagrammatic representation of the architecture of LOC variant VIII; Figure 80 is a schematic illustration of the architecture of LOC variant XIV;
Figure 81 is a schematic illustration of the architecture of LOC variant XLI;
Figure 82 is a schematic illustration of the architecture of LOC variant XLIII;
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Figure 83 is a schematic illustration of the architecture of LOC variant XLIV;
Figure 84 is a schematic illustration of the architecture of LOC variant XLVII;
Figure 85 is a diagram of a primer-linked, linear fluorescent probe during the initial round of amplification;
Figure 86 is a diagram of a primer-linked, linear fluorescent probe during a subsequent amplification cycle;
Figures 87A to 87F diagrammatically illustrate thermal cycling of a primer-linked fluorescent stem-and-loop probe;
Figure 88 is a schematic illustration of the excitation LED relative to the hybridization chamber array and the photodiodes;
Figure 89 is a schematic illustration of the excitation LED and optical lens for directing light onto the hybridization chamber array of the LOC device;
Figure 90 is a schematic illustration of the excitation LED, optical lens, and optical prisms for directing light onto the hybridization chamber array of the LOC device;
Figure 91 is a schematic illustration of the excitation LED, optical lens and mirror arrangement for directing light onto the hybridization chamber array of the LOC device;
Figure 92 is a schematic plan view of a probe spotting robot for loading probes into the LOC devices on a separable PCB;
Figure 93 is a plan view showing all the features superimposed on each other, and showing the location of Insets DA to DK;
Figure 94 is an enlarged view of Inset DG shown in Figure 93;
Figure 95 is an enlarged view of Inset DH shown in Figure 93;
Figure 96 shows one embodiment of the shunt transistor for the photodiodes;
Figure 97 shows one embodiment of the shunt transistor for the photodiodes;
Figure 98 shows one embodiment of the shunt transistor for the photodiodes;
Figure 99 is a circuit diagram of the differential imager;
Figure 100 schematically illustrates a negative control fluorescent probe in its stem- and-loop configuration;
Figure 101 schematically illustrates the negative control fluorescent probe of Figure 100 in its open configuration;
Figure 102 schematically illustrates a positive control fluorescent probe in its stem- and-loop configuration;
Figure 103 schematically illustrates the positive control fluorescent probe of Figure 102 in its open configuration;
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Figure 104 shows a test module and test module reader configured for use with ECL detection;
Figure 105 is a schematic overview of the electronic components in the test module configured for use with ECL detection;
Figure 106 shows a test module and alternative test module readers;
Figure 107 shows a test module and test module reader along with the hosting system housing various databases;
Figure 108 is a schematic side view of a reagent spotting robot;
Figure 109 is a schematic representation of an electrochemiluminescence-based test module with multidevice micro fluidic device;
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
OVERVIEW
This overview identifies the main components of a molecular diagnostic system that incorporates embodiments of the present invention. Comprehensive details of the system architecture and operation are set out later in the specification.
Referring to Figures 1, 2, 3, 104 and 105, the system has the following top level components:
Test modules 10 and 1 1 are the size of a typical USB memory key and very cheap to produce. Test modules 10 and 11 each contain a microfluidic device, typically in the form of a lab-on-a-chip (LOC) device 30 preloaded with reagents and typically more than 1000 probes for the molecular diagnostic assay (see Figures 1 and 104). Test module 10 schematically shown in Figure 1 uses a fluorescence-based detection technique to identify target molecules, while test module 1 1 in Figure 104 uses an electrochemiluminescence- based detection technique. The LOC device 30 has an integrated photosensor 44 for fluorescence or electrochemiluminescence detection (described in detail below). Both test modules 10 and 11 use a standard Micro-USB plug 14 for power, data and control, both have a printed circuit board (PCB) 57, and both have external power supply capacitors 32 and an inductor 15. The test modules 10 and 11 are both single-use only for mass production and distribution in sterile packaging ready for use.
The outer casing 13 has a macroreceptacle 24 for receiving the biological sample and a removable sterile sealing tape 22, preferably with a low tack adhesive, to cover the macroreceptacle prior to use. A membrane seal 408 with a membrane guard 410 forms
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part of the outer casing 13 to reduce dehumidification within the test module while providing pressure relief from small air pressure fluctuations. The membrane guard 410 protects the membrane seal 408 from damage.
Test module reader 12 powers the test module 10 or 11 via Micro-USB port 16. The test module reader 12 can adopt many different forms and a selection of these are described later. The version of the reader 12 shown in Figures 1, 3 and 104 is a smart phone embodiment. A block diagram of this reader 12 is shown in Figure 3. Processor 42 runs application software from program storage 43. The processor 42 also interfaces with the display screen 18 and user interface (UI) touch screen 17 and buttons 19, a cellular radio 21, wireless network connection 23, and a satellite navigation system 25. The cellular radio 21 and wireless network connection 23 are used for communications.
Satellite navigation system 25 is used for updating epidemiological databases with location data. The location data can, alternatively, be entered manually via the touch screen 17 or buttons 19. Data storage 27 holds genetic and diagnostic information, test results, patient information, assay and probe data for identifying each probe and its array position. Data storage 27 and program storage 43 may be shared in a common memory facility.
Application software installed on the test module reader 12 provides analysis of results, along with additional test and diagnostic information.
To conduct a diagnostic test, the test module 10 (or test module 1 1) is inserted into the Micro-USB port 16 on the test module reader 12. The sterile sealing tape 22 is peeled back and the biological sample (in a liquid form) is loaded into the sample macroreceptacle 24. Pressing start button 20 initiates testing via the application software. The sample flows into the LOC device 30 and the on-board assay extracts, incubates, amplifies and hybridizes the sample nucleic acids (the target) with presynthesized hybridization- responsive oligonucleotide probes. In the case of test module 10 (which uses fluorescence- based detection), the probes are fluorescently labelled and the LED 26 housed in the casing 13 provides the necessary excitation light to induce fluorescence emission from the hybridized probes (see Figures 1 and 2). In test module 11 (which uses
electrochemiluminescence (ECL) detection), the LOC device 30 is loaded with ECL probes (discussed above) and the LED 26 is not necessary for generating the luminescent emission. Instead, electrodes 860 and 870 provide the excitation electrical current (see Figure 105). The emission (fluorescent or luminescent) is detected using a photosensor 44 integrated into CMOS circuitry of each LOC device. The detected signal is amplified and
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converted to a digital output which is analyzed by the test module reader 12. The reader then displays the results.
The data may be saved locally and'or uploaded to a network server containing patient records. The test module 10 or 11 is removed from the test module reader 12 and disposed of appropriately.
Figures 1, 3 and 104 show the test module reader 12 configured as a mobile phone/smart phone 28. In other forms, the test module reader is a laptop/notebook 101, a dedicated reader 103, an ebook reader 107, a tablet computer 109 or desktop computer 105 for use in hospitals, private practices or laboratories (see Figure 106). The reader can interface with a range of additional applications such as patient records, billing, online databases and multi-user environments. It can also be interfaced with a range of local or remote peripherals such as printers and patient smart cards.
Referring to Figure 107, the data generated by the test module 10 can be used to update, via the reader 12 and network 125, the epidemiological databases hosted on the hosting system for epidemiological data 11 1, the genetic databases hosted on the hosting system for genetic data 113, the electronic health records hosted on the hosting system for electronic health records (EHR) 1 15, the electronic medical records hosted on the hosting system for electronic medical records (EMR) 121, and the personal health records hosted on the hosting system for personal health records (PHR) 123. Conversely, the epidemiological data hosted on the hosting system for epidemiological data 111 , the genetic data hosted on the hosting system for genetic data 113, the electronic health records hosted on the hosting system for electronic health records (EHR) 1 15, the electronic medical records hosted on the hosting system for electronic medical records (EMR) 121, and the personal health records hosted on the hosting system for personal health records (PHR) 123, can be used to update, via network 125 and the reader 12, the digital memory in the LOC 30 of the test module 10.
Referring back to Figures 1, 2, 104 and 105 the reader 12 uses battery power in the mobile phone configuration. The mobile phone reader contains all test and diagnostic information preloaded. Data can also be loaded or updated via a number of wireless or contact interfaces to enable communications with peripheral devices, computers or online servers. A Micro-USB port 16 is provided for connection to a computer or mains power supply for battery recharge.
Figure 77 shows an embodiment of the test module 10 used for tests that only require a positive or negative result for a particular target, such as testing whether a person
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is infected with, for example, H1N1 Influenza A virus. Only a purpose built USB power/indicator-only module 47 is adequate. No other reader or application software is necessary. An indicator 45 on the USB power/indicator-only module 47 signals positive or negative results. This configuration is well suited to mass screening.
Additional items supplied with the system may include a test tube containing reagents for pre-treatment of certain samples, along with spatula and lancet for sample collection. Figure 77 shows an embodiment of the test module incorporating a spring- loaded, retractable lancet 390 and lancet release button 392 for convenience. A satellite phone can be used in remote areas. TEST MODULE ELECTRONICS
Figures 2 and 105 are block diagrams of the electronic components in the test modules 10 and 11, respectively. The CMOS circuitry integrated in the LOC device 30 has a USB device driver 36, a controller 34, a USB-compatible LED driver 29, clock 33, power conditioner 31, RAM 38 and program and data flash memory 40. These provide the control and memory for the entire test module 10 or 11 including the photosensor 44, the temperature sensors 170, the liquid sensors 174, and the various heaters 152, 154, 182, 234, together with associated drivers 37 and 39 and registers 35 and 41. Only the LED 26 (in the case of test module 10), external power supply capacitors 32 and the Micro-USB plug 14 are external to the LOC device 30. The LOC devices 30 include bond-pads for making connections to these external components. The RAM 38 and the program and data flash memory 40 have the application software and the diagnostic and test information (Flash/Secure storage, e.g. via encryption) for over 1000 probes. In the case of test module 1 1 configured for ECL detection, there is no LED 26 (see Figures 104 and 105). Data is encrypted by the LOC device 30 for secure storage and secure communication with an external device. The LOC devices 30 are loaded with electrochemiluminescent probes and the hybridization chambers each have a pair of ECL excitation electrodes 860 and 870.
Many types of test modules 10 are manufactured in a number of test forms, ready for off-the-shelf use. The differences between the test forms lie in the on board assay of reagents and probes.
Some examples of infectious diseases rapidly identified with this system include:
• Influenza - Influenza virus A, B, C, Isavirus, Thogotovirus
• Pneumonia - respiratory syncytial virus (RSV), adenovirus, metapneumovirus,
Streptococcus pneumoniae, Staphylococcus aureus
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• Tuberculosis - Mycobacterium tuberculosis, bovis, africanum, canetti, and microti
• Plasmodium falciparum, Toxoplasma gondii and other protozoan parasites
• Typhoid - Salmonella enterica serovar typhi
• Ebola virus
• Human immunodeficiency virus (HIV)
• Dengue Fever - Flavivirus
• Hepatitis (A through E)
• Hospital acquired infections - for example Clostridium difficile, Vancomycin resistant Enterococcus, and Methicillin resistant Staphylococcus aureus
• Herpes simplex virus (HSV)
• Cytomegalovirus (CMV)
• Epstein-Barr virus (EBV)
• Encephalitis - Japanese Encephalitis virus, Chandipura virus
• Whooping cough - Bordetella pertussis
• Measles - paramyxovirus
• Meningitis - Streptococcus pneumoniae and Neisseria meningitidis
• Anthrax - Bacillus anthracis
Some examples of genetic disorders identified with this system include:
Cystic fibrosis
Haemophilia
Sickle cell disease
Tay-Sachs disease
Haemochromatosis
Cerebral arteriopathy
Crohn's disease
Polycistic kidney disease
Congential heart disease
Rett syndrome
A small selection of cancers identified by the diagnostic system include:
• Ovarian
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• Colon carcinoma
• Multiple endocrine neoplasia
• Retinoblastoma
• Turcot syndrome
The above lists are not exhaustive and the diagnostic system can be configured to detect a much greater variety of diseases and conditions using nucleic acid and proteomic analysis.
DETAILED ARCHITECTURE OF SYSTEM COMPONENTS LOC DEVICE
The LOC device 30 is central to the diagnostic system. It rapidly performs the four major steps of a nucleic acid based molecular diagnostic assay, i.e. sample preparation, nucleic acid extraction, nucleic acid amplification, and detection, using a microfluidic platform. The LOC device also has alternative uses, and these are detailed later. As discussed above, test modules 10 and 11 can adopt many different configurations to detect different targets. Likewise, the LOC device 30 has numerous different embodiments tailored to the target(s) of interest. One form of the LOC device 30 is LOC device 301 for fluorescent detection of target nucleic acid sequences in the pathogens of a whole blood sample. For the purposes of illustration, the structure and operation of LOC device 301 is now described in detail with reference to Figures 4 to 26 and 27 to 57.
Figure 4 is a schematic representation of the architecture of the LOC device 301. For convenience, process stages shown in Figure 4 are indicated with the reference numeral corresponding to the functional sections of the LOC device 301 that perform that process stage. The process stages associated with each of the major steps of a nucleic acid based molecular diagnostic assay are also indicated: sample input and preparation 288, extraction 290, incubation 291, amplification 292 and detection 294. The various reservoirs, chambers, valves and other components of the LOC device 301 will be described in more detail later.
Figure 5 is a perspective view of the LOC device 301. It is fabricated using high volume CMOS and MST (microsystems technology) manufacturing techniques. The laminar structure of the LOC device 301 is illustrated in the schematic (not to scale) partial section view of Figure 12. The LOC device 301 has a silicon substrate 84 which supports
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the CMOS + MST chip 48, comprising CMOS circuitry 86 and an MST layer 87, with a cap 46 overlaying the MST layer 87. For the purposes of this patent specification, the term 'MST layer' is a reference to a collection of structures and layers that process the sample with various reagents. Accordingly, these structures and components are configured to define flow-paths with characteristic dimensions that will support capillary driven flow of liquids with physical characteristics similar to those of the sample during processing. In light of this, the MST layer and components are typically fabricated using surface micromachining techniques and/or bulk micromachining techniques. However, other fabrication methods can also produce structures and components dimensioned for capillary driven flows and processing very small volumes. The specific embodiments described in this specification show the MST layer as the structures and active components supported on the CMOS circuitry 86, but excluding the features of the cap 46. However, the skilled addressee will appreciate that the MST layer need not have underlying CMOS or indeed an overlying cap in order for it to process the sample.
The overall dimensions of the LOC device shown in the following figures are
1760μιη x 824μιη. Of course, LOC devices fabricated for different applications may have different dimensions.
Figure 6 shows the features of the MST layer 87 superimposed with the features of the cap. Insets AA to AD, AG and AH shown in Figure 6 are enlarged in Figures 13, 14, 3 , 56, 55 and 67, respectively, and described in detail below for a comprehensive understanding of each structure within the LOC device 301. Figures 7 to 10 show the features of the cap 46 in isolation while Figure 1 1 shows the CMOS + MST device 48 structures in isolation.
LAMINAR STRUCTURE Figures 12 and 22 are sketches that diagrammatically show the laminar structure of the CMOS + MST device 48, the cap 46 and the fluidic interaction between the two. The figures are not to scale for the purposes of illustration. Figure 12 is a schematic section view through the sample inlet 68 and Figure 22 is a schematic section through the reservoir 54. As best shown in Figure 12, the CMOS + MST device 48 has a silicon substrate 84 which supports the CMOS circuitry 86 that operates the active elements within the MST layer 87 above. A passivation layer 88 seals and protects the CMOS layer 86 from the fluid flows through the MST layer 87.
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Fluid flows through both the cap channels 94 and the MST channels 90 (see for example Figures 7 and 16) in the cap layer 46 and MST channel layer 100, respectively. Cell transport occurs in the larger channels 94 fabricated in the cap 46, while biochemical processes are carried out in the smaller MST channels 90. Cell transport channels are sized so as to be able to transport cells in the sample to predetermined sites in the MST channels 90. Transportation of cells with sizes greater than 20 microns (for example, certain leukocytes) requires channel dimensions greater than 20 microns, and therefore a cross sectional area transverse to the flow of greater than 400 square microns. MST channels, particularly at locations in the LOC where transport of cells is not required, can be significantly smaller.
It will be appreciated that cap channel 94 and MST channel 90 are generic references and particular MST channels 90 may also be referred to as (for example) heated microchannels or dialysis MST channels in light of their particular function. MST channels 90 are formed by etching through a MST channel layer 100 deposited on the passivation layer 88 and patterned with photoresist. The MST channels 90 are enclosed by a roof layer 66 which forms the top (with respect to the orientation shown in the figures) of the CMOS + MST device 48.
Despite sometimes being shown as separate layers, the cap channel layer 80 and the reservoir layer 78 are formed from a unitary piece of material. Of course, the piece of material may also be non-unitary. This piece of material is etched from both sides in order to form a cap channel layer 80 in which the cap channels 94 are etched and the reservoir layer 78 in which the reservoirs 54, 56, 58, 60 and 62 are etched. Alternatively, the reservoirs and the cap channels are formed by a micromolding process. Both etching and micromolding techniques are used to produce channels with cross sectional areas transverse to the flow as large as 20,000 square microns, and as small as 8 square microns.
At different locations in the LOC device, there can be a range of appropriate choices for the cross sectional area of the channel transverse to the flow. Where large quantities of sample, or samples with large constituents, are contained in the channel, a cross-sectional area of up to 20,000 square microns (for example, a 200 micron wide channel in a 100 micron thick layer) is suitable. Where small quantities of liquid, or mixtures without large cells present, are contained in the channel, a very small cross sectional area transverse to the flow is preferable.
A lower seal 64 encloses the cap channels 94 and the upper seal layer 82 encloses the reservoirs 54, 56, 58, 60 and 62.
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The five reservoirs 54, 56, 58, 60 and 62 are preloaded with assay-specific reagents. In the embodiment described here, the reservoirs are preloaded with the following reagents, but other reagents can easily be substituted: · reservoir 54: anticoagulant with option to include erythrocyte lysis buffer
• reservoir 56: lysis reagent
• reservoir 58: restriction enzymes, ligase and linkers (for linker-primed PCR (see Figure 76, extracted from T. Stachan et al, Human Molecular Genetics 2, Garland Science, NY and London, 1999))
· reservoir 60: amplification mix (dNTPs, primers, buffer) and
• reservoir 62: DNA polymerase.
The cap 46 and the CMOS+MST layers 48 are in fluid communication via corresponding openings in the lower seal 64 and the roof layer 66. These openings are referred to as uptakes 96 and downtakes 92 depending on whether fluid is flowing from the MST channels 90 to the cap channels 94 or vice versa.
LOC DEVICE OPERATION
The operation of the LOC device 301 is described below in a step-wise fashion with reference to analysing pathogenic DNA in a blood sample. Of course, other types of biological or non-biological fluid are also analysed using an appropriate set, or combination, of reagents, test protocols, LOC variants and detection systems. Referring back to Figure 4, there are five major steps involved in analysing a biological sample, comprising sample input and preparation 288, nucleic acid extraction 290, nucleic acid incubation 291, nucleic acid amplification 292 and detection and analysis 294.
The sample input and preparation step 288 involves mixing the blood with an anticoagulant 116 and then separating pathogens from the leukocytes and erythrocytes with the pathogen dialysis section 70. As best shown in Figures 7 and 12, the blood sample enters the device via the sample inlet 68. Capillary action draws the blood sample along the cap channel 94 to the reservoir 54. Anticoagulant is released from the reservoir 54 as the sample blood flow opens its surface tension valve 118 (see Figures 15 and 22). The anticoagulant prevents the formation of clots which would block the flow.
As best shown in Figure 22, the anticoagulant 1 16 is drawn out of the reservoir 54 by capillary action and into the MST channel 90 via the downtake 92. The downtake 92
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has a capillary initiation feature (CIF) 102 to shape the geometry of the meniscus such that it does not anchor to the rim of the downtake 92. Vent holes 122 in the upper seal 82 allows air to replace the anticoagulant 1 16 as it is drawn out of the reservoir 54.
The MST channel 90 shown in Figure 22 is part of a surface tension valve 1 18. The anticoagulant 1 16 fills the surface tension valve 118 and pins a meniscus 120 to the uptake 96 to a meniscus anchor 98. Prior to use, the meniscus 120 remains pinned at the uptake 96 so the anticoagulant does not flow into the cap channel 94. When the blood flows through the cap channel 94 to the uptake 96, the meniscus 120 is removed and the anticoagulant is drawn into the flow.
Figures 15 to 21 show Inset AE which is a portion of Inset AA shown in Figure 13.
As shown in Figures 15, 16 and 17, the surface tension valve 118 has three separate MST channels 90 extending between respective downtakes 92 and uptakes 96. The number of MST channels 90 in a surface tension valve can be varied to change the flow rate of the reagent into the sample mixture. As the sample mixture and the reagents mix together by diffusion, the flow rate out of the reservoir determines the concentration of the reagent in the sample flow. Hence, the surface tension valve for each of the reservoirs is configured to match the desired reagent concentration.
The blood passes into a pathogen dialysis section 70 (see Figures 4 and 15) where target cells are concentrated from the sample using an array of apertures 164 sized according to a predetermined threshold. Cells smaller than the threshold pass through the apertures while larger cells do not pass through the apertures. Unwanted cells, which may be either the larger cells withheld by the array of apertures 164 or the smaller cells that pass through the apertures, are redirected to a waste unit 76 while the target cells continue as part of the assay.
In the pathogen dialysis section 70 described here, the pathogens from the whole blood sample are concentrated for microbial DNA analysis. The array of apertures is formed by a multitude of 3 micron diameter holes 164 fluidically connecting the input flow in the cap channel 94 to a target channel 74. The 3 micron diameter apertures 164 and the dialysis uptake holes 168 for the target channel 74 are connected by a series of dialysis MST channels 204 (best shown in Figures 15 and 21). Pathogens are small enough to pass through the 3 micron diameter apertures 164 and fill the target channel 74 via the dialysis MST channels 204. Cells larger than 3 microns, such as erythrocytes and leukocytes, stay in the waste channel 72 in the cap 46 which leads to a waste reservoir 76 (see Figure 7).
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Other aperture shapes, sizes and aspect ratios can be used to isolate specific pathogens or other target cells such as leukocytes for human DNA analysis. Greater detail on the dialysis section and dialysis variants is provided later.
Referring again to Figures 6 and 7, the flow is drawn through the target channel 74 to the surface tension valve 128 of the lysis reagent reservoir 56. The surface tension valve 128 has seven MST channels 90 extending between the lysis reagent reservoir 56 and the target channel 74. When the menisci are unpinned by the sample flow, the flow rate from all seven of the MST channels 90 will be greater than the flow rate from the anticoagulant reservoir 54 where the surface tension valve 118 has three MST channels 90 (assuming the physical characteristics of the fluids are roughly equivalent). Hence the proportion of lysis reagent in the sample mixture is greater than that of the anticoagulant.
The lysis reagent and target cells mix by diffusion in the target channel 74 within the chemical lysis section 130. A boiling-initiated valve 126 stops the flow until sufficient time has passed for diffusion and lysis to take place, releasing the genetic material from the target cells (see Figures 6 and 7). The structure and operation of the boiling-initiated valves are described in greater detail below with reference to Figures 31 and 32. Other active valve types (as opposed to passive valves such as the surface tension valve 118) have also been developed by the Applicant which may be used here instead of the boiling- initiated valve. These alternative valve designs are also described later.
When the boiling-initiated valve 126 opens, the lysed cells flow into a mixing section 131 for pre-amplification restriction digestion and linker ligation.
Referring to Figure 13, restriction enzymes, linkers and ligase are released from the reservoir 58 when the flow unpins the menisci at the surface tension valve 132 at the start of the mixing section 131. The mixture flows the length of the mixing section 131 for diffusion mixing. At the end of the mixing section 131 is downtake 134 leading into the incubator inlet channel 133 of the incubation section 114 (see Figure 13). The incubator inlet channel 133 feeds the mixture into a serpentine configuration of heated microchannels 210 which provides an incubation chamber for holding the sample during restriction digestion and ligation of the linkers (see Figures 13 and 14).
Figures 23, 24, 25, 26, 27, 28 and 29 show the layers of the LOC device 301 within
Inset AB of Figure 6. Each figure shows the sequential addition of layers forming the structures of the CMOS + MST layer 48 and the cap 46. Inset AB shows the end of the incubation section 114 and the start of the amplification section 112. As best shown in Figures 14 and 23, the flow fills the microchannels 210 of the incubation section 1 14 until
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reaching the boiling-initiated valve 106 where the flow stops while diffusion takes place. As discussed above, the microchannel 210 upstream of the boiling-initiated valve 106 becomes an incubation chamber containing the sample, restriction enzymes, ligase and linkers. The heaters 154 are then activated and held at constant temperature for a specified time for restriction digestion and linker ligation to occur.
The skilled worker will appreciate that this incubation step 291 (see Figure 4) is optional and only required for some nucleic acid amplification assay types. Furthermore, in some instances, it may be necessary to have a heating step at the end of the incubation period to spike the temperature above the incubation temperature. The temperature spike inactivates the restriction enzymes and ligase prior to entering the amplification section 1 12. Inactivation of the restriction enzymes and ligase has particular relevance when isothermal nucleic acid amplification is being employed.
Following incubation, the boiling-initiated valve 106 is activated (opened) and the flow resumes into the amplification section 112. Referring to Figures 31 and 32, the mixture fills the serpentine configuration of heated microchannels 158, which form one or more amplification chambers, until it reaches the boiling-initiated valve 108. As best shown in the schematic section view of Figure 30, amplification mix (dNTPs, primers, buffer) is released from reservoir 60 and polymerase is subsequently released from reservoir 62 into the intermediate MST channel 212 connecting the incubation and amplification sections (114 and 112 respectively).
Figures 35 to 51 show the layers of the LOC device 301 within Inset AC of Figure 6. Each figure shows the sequential addition of layers forming the structures of the CMOS + MST device 48 and the cap 46. Inset AC is at the end of the amplification section 1 12 and the start of the hybridization and detection section 52. The incubated sample, amplification mix and polymerase flow through the microchannels 158 to the boiling- initiated valve 108. After sufficient time for diffusion mixing, the heaters 154 in the microchannels 158 are activated for thermal cycling or isothermal amplification. The amplification mix goes through a predetermined number of thermal cycles or a preset amplification time to amplify sufficient target DNA. After the nucleic acid amplification process, the boiling-initiated valve 108 opens and flow resumes into the hybridization and detection section 52. The operation of boiling- initiated valves is described in more detail later.
As shown in Figure 52, the hybridization and detection section 52 has an array of hybridization chambers 1 10. Figures 52, 53, 54 and 56 show the hybridization chamber
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array 110 and individual hybridization chambers 180 in detail. At the entrance to the hybridization chamber 180 is a diffusion barrier 175 which prevents diffusion of the target nucleic acid, probe strands and hybridized probes between the hybridization chambers 180 during hybridization so as to prevent erroneous hybridization detection results. The diffusion barriers 175 present a flow-path-length that is long enough to prevent the target sequences and probes diffusing out of one chamber and contaminating another chamber within the time taken for the probes and nucleic acids to hybridize and the signal to be detected, thus avoiding an erroneous result.
Another mechanism to prevent erroneous readings is to have identical probes in a number of the hybridization chambers. The CMOS circuitry 86 derives a single result from the photodiodes 184 corresponding to the hybridization chambers 180 that contain identical probes. Anomalous results can be disregarded or weighted differently in the derivation of the single result.
The thermal energy required for hybridization is provided by CMOS-controlled heaters 182 (described in more detail below). After the heater is activated, hybridization occurs between complementary target-probe sequences. The LED driver 29 in the CMOS circuitry 86 signals the LED 26 located in the test module 10 to illuminate. These probes only fluoresce when hybridization has occurred thereby avoiding washing and drying steps that are typically required to remove unbound strands. Hybridization forces the stem-and- loop structure of the FRET probes 186 to open, which allows the fluorophore to emit fluorescent energy in response to the LED excitation light, as discussed in greater detail later. Fluorescence is detected by a photodiode 184 in the CMOS circuitry 86 underlying each hybridization chamber 180 (see hybridization chamber description below). The photodiodes 184 for all hybridization chambers and associated electronics collectively form the photosensor 44 (see Figure 68). In other embodiments, the photosensor may be an array of charge coupled devices (CCD array). The detected signal from the photodiodes 184 is amplified and converted to a digital output which is analyzed by the test module reader 12. Further details of the detection method are described later.
ADDITIONAL DETAILS FOR THE LOC DEVICE MODULARITY OF THE DESIGN
The LOC device 301 has many functional sections, including the reagent reservoirs 54, 56, 58, 60 and 62, the dialysis section 70, lysis section 130, incubation section 1 14, and
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amplification section 112, valve types, the humidifier and humidity sensor. In other embodiments of the LOC device, these functional sections can be omitted, additional functional sections can be added or the functional sections can be used for alternative purposes to those described above.
For example, the incubation section 1 14 can be used as the first amplification section 112 of a tandem amplification assay system, with the chemical lysis reagent reservoir 56 being used to add the first amplification mix of primers, dNTPs and buffer and reagent reservoir 58 being used for adding the reverse transcriptase and/or polymerase. A chemical lysis reagent can also be added to the reservoir 56 along with the amplification mix if chemical lysis of the sample is desired or, alternatively, thermal lysis can occur in the incubation section by heating the sample for a predetermined time. In some embodiments, an additional reservoir can be incorporated immediately upstream of reservoir 58 for the mix of primers, dNTPs and buffer if there is a requirement for chemical lysis and a separation of this mix from the chemical lysis reagent is desired.
In some circumstances it may be desirable to omit a step, such as the incubation step 291. In this case, a LOC device can be specifically fabricated to omit the reagent reservoir 58 and incubation section 1 14, or the reservoir can simply not be loaded with reagents or the active valves, if present, not activated to dispense the reagents into the sample flow, and the incubation section then simply becomes a channel to transport the sample from the lysis section 130 to the amplification section 112. The heaters are independently operable and therefore, where reactions are dependent on heat, such as thermal lysis, programming the heaters not to activate during this step ensures thermal lysis does not occur in LOC devices that do not require it. The dialysis section 70 can be located at the beginning of the fluidic system within the micro fluidic device as shown in Figure 4 or can be located anywhere else within the microfluidic device. For example, dialysis after the amplification phase 292 to remove cellular debris prior to the
hybridization and detection step 294 may be beneficial in some circumstances.
Alternatively, two or more dialysis sections can be incorporated at any location throughout the LOC device. Similarly, it is possible to incorporate additional amplification sections 1 12 to enable multiple targets to be amplified in parallel or in series prior to being detected in the hybridization chamber arrays 110 with specific nucleic acid probes. For analysis of samples like whole blood, in which dialysis is not required, the dialysis section 70 is simply omitted from the sample input and preparation section 288 of the LOC design. In some cases, it is not necessary to omit the dialysis section 70 from the LOC device even if
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the analysis does not require dialysis. If there is no geometric hindrance to the assay by the existence of a dialysis section, a LOC with the dialysis section 70 in the sample input and preparation section can still be used without a loss of the required functionality.
Furthermore, the detection section 294 may encompass proteomic chamber arrays which are identical to the hybridization chamber arrays but are loaded with probes designed to conjugate or hybridize with sample target proteins present in non-amplified sample instead of nucleic acid probes designed to hybridize to target nucleic acid sequences.
It will be appreciated that the LOC devices fabricated for use in this diagnostic system are different combinations of functional sections selected in accordance with the particular LOC application. The vast majority of functional sections are common to many of the LOC devices and the design of additional LOC devices for new application is a matter of compiling an appropriate combination of functional sections from the extensive selection of functional sections used in the existing LOC devices.
Only a small number of the LOC devices are shown in this description and some more are shown schematically to illustrate the design flexibility of the LOC devices fabricated for this system. The person skilled in the art will readily recognise that the LOC devices shown in this description are not an exhaustive list and many additional LOC designs are a matter of compiling the appropriate combination of functional sections. SAMPLE TYPES
LOC variants can accept and analyze the nucleic acid or protein content of a variety of sample types in liquid form including, but not limited to, blood and blood products, saliva, cerebrospinal fluid, urine, semen, amniotic fluid, umbilical cord blood, breast milk, sweat, pleural effusion, tear, pericardial fluid, peritoneal fluid, environmental water samples and drink samples. Amplicon obtained from macroscopic nucleic acid amplification can also be analysed using the LOC device; in this case, all the reagent reservoirs will be empty or configured not to release their contents, and the dialysis, lysis, incubation and amplification sections will be used solely to transport the sample from the sample inlet 68 to the hybridization chambers 180 for nucleic acid detection, as described above.
For some sample types, a pre-processing step is required, for example semen may need to be liquefied and mucus may need to be pre-treated with an enzyme to reduce the viscosity prior to input into the LOC device.
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SAMPLE INPUT
Referring to Figures 1 and 12, the sample is added to the macroreceptacle 24 of the test module 10. The macroreceptacle 24 is a truncated cone which feeds into the inlet 68 of the LOC device 301 by capillary action. Here it flows into the 64 μηι wide x 60 μιη deep cap channel 94 where it is drawn towards the anticoagulant reservoir 54, also by capillary action.
REAGENT RESERVOIRS
The small volumes of reagents required by the assay systems using microfluidic devices, such as LOC device 301, permit the reagent reservoirs to contain all reagents necessary for the biochemical processing with each of the reagent reservoirs having a small volume. This volume is easily less than 1,000,000,000 cubic microns, in the vast majority of cases less than 300,000,000 cubic microns, typically less than 70,000,000 cubic microns and in the case of the LOC device 301 shown in the drawings, less than 20,000,000 cubic microns. DIALYSIS SECTION
Referring to Figures 15 to 21, 33 and 34, the pathogen dialysis section 70 is designed to concentrate pathogenic target cells from the sample. As previously described, a plurality of apertures in the form of 3 micron diameter holes 164 in the roof layer 66 filter the target cells from the bulk of the sample. As the sample flows past the 3 micron diameter apertures 164, microbial pathogens pass through the holes into a series of dialysis MST channels 204 and flow back up into the target channel 74 via 16μιη dialysis uptake holes 168 (see Figures 33 and 34). The remainder of the sample (erythrocytes and so on) stay in the cap channel 94. Downstream of the pathogen dialysis section 70, the cap channel 94 becomes the waste channel 72 leading to the waste reservoir 76. For biological samples of the type that generate a substantial amount of waste, a foam insert or other porous element 49 within the outer casing 13 of the test module 10 is configured to be in fluid communication with the waste reservoir 76 (see Figure 1).
The pathogen dialysis section 70 functions entirely on capillary action of the fluid sample. The 3 micron diameter apertures 164 at the upstream end of the pathogen dialysis section 70 have capillary initiation features (CIFs) 166 (see Figure 33) so that the fluid is drawn down into the dialysis MST channel 204 beneath. The first uptake hole 198 for the
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target channel 74 also has a CIF 202 (see Figure 15) to avoid the flow simply pinning a meniscus across the dialysis uptake holes 168.
The small constituents dialysis section 682 schematically shown in Figure 81 can have a similar structure to the pathogen dialysis section 70. The small constituents dialysis section separates any small target cells or molecules from a sample by sizing (and, if necessary, shaping) apertures suitable for allowing the small target cells or molecules to pass into the target channel and continue for further analysis. Larger sized cells or molecules are removed to a waste reservoir 766. Thus, the LOC device 30 (see Figures 1 and 104) is not limited to separating pathogens that are less than 3 μπι in size, but can be used to separate cells or molecules of any size desired.
LYSIS SECTION
Referring back to Figures 7, 1 1 and 13, the genetic material in the sample is released from the cells by a chemical lysis process. As described above, a lysis reagent from the lysis reservoir 56 mixes with the sample flow in the target channel 74 downstream of the surface tension valve 128 for the lysis reservoir 56. However, some diagnostic assays are better suited to a thermal lysis process, or even a combination of chemical and thermal lysis of the target cells. The LOC device 301 accommodates this with the heated microchannels 210 of the incubation section 1 14. The sample flow fills the incubation section 114 and stops at the boiling-initiated valve 106. The incubation microchannels 210 heat the sample to a temperature at which the cellular membranes are disrupted.
In some thermal lysis applications, an enzymatic reaction in the chemical lysis section 130 is not necessary and the thermal lysis completely replaces the enzymatic reaction in the chemical lysis section 130. BOILING-INITIATED VALVE
As discussed above, the LOC device 301 has three boiling-initiated valves 126, 106 and 108. The location of these valves is shown in Figure 6. Figure 31 is an enlarged plan view of the boiling-initiated valve 108 in isolation at the end of the heated microchannels 158 of the amplification section 1 12.
The sample flow 1 19 is drawn along the heated microchannels 158 by capillary action until it reaches the boiling-initiated valve 108. The leading meniscus 120 of the
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sample flow pins at a meniscus anchor 98 at the valve inlet 146. The geometry of the meniscus anchor 98 stops the advancing meniscus to arrest the capillary flow. As shown in Figures 31 and 32, the meniscus anchor 98 is an aperture provided by an uptake opening from the MST channel 90 to the cap channel 94. Surface tension in the meniscus 120 keeps the valve closed. An annular heater 152 is at the periphery of the valve inlet 146. The annular heater 152 is CMOS-controlled via the boiling- initiated valve heater contacts 153.
To open the valve, the CMOS circuitry 86 sends an electrical pulse to the valve heater contacts 153. The annular heater 152 resistively heats until the liquid sample 1 19 boils. The boiling unpins the meniscus 120 from the valve inlet 146 and initiates wetting of the cap channel 94. Once wetting the cap channel 94 begins, capillary flow resumes. The fluid sample 119 fills the cap channel 94 and flows through the valve downtake 150 to the valve outlet 148 where capillary driven flow continues along the amplification section exit channel 160 into the hybridization and detection section 52. Liquid sensors 174 are placed before and after the valve for diagnostics.
It will be appreciated that once the boiling-initiated valves are opened, they cannot be re-closed. However, as the LOC device 301 and the test module 10 are single-use devices, re-closing the valves is unnecessary.
INCUBATION SECTION AND NUCLEIC ACID AMPLIFICATION SECTION Figures 6, 7, 13, 14, 23, 24, 25, 35 to 45, 50 and 51 show the incubation section 114 and the amplification section 112. The incubation section 114 has a single, heated incubation microchannel 210 etched in a serpentine pattern in the MST channel layer 100 from the downtake opening 134 to the boiling-initiated valve 106 (see Figures 13 and 14). Control over the temperature of the incubation section 114 enables enzymatic reactions to take place with greater efficiency. Similarly, the amplification section 112 has a heated amplification microchannel 158 in a serpentine configuration leading from the boiling- initiated valve 106 to the boiling-initiated valve 108 (see Figures 6 and 14). These valves arrest the flow to retain the target cells in the heated incubation or amplification microchannels 210 or 158 while mixing, incubation and nucleic acid amplification takes place. The serpentine pattern of the microchannels also facilitates (to some extent) mixing of the target cells with reagents.
In the incubation section 114 and the amplification section 1 12, the sample cells and the reagents are heated by the heaters 154 controlled by the CMOS circuitry 86 using
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pulse width modulation (PWM). Each meander of the serpentine configuration of the heated incubation microchannel 210 and amplification microchannel 158 has three separately operable heaters 154 extending between their respective heater contacts 156 (see Figure 14) which provides for the two-dimensional control of input heat flux density. As best shown in Figure 51 , the heaters 154 are supported on the roof layer 66 and embedded in the lower seal 64. The heater material is TiAl but many other conductive metals would be suitable. The elongate heaters 154 are parallel with the longitudinal extent of each channel section that forms the wide meanders of the serpentine shape. In the amplification section 112, each of the wide meanders can operate as separate PC chambers via individual heater control.
The small volumes of amplicon required by the assay systems using microfluidic devices, such as LOC device 301, permit low amplification mixture volumes for amplification in amplification section 112. This volume is easily less than 400 nanoliters, in the vast majority of cases less than 170 nanoliters, typically less than 70 nanoliters and in the case of the LOC device 301, between 2 nanoliters and 30 nanoliters.
INCREASED RATES OF HEATING AND GREATER DIFFUSIVE MIXING
The small cross section of each channel section increases the heating rate of the amplification fluid mix. All the fluid is kept a relatively short distance from the heater 154. Reducing the channel cross section (that is the amplification microchannel 158 cross section) to less than 100,000 square microns achieves appreciably higher heating rates than that provided by more 'macro-scale' equipment. Lithographic fabrication techniques allow the amplification microchannel 158 to have a cross sectional area transverse to the flow- path less than 16,000 square microns which gives substantially higher heating rates.
Feature sizes on the order of 1 micron are readily achievable with lithographic techniques. If very little amplicon is needed (as is the case in the LOC device 301), the cross sectional area can be reduced to less than 2,500 square microns. For diagnostic assays with 1,000 to 2,000 probes on the LOC device, and a requirement of 'sample-in, answer out' in less than 1 minute, a cross sectional area transverse to the flow of between 400 square microns and 1 square micron is adequate.
The heater element in the amplification microchannel 158 heats the nucleic acid sequences at a rate more than 80 Kelvin (K) per second, in the vast majority of cases at a rate greater than 100 K per second. Typically, the heater element heats the nucleic acid sequences at a rate more than 1,000 K per second and in many cases, the heater element
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heats the nucleic acid sequences at a rate more than 10,000 per second. Commonly, based on the demands of the assay system, the heater element heats the nucleic acid sequences at a rate more than 100,000 K per second, more than 1,000,000 K per second more than 10,000,000 K per second, more than 20,000,000 per second, more than 40,000,000 K per second, more than 80,000,000 K per second and more than 160,000,000 K per second.
A small cross-sectional area channel is also beneficial for diffusive mixing of any reagents with the sample fluid. Before diffusive mixing is complete, diffusion of one liquid into the other is greatest near the interface between the two. Concentration decreases with distance from the interface. Using microchannels with relatively small cross sections transverse to the flow direction, keeps both fluid flows close to the interface for more rapid diffusive mixing. Reducing the channel cross section to less than 100,000 square microns achieves appreciably higher mixing rates than that provided by more 'macro-scale' equipment. Lithographic fabrication techniques allows microchannels with a cross sectional area transverse to the flow-path less than 16000 square microns which gives significantly higher mixing rates. If small volumes are needed (as is the case in the LOC device 301), the cross sectional area can be reduced to less than 2500 square microns. For diagnostic assays with 1000 to 2000 probes on the LOC device, and a requirement of 'sample-in, answer out' in less than 1 minute, a cross sectional area transverse to the flow of between 400 square microns and 1 square micron is adequate.
SHORT THERMAL CYCLE TIMES
Keeping the sample mixture proximate to the heaters, and using very small fluid volumes allows rapid thermal cycling during the nucleic acid amplification process. Each thermal cycle (i.e. denaturing, annealing and primer extension) is completed in less than 30 seconds for target sequences up to 150 base pairs (bp) long. In the vast majority of diagnostic assays, the individual thermal cycle times are less than 11 seconds, and a large proportion are less than 4 seconds. LOC devices 30 with some of the most common diagnostic assays have thermal cycles time between 0.45 seconds to 1.5 seconds for target sequences up to 150 bp long. Thermal cycling at this rate allows the test module to complete the nucleic acid amplification process in much less than 10 minutes; often less than 220 seconds. For most assays, the amplification section generates sufficient amplicon in less than 80 seconds from the sample fluid entering the sample inlet. For a great many assays, sufficient amplicon is generated in 30 seconds.
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Upon completion of a preset number of amplification cycles, the amplicon is fed into the hybridization and detection section 52 via the boiling-initiated valve 108.
HYBRIDIZATION CHAMBERS
Figures 52, 53, 54, 56 and 57 show the hybridization chambers 180 in the hybridization chamber array 1 10. The hybridization and detection section 52 has a 24 x 45 array 110 of hybridization chambers 180, each with hybridization-responsive FRET probes 186, heater element 182 and an integrated photodiode 184. The photodiode 184 is incorporated for detection of fluorescence resulting from the hybridization of a target nucleic acid sequence or protein with the FRET probes 186. Each photodiode 184 is independently controlled by the CMOS circuitry 86. Any material between the FRET probes 186 and the photodiode 184 must be transparent to the emitted light. Accordingly, the wall section 97 between the probes 186 and the photodiode 184 is also optically transparent to the emitted light. In the LOC device 301, the wall section 97 is a thin (approximately 0.5 micron) layer of silicon dioxide.
Incorporation of a photodiode 184 directly beneath each hybridization chamber 180 allows the volume of probe-target hybrids to be very small while still generating a detectable fluorescence signal (see Figure 54). The small amounts permit small volume hybridization chambers. A detectable amount of probe-target hybrid requires a quantity of probe, prior to hybridization, which is easily less than 270 picograms (corresponding to 900,000 cubic microns), in the vast majority of cases less than 60 picograms
(corresponding to 200,000 cubic microns), typically less than 12 picograms (corresponding to 40,000 cubic microns) and in the case of the LOC device 301 shown in the
accompanying figures, less than 2.7 picograms (corresponding to a chamber volume of 9,000 cubic microns). Of course, reducing the size of the hybridization chambers allows a higher density of chambers and therefore more probes on the LOC device. In LOC device 301, the hybridization section has more than 1,000 chambers in an area of 1,500 microns by 1,500 microns (i.e. less than 2,250 square microns per chamber). Smaller volumes also reduce the reaction times so that hybridization and detection is faster. An additional advantage of the small amount of probe required in each chamber is that only very small quantities of probe solution need to be spotted into each chamber during production of the LOC device. Embodiments of the LOC device according to the invention can be spotted using a probe solution volume of 1 picoliter or less.
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After nucleic acid amplification, boiling-initiated valve 108 is activated and the amplicon flows along the flow-path 176 and into each of the hybridization chambers 180 (see Figures 52 and 56). An end-point liquid sensor 178 indicates when the hybridization chambers 180 are filled with amplicon and the heaters 182 can be activated.
After sufficient hybridization time, the LED 26 (see Figure 2) is activated. The opening in each of the hybridization chambers 180 provides an optical window 136 for exposing the FRET probes 186 to the excitation radiation (see Figures 52, 54 and 56). The LED 26 is illuminated for a sufficiently long time in order to induce a fluorescence signal from the probes with high intensity. During excitation, the photodiode 184 is shorted. After a pre-programmed delay 300 (see Figure 2), the photodiode 184 is enabled and fluorescence emission is detected in the absence of the excitation light. The incident light on the active area 185 of the photodiode 184 (see Figure 54) is converted into a photocurrent which can then be measured using CMOS circuitry 86.
The hybridization chambers 180 are each loaded with probes for detecting a single target nucleic acid sequence. Each hybridization chambers 180 can be loaded with probes to detect over 1,000 different targets if desired. Alternatively, many or all the
hybridization chambers can be loaded with the same probes to detect the same target nucleic acid repeatedly. Replicating the probes in this way throughout the hybridization chamber array 1 10 leads to increased confidence in the results obtained and the results can be combined by the photodiodes adjacent those hybridization chambers to provide a single result if desired. The person skilled in the art will recognise that it is possible to have from one to over 1,000 different probes on the hybridization chamber array 1 10, depending on the assay specification.
HUMIDIFIER AND HUMIDITY SENSOR Inset AG of Figure 6 indicates the position of the humidifier 196. The humidifier prevents evaporation of the reagents and probes during operation of the LOC device 301. As best shown in the enlarged view of Figure 55, a water reservoir 188 is fluidically connected to three evaporators 190. The water reservoir 188 is filled with molecular biology-grade water and sealed during manufacturing. As best shown in Figures 55 and 74, water is drawn into three downtakes 194 and along respective water supply channels 192 by capillary action to a set of three uptakes 193 at the evaporators 190. A meniscus pins at each uptake 193 to retain the water. The evaporators have annular shaped heaters 191 which encircle the uptakes 193. The annular heaters 191 are connected to the CMOS
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circuitry 86 by the conductive columns 376 to the top metal layer 195 (see Figure 37). Upon activation, the annular heaters 191 heat the water causing evaporation and humidifying the device surrounds.
The position of the humidity sensor 232 is also shown in Figure 6. However, as best shown in the enlarged view of Inset AH in Figure 67, the humidity sensor has a capacitive comb structure. A lithographically etched first electrode 296 and a
lithographically etched second electrode 298 face each other such that their teeth are interleaved. The opposed electrodes form a capacitor with a capacitance that can be monitored by the CMOS circuitry 86. As the humidity increases, the permittivity of the air gap between the electrodes increases, so that the capacitance also increases. The humidity sensor 232 is adjacent the hybridization chamber array 110 where humidity measurement is most important to slow evaporation from the solution containing the exposed probes.
FEEDBACK SENSORS
Temperature and liquid sensors are incorporated throughout the LOC device 301 to provide feedback and diagnostics during device operation. Referring to Figure 35, nine temperature sensors 170 are distributed throughout the amplification section 1 12.
Likewise, the incubation section 114 also has nine temperature sensors 170. These sensors each use a 2x2 array of bipolar junction transistors (BJTs) to monitor the fluid temperature and provide feedback to the CMOS circuitry 86. The CMOS circuitry 86 uses this to precisely control the thermal cycling during the nucleic acid amplification process and any heating during thermal lysis and incubation.
In the hybridization chambers 180, the CMOS circuitry 86 uses the hybridization heaters 182 as temperature sensors (see Figure 56). The electrical resistance of the hybridization heaters 182 is temperature dependent and the CMOS circuitry 86 uses this to derive a temperature reading for each of the hybridization chambers 180.
The LOC device 301 also has a number of MST channel liquid sensors 174 and cap channel liquid sensors 208. Figure 35 shows a line of MST channel liquid sensors 174 at one end of every other meander in the heated microchannel 158. As best shown in Figure 37, the MST channel liquid sensors 174 are a pair of electrodes formed by exposed areas of the top metal layer 195 in the CMOS structure 86. Liquid closes the circuit between the electrodes to indicate its presence at the sensor's location.
Figure 25 shows an enlarged perspective of cap channel liquid sensors 208.
Opposing pairs of TiAl electrodes 218 and 220 are deposited on the roof layer 66.
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Between the electrodes 218 and 220 is a gap 222 to hold the circuit open in the absence of liquid. The presence of liquid closes the circuit and the CMOS circuitry 86 uses this feedback to monitor the flow.
GRAVITATIONAL INDEPENDENCE The test modules 10 are orientation independent. They do not need to be secured to a flat stable surface in order to operate. Capillary driven fluid flows and a lack of external plumbing into ancillary equipment allow the modules to be truly portable and simply plugged into a similarly portable hand held reader such as a mobile telephone. Having a gravitationally independent operation means the test modules are also accelerationally independent to all practical extents. They are resistant to shock and vibration and will operate on moving vehicles or while the mobile telephone is being carried around.
NUCLEIC ACID AMPLIFICATION VARIANTS
DIRECT PCR
Traditionally, PCR requires extensive purification of the target DNA prior to preparation of the reaction mixture. However, with appropriate changes to the chemistry and sample concentration, it is possible to perform nucleic acid amplification with minimal DNA purification, or direct amplification. When the nucleic acid amplification process is PCR, this approach is called direct PCR. In LOC devices where nucleic acid amplification is performed at a controlled, constant temperature, the approach is direct isothermal amplification. Direct nucleic acid amplification techniques have considerable advantages for use in LOC devices, particularly relating to simplification of the required fluidic design. Adjustments to the amplification chemistry for direct PCR or direct isothermal amplification include increased buffer strength, the use of polymerases which have high activity and processivity, and additives which chelate with potential polymerase inhibitors. Dilution of inhibitors present in the sample is also important.
To take advantage of direct nucleic acid amplification techniques, the LOC device designs incorporate two additional features. The first feature is reagent reservoirs (for example reservoir 58 in Figure 8) which are appropriately dimensioned to supply a sufficient quantity of amplification reaction mix, or diluent, so that the final concentrations of sample components which might interfere with amplification chemistry are low enough to permit successful nucleic acid amplification. The desired dilution of non-cellular
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sample components is in the range of 5X to 20X. Different LOC structures, for example the pathogen dialysis section 70 in Figure 4, are used when appropriate to ensure that the concentration of target nucleic acid sequences is maintained at a high enough level for amplification and detection. In this embodiment, further illustrated in Figure 6, a dialysis section which effectively concentrates pathogens small enough to be passed into the amplification section 292 is employed upstream of the sample extraction section 290, and rejects larger cells to a waste receptacle 76. In another embodiment, a dialysis section is used to selectively deplete proteins and salts in blood plasma while retaining cells of interest.
The second LOC structural feature which supports direct nucleic acid amplification is design of channel aspect ratios to adjust the mixing ratio between the sample and the amplification mix components. For example, to ensure dilution of inhibitors associated with the sample in the preferred 5X - 20X range through a single mixing step, the length and cross-section of the sample and reagent channels are designed such that the sample channel, upstream of the location where mixing is initiated, constitutes a flow impedance 4X - 19X higher than the flow impedance of the channels through which the reagent mixture flows. Control over flow impedances in microchannels is readily achieved through control over the design geometry. The flow impedance of a microchannel increases linearly with the channel length, for a constant cross-section. Importantly for mixing designs, flow impedance in microchannels depends more strongly on the smallest cross-sectional dimension. For example, the flow impedance of a microchannel with rectangular cross-section is inversely proportional to the cube of the smallest perpendicular dimension, when the aspect ratio is far from unity.
REVERSE-TRANSCRIPTASE PCR (RT-PCR) Where the sample nucleic acid species being analysed or extracted is RNA, such as from RNA viruses or messenger RNA, it is first necessary to reverse transcribe the RNA into complementary DNA (cDNA) prior to PCR amplification. The reverse transcription reaction can be performed in the same chamber as the PCR (one-step RT-PCR) or it can be performed as a separate, initial reaction (two-step RT-PCR). In the LOC variants described herein, a one-step RT-PCR can be performed simply by adding the reverse transcriptase to reagent reservoir 62 along with the polymerase and programming the heaters 1 4 to cycle firstly for the reverse transcription step and then progress onto the nucleic acid amplification step. A two-step RT-PCR could also be easily achieved by
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utilizing the reagent reservoir 58 to store and dispense the buffers, primers, dNTPs and reverse transcriptase and the incubation section 1 14 for the reverse transcription step followed by amplification in the normal way in the amplification section 112.
ISOTHERMAL NUCLEIC ACID AMPLIFICATION For some applications, isothermal nucleic acid amplification is the preferred method of nucleic acid amplification, thus avoiding the need to repetitively cycle the reaction components through various temperature cycles but instead maintaining the amplification section at a constant temperature, typically around 37°C to 41°C. A number of isothermal nucleic acid amplification methods have been described, including Strand Displacement Amplification (SDA), Transcription Mediated Amplification (TMA), Nucleic Acid Sequence Based Amplification (NASBA), Recombinase Polymerase Amplification (RPA), Helicase-Dependent isothermal DNA Amplification (HDA), Rolling Circle Amplification (RCA), Ramification Amplification (RAM) and Loop-mediated Isothermal Amplification (LAMP), and any of these, or other isothermal amplification methods, can be employed in particular embodiments of the LOC device described herein.
In order to perform isothermal nucleic acid amplification, the reagent reservoirs 60 and 62 adjoining the amplification section will be loaded with the appropriate reagents for the specified isothermal method instead of PCR amplification mix and polymerase. For example, for SDA, reagent reservoir 60 contains amplification buffer, primers and dNTPs and reagent reservoir 62 contains an appropriate nickase enzyme and Exo- DNA polymerase. For RPA, reagent reservoir 60 contains the amplification buffer, primers, dNTPs and recombinase proteins, with reagent reservoir 62 containing a strand displacing DNA polymerase such as Bsu. Similarly, for HDA, reagent reservoir 60 contains amplification buffer, primers and dNTPs and reagent reservoir 62 contains an appropriate DNA polymerase and a helicase enzyme to unwind the double stranded DNA strand instead of using heat. The skilled person will appreciate that the necessary reagents can be split between the two reagent reservoirs in any manner appropriate for the nucleic acid amplification process.
For amplification of viral nucleic acids from RNA viruses such as HIV or hepatitis C virus, NASBA or TMA is appropriate as it is unnecessary to first transcribe the RNA to cDNA. In this example, reagent reservoir 60 is filled with amplification buffer, primers and dNTPs and reagent reservoir 62 is filled with RNA polymerase, reverse transcriptase and, optionally, RNase H.
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For some forms of isothermal nucleic acid amplification it may be necessary to have an initial denaturation cycle to separate the double stranded DNA template, prior to maintaining the temperature for the isothermal nucleic acid amplification to proceed. This is readily achievable in all embodiments of the LOC device described herein, as the temperature of the mix in the amplification section 112 can be carefully controlled by the heaters 154 in the amplification microchannels 158 (see Figure 14).
Isothermal nucleic acid amplification is more tolerant of potential inhibitors in the sample and, as such, is generally suitable for use where direct nucleic acid amplification from the sample is desired. Therefore, isothermal nucleic acid amplification is sometimes useful in LOC variant XLIII 673, LOC variant XLIV 674 and LOC variant XLVII 677, amongst others, shown in Figures 82, 83 and 84, respectively. Direct isothermal amplification may also be combined with one or more pre-amplification dialysis steps 70, 686 or 682 as shown in Figures 82 and 84 and/or a pre-hybridization dialysis step 682 as indicated in Figure 83 to help partially concentrate the target cells in the sample before nucleic acid amplification or remove unwanted cellular debris prior to the sample entering the hybridization chamber array 110, respectively. The person skilled in the art will appreciate that any combination of pre-amplification dialysis and pre-hybridization dialysis can be used.
Isothermal nucleic acid amplification can also be performed in parallel
amplification sections such as those schematically represented in Figures 78, 79 and 80, multiplexed and some methods of isothermal nucleic acid amplification, such as LAMP, are compatible with an initial reverse transcription step to amplify RNA.
ADDITIONAL DETAILS ON THE FLUORESCENCE DETECTION SYSTEM
Figures 58 and 59 show the hybridization-responsive FRET probes 236. These are often referred to as molecular beacons and are stem-and-loop probes, generated from a single strand of nucleic acid, that fluoresce upon hybridization to complementary nucleic acids. Figure 58 shows a single FRET probe 236 prior to hybridization with a target nucleic acid sequence 238. The probe has a loop 240, stem 242, a fluorophore 246 at the 5' end, and a quencher 248 at the 3' end. The loop 240 consists of a sequence complementary to the target nucleic acid sequence 238. Complementary sequences on either side of the probe sequence anneal together to form the stem 242.
In the absence of a complementary target sequence, the probe remains closed as shown in Figure 58. The stem 242 keeps the fluorophore-quencher pair in close proximity
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to each other, such that significant resonant energy transfer can occur between them, substantially eliminating the ability of the fluorophore to fluoresce when illuminated with the excitation light 244.
Figure 59 shows the FRET probe 236 in an open or hybridized configuration. Upon hybridization to a complementary target nucleic acid sequence 238, the stem-and- loop structure is disrupted, the fluorophore and quencher are spatially separated, thus restoring the ability of the fluorophore 246 to fluoresce. The fluorescence emission 250 is optically detected as an indication that the probe has hybridized.
The probes hybridize with very high specificity with complementary targets, since the stem helix of the probe is designed to be more stable than a probe-target helix with a single nucleotide that is not complementary. Since double-stranded DNA is relatively rigid, it is sterically impossible for the probe-target helix and the stem helix to coexist.
PRIMER-LINKED PROBES
Primer-linked, stem-and-loop probes and primer-linked, linear probes, otherwise known as scorpion probes, are an alternative to molecular beacons and can be used for real-time and quantitative nucleic acid amplification in the LOC device. Real-time amplification could be performed directly in the hybridization chambers of the LOC device. The benefit of using primer-linked probes is that the probe element is physically linked to the primer, thus only requiring a single hybridization event to occur during the nucleic acid amplification rather than separate hybridizations of the primers and probes being required. This ensures that the reaction is effectively instantaneous and results in stronger signals, shorter reaction times and better discrimination than when using separate primers and probes. The probes (along with polymerase and the amplification mix) would be deposited into the hybridization chambers 180 during fabrication and there would be no need for a separate amplification section on the LOC device. Alternatively, the amplification section is left unused or used for other reactions.
PRIMER-LINKED LINEAR PROBE
Figures 85 and 86 show a primer-linked linear probe 692 during the initial round of nucleic acid amplification and in its hybridized configuration during subsequent rounds of nucleic acid amplification, respectively. Referring to Figure 85, the primer-linked linear probe 692 has a double-stranded stem segment 242. One of the strands incorporates the primer linked probe sequence 696 which is homologous to a region on the target nucleic
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acid 696 and is labelled on its 5' end with fluorophore 246, and linked on its 3' end to an oligonucleotide primer 700 via an amplification blocker 694. The other strand of the stem 242 is labelled at its 3□ end with a quencher moiety 248. After an initial round of nucleic acid amplification has completed, the probe can loop around and hybridize to the extended strand with the, now complementary, sequence 698. During the initial round of nucleic acid amplification, the oligonucleotide primer 700 anneals to the target DNA 238 (Figure 85) and is then extended, forming a DNA strand containing both the probe sequence and the amplification product. The amplification blocker 694 prevents the polymerase from reading through and copying the probe region 696. Upon subsequent denaturation, the extended oligonucleotide primer 700/template hybrid is dissociated and so is the double stranded stem 242 of the primer-linked linear probe, thus releasing the quencher 248. Once the temperature decreases for the annealing and extension steps, the primer linked probe sequence 696 of the primer-linked linear probe curls around and hybridizes to the amplified complementary sequence 698 on the extended strand and fluorescence is detected indicating the presence of the target DNA. Non-extended primer- linked linear probes retain their double-stranded stem and fluorescence remains quenched. This detection method is particularly well suited for fast detection systems as it relies on a single-molecule process.
PRIMER-LINKED STEM-AND-LOOP PROBES Figures 87A to 87F show the operation of a primer-linked stem-and-loop probe
704. Referring to Figure 87A, the primer-linked stem-and-loop probe 704 has a stem 242 of complementary double-stranded DNA and a loop 240 which incorporates the probe sequence. One of the stem strands 708 is labelled at its 5' end with fluorophore 246. The other strand 710 is labelled with a 3 '-end quencher 248 and carries both the amplification blocker 694 and oligonucleotide primer 700. During the initial denaturation phase (see
Figure 87B), the strands of the target nucleic acid 238 separate, as does the stem 242 of the primer-linked, stem-and-loop probe 704. When the temperature cools for the annealing phase (see Figure 87C), the oligonucleotide primer 700 on the primer-linked stem-and-loop probe 704 hybridizes to the target nucleic acid sequence 238. During extension (see Figure 87D) the complement 706 to the target nucleic acid sequence 238 is synthesized forming a DNA strand containing both the probe sequence 704 and the amplified product. The amplification blocker 694 prevents the polymerase from reading through and copying the probe region 704. When the probe next anneals, following denaturation, the probe
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sequence of the loop segment 240 of the primer-linked stem-and-loop probe (see Figure 87F) anneals to the complementary sequence 706 on the extended strand. This configuration leaves the fluorophore 246 relatively remote from the quencher 248, resulting in a significant increase in fluorescence emission. CONTROL PROBES
The hybridization chamber array 110 includes some hybridization chambers 180 with positive and negative control probes used for assay quality control. Figures 100 and
101 schematically illustrate negative control probes without a fluorophore 796, and Figures
102 and 103 are sketches of positive control probes without a quencher 798. The positive and negative control probes have a stem-and-loop structure like the FRET probes described above. However, a fluorescence signal 250 will always be emitted from positive control probes 798 and no fluorescence signal 250 is ever emitted from negative control probes 796, regardless of whether the probes hybridize into an open configuration or remain closed.
Referring to Figures 100 and 101, the negative control probe 796 has no fluorophore (and may or may not have a quencher 248). Hence, whether the target nucleic acid sequence 238 hybridizes with the probe (see Figure 101), or the probe remains in its stem-and-loop configuration (see Figure 100), the response to the excitation light 244 is negligible. Alternatively, the negative control probe 796 could be designed so that it always remains quenched. For example, by synthesizing the loop 240 to have a probe sequence that will not hybridize to any nucleic acid sequence within the sample under investigation, the stem 242 of the probe molecule will re-hybridize to itself and the fluorophore and quencher will remain in close proximity and no appreciable fluorescence signal will be emitted. This negative control signal would correspond to low level emissions from hybridization chambers 180 in which the probes has not hybridized but the quencher does not quench all emissions from the reporter.
Conversely, the positive control probe 798 is constructed without a quencher as illustrated in Figures 102 and 103. Nothing quenches the fluorescence emission 250 from the fluorophore 246 in response to the excitation light 244 regardless of whether the positive control probe 798 hybridizes with the target nucleic acid sequence 238.
Figure 52 shows a possible distribution of the positive and negative control probes (378 and 380 respectively) throughout the hybridization chamber array 110. The control probes 378 and 380 are placed in hybridization chambers 180 positioned in a line across
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the hybridization chamber array 110. However, the arrangement of the control probes within the array is arbitrary (as is the configuration of the hybridization chamber array 110).
FLUO OPHORE DESIGN Fluorophores with long fluorescence lifetimes are required in order to allow enough time for the excitation light to decay to an intensity below that of the fluorescence emission at which time the photosensor 44 is enabled, thereby providing a sufficient signal to noise ratio. Also, longer fluorescence lifetime translates into larger integrated fluorescence photon count.
The fluorophores 246 (see Figure 59) have a fluorescence lifetime greater than 100 nanoseconds, often greater than 200 nanoseconds, more commonly greater than 300 nanoseconds and in most cases greater than 400 nanoseconds.
The metal-ligand complexes based on the transition metals or lanthanides have long lifetimes (from hundreds of nanoseconds to milliseconds), adequate quantum yields, and high thermal, chemical and photochemical stability, which are all favourable properties with respect to the fluorescence detection system requirements.
A particularly well-studied metal-ligand complex based on the transition metal ion Ruthenium (Ru (II)) is tris(2,2'-bipyridine) ruthenium (II) ([Ru(bpy)3]2+) which has a lifetime of approximately 1μ8. This complex is available commercially from Biosearch Technologies under the brand name Pulsar 650.
Table 1 : Photophysical properties of Pulsar 650 (Ruthenium chelate)
Terbium chelate, a lanthanide metal-ligand complex has been successfully demonstrated as a fluorescent reporter in a FRET probe system, and also has a Ion:
lifetime of 1600 8.
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Table 2: Photophysical properties of terbium chelate
The fluorescence detection system used by the LOC device 301 does not utilize filters to remove unwanted background fluorescence. It is therefore advantageous if the quencher 248 has no native emission in order to increase the signal-to-noise ratio. With no native emission, there is no contribution to background fluorescence from the quencher 248. High quenching efficiency is also important so that fluorescence is prevented until a hybridization event occurs. The Black Hole Quenchers (BHQ), available from Biosearch Technologies, Inc. of Novato California, have no native emission and high quenching efficiency, and are suitable quenchers for the system. BHQ- 1 has an absorption maximum at 534 nm, and a quenching range of 480-580 nm, making it a suitable quencher for the Tb- chelate fluorophore. BHQ-2 has an absorption maximum at 579 nm, and a quenching range of 560-670 nm, making it a suitable quencher for Pulsar 650.
Iowa Black Quenchers (Iowa Black FQ and RQ), available from Integrated DNA
Technologies of Coralville, Iowa, are suitable alternative quenchers with little or no background emission. Iowa Black FQ has a quenching range from 420-620 nm, with an absorption maximum at 531 nm and would therefore be a suitable quencher for the Tb- chelate fluorophore. Iowa Black RQ has an absorption maximum at 656 nm, and a quenching range of 500-700 nm, making it an ideal quencher for Pulsar 650.
In the embodiments described here, the quencher 248 is a functional moiety which is initially attached to the probe, but other embodiments are possible in which the quencher is a separate molecule free in solution.
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EXCITATION SOURCE
In the fluorescence detection based embodiments described herein, a LED is chosen as the excitation source instead of a laser diode, high power lamp or laser due to the low power consumption, low cost and small size. Referring to Figure 88, the LED 26 is positioned directly above the hybridization chamber array 110 on an external surface of the LOC device 301. On the opposing side of the hybridization chamber array 110, is the photosensor 44, made up of an array of photodiodes 184 (see Figures 53, 54 and 68) for detection of fluorescence signals from each of the chambers.
Figures 89, 90 and 91 schematically illustrate other embodiments for exposing the probes to excitation light. In the LOC device 30 shown in Figure 89, the excitation light 244 generated by the excitation LED 26 is directed onto the hybridization chamber array 1 10 by the lens 254. The excitation LED 26 is pulsed and the fluorescence emissions are detected by the photosensor 44.
In the LOC device 30 shown in Figure 90, the excitation light 244 generated by the excitation LED 26 is directed onto the hybridization chamber array 1 10 by the lens 254, a first optical prism 712 and second optical prism 714. The excitation LED 26 is pulsed and the fluorescence emissions are detected by the photosensor 44.
Similarly, the LOC device 30 shown in Figure 91, the excitation light 244 generated by the excitation LED 26 is directed onto the hybridization chamber array 1 10 by the lens 254, a first mirror 716 and second mirror 718. Again, the excitation LED 26 is pulsed and the fluorescence emissions are detected by the photosensor 44.
The excitation wavelength of the LED 26 is dependent on the choice of fluorescent dye. The Philips LX 2-PR14-R00 is a suitable excitation source for the Pulsar 650 dye. The SET UVTOP335T039BL LED is a suitable excitation source for the Tb-chelate label.
Table 3: Philips LXK2-PR14-R00 LED specifications
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Table 4: SET UVTOP334T039BL LED Specifications
ULTRA VIOLET EXCITATION LIGHT
Silicon absorbs little light in the UV spectrum. Accordingly, it is advantageous to use UV excitation light. A UV LED excitation source can be used but the broad spectrum of the LED 26 reduces the effectiveness of this method. To address this, a filtered UV LED can be used. Optionally, a UV laser can be the excitation source unless the relatively high cost of the laser is impractical for the particular test module market.
LED DRIVER
The LED driver 29 drives the LED 26 at a constant current for the required duration. A lower power USB 2.0-certifiable device can draw at most 1 unit load (100 mA), with a minimum operating voltage of 4.4 V. A standard power conditioning circuit is used for this purpose.
PHOTODIODE
Figure 54 shows the photodiode 184 integrated into the CMOS circuitry 86 of the LOC device 301. The photodiode 184 is fabricated as part of the CMOS circuitry 86 without additional masks or steps. This is one significant advantage of a CMOS photodiode over a CCD, an alternate sensing technology which could be integrated on the same chip using non-standard processing steps, or fabricated on an adjacent chip. On-chip detection is low cost and reduces the size of the assay system. The shorter optical path length reduces noise from the surrounding environment for efficient collection of the fluorescence signal and eliminates the need for a conventional optical assembly of lenses and filters.
Quantum efficiency of the photodiode 184 is the fraction of photons impinging on its active area 185 that are effectively converted to photo-electrons. For standard silicon
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processes, the quantum efficiency is in the range of 0.3 to 0.5 for visible light, depending on process parameters such as the amount and absorption properties of the cover layers.
The detection threshold of the photodiode 184 determines the smallest intensity of the fluorescence signal that can be detected. The detection threshold also determines the size of the photodiode 184 and hence the number of hybridization chambers 180 in the hybridization and detection section 52 (see Figure 52). The size and number of chambers are technical parameters that are limited by the dimensions of the LOC device (in the case of the LOC device 301, the dimensions are 1760 μηι x 5824 μηι) and the real estate available after other functional modules such as the pathogen dialysis section 70 and amplification section(s) 1 12 are incorporated.
For standard silicon processes, the photodiode 184 detects a minimum of 5 photons. However, to ensure reliable detection, the minimum can be set to 10 photons. Therefore with the quantum efficiency range being 0.3 to 0.5 (as discussed above), the fluorescence emission from the probes should be a minimum of 17 photons but 30 photons would incorporate a suitable margin of error for reliable detection.
CALIBRATION CHAMBERS
The non-uniformity of the electrical characteristic of the photodiode 184, autofluorescence, and residual excitation photon flux that has not yet completely decayed, introduce background noise and offset into the output signal. This background is removed from each output signal using one or more calibration signals. Calibration signals are generated by exposing one or more calibration photodiodes 184 in the array to respective calibration sources. A low calibration source is used for determining a negative result in which a target has not reacted with a probe. A high calibration source is indicative of a positive result from a probe-target complex. In the embodiment described here, the low calibration light source is provided by calibration chambers 382 in the hybridization chamber array 1 10 which:
do not contain any probes;
contain probes that have no fluorescent reporter; or,
contain probes with a reporter and quencher configured such that quenching is always expected to occur.
The output signal from such calibration chambers 382 closely approximates the noise and offset in the output signal from all the hybridization chambers in the LOC
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device. Subtracting the calibration signal from the output signals generated by the other hybridization chambers substantially removes the background and leaves the signal generated by the fluorescence emission (if any). Signals arising from ambient light in the region of the chamber array are also subtracted.
It will be appreciated that the negative control probes described above with reference to Figures 100 to 103 can be be used in calibration chambers. However, as shown in Figures 94 and 95, which are enlarged views of insets DG and DH of LOC variant X 728 shown in Figure 93, another option is to fluidically isolate the calibration chambers 382 from the amplicon. The background noise and offset can be determined by leaving the fluidically isolated chambers empty, or containing reporterless probes, or indeed any of the 'normal' probes with both reporter and quencher as hybridization is precluded by fluidic isolation.
The calibration chambers 382 can provide a high calibration source to generate a high signal in the corresponding photodiodes. The high signal corresponds to all probes in a chamber having hybridized. Spotting probes with reporters and no quenchers, or just reporters will consistently provide a signal approximating that of a hybridization chamber in which a predominant number of the probes have hybridized. It will also be appreciated that calibration chambers 382 can be used instead of control probes, or in addition to control probes.
The number and arrangement of the calibration chambers 382 throughout the hybridization chamber array is arbitrary. However, the calibration is more accurate if photodiodes 184 are calibrated by a calibration chamber 382 that is relatively proximate. Referring to Figure 56, the hybridization chamber array 110 has one calibration chamber 382 for every eight hybridization chambers 180. That is, a calibration chamber 382 is positioned in the middle of every three by three square of hybridization chambers 180. In this configuration, the hybridization chambers 180 are calibrated by a calibration chamber 382 that is immediately adjacent.
Figure 99 shows a differential imager circuit 788 used to substract the signal from the photodiode 184 corresponding to the calibration chamber 382 as a result of excitation light, from the fluorescence signal from the surrounding hybridization chambers 180. The differential imager circuit 788 samples the signal from the pixel 790 and a "dummy" pixel 792. In one embodiment, the "dummy" pixel 792 is shielded from light, so its output signal provides a dark reference. Alternatively, the "dummy" pixel 792 can be exposed to the excitation light along with the rest of the array. In the embodiment where the
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"dummy" pixel 792 is open to light, signals arising from ambient light in the region of the chamber array are also subtracted.The signals from the pixel 790 are small (i.e. close to dark signal), and without a reference to a dark level it is hard to differentiate between the background and a very small signal.
During use, the "read_row" 794 and "read row d" 795 are activated and M4 797 and MD4 801 transistors are turned on. Switches 807 and 809 are closed such that the outputs from the pixel 790 and "dummy" pixel 792 are stored on pixel capacitor 803 and dummy pixel capacitor 805 respectively. After the pixel signals have been stored, switches 807 and 809 are deactivated. Then the "read_col" switch 81 1 and dummy "read col" switch 813 are closed, and the switched capacitor amplifier 815 at the output amplifies the differential signal 817.
SUPPRESSION AND ENABLEMENT OF THE PHOTODIODE
The photodiode 184 needs to be suppressed during excitation by the LED 26 and enabled during fluorescence. Figure 69 is a circuit diagram for a single photodiode 184 and Figure 70 is a timing diagram for the photodiode control signals. The circuit has photodiode 184 and six MOS transistors, Mshunt 394, M^ 396, Mreset 398, Msf 400, Mread 402 and Mbias 404. At the beginning of the excitation cycle, tl, the transistors Mshunt 394, and Mreset 398 are turned on by pulling the MsriUnt gate 384 and the reset gate 388 high. During this period, the excitation photons generate carriers in the photodiode 184. These carriers have to be removed, as the amount of generated carriers can be sufficient to saturate the photodiode 184. During this cycle, Mshunt 394 directly removes the carriers generated in photodiode 184, while MreSet 398 resets any carriers that have accumulated on node 'NS' 406 due to leakage in transistors or due to diffusion of excitation-produced carriers in the substrate. After excitation, a capture cycle commences at t4. During this cycle, the emitted response from the fluorophore is captured and integrated in the circuit on node 'NS' 406. This is achieved by pulling tx gate 386 high, which turns on the transistor Mtx 396 and transfers any accumulated carriers on the photodiode 184 to node 'NS' 406. The duration of the capture cycle can be as long as the fluorophore emits. The outputs from all photodiodes 184 in the hybridization chamber array 110 are captured
simultaneously.
There is a delay between the end of the capture cycle t5 and the start of the read cycle t6. This delay is due to the requirement to read each photodiode 184 in the hybridization chamber array 110 (see Figure 52) separately following the capture cycle.
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The first photodiode 184 to be read will have the shortest delay before the read cycle, while the last photodiode 184 will have the longest delay before the read cycle. During the read cycle, transistor Mread 402 is turned on by pulling the read gate 393 high. The 'NS' node 406 voltage is buffered and read out using the source-follower transistor Msf 400.
There are additional, optional methods of enabling or suppressing the photodiode as discussed below:
1. Suppression Methods
Figures 96, 97 and 98 show three possible configurations 778, 780, 782 for the Mshunt transistor 394. The Mshunt transistor 394 has a very high off ratio at maximum
5 V which is enabled during excitation. As shown in Figure 96, the Mshunt gate 384 is configured to be on the edge of the photodiode 184. Optionally, as shown in Figure 97, the Mshunt gate 384 may be configured to surround the photodiode 184. A third option is to configure the Msnunt gate 384 inside the photodiode 184, as shown in Figure 98. Under this third option there would be less photodiode active area 185.
These three configurations 778, 780 and 782 reduce the average path length from all locations in the photodiode 184 to the Mshunt gate 384. In Figure 96, the Mshunt gate 384 is on one side of the photodiode 184. This configuration is simplest to fabricate and impinges the least on the photodiode active area 185. However, any carriers lingering on the remote side of the photodiode 184 would take longer to propagate through to the Mshunt gate 384.
In Figure 97, the Mshunt gate 384 surrounds the photodiode 184. This further reduces the average path length for carriers in the photodiode 184 to the Mshunt gate 384. However, extending the Mshunt gate 384 about the periphery of the photodiode 184 imposes a greater reduction of the photodiode active area 185. The configuration 782 in Figure 98 positions the Mshunt gate 384 within the active area 185. This provides the shortest average path length to the Mshunt gate 384 and hence the shortest transition time. However, the impingement on the active area 185 is greatest. It also poses a wider leakage path.
2. Enabling Methods
a. A trigger photodiode drives the shunt transistor with a fixed delay.
b. A trigger photodiode drives the shunt transistor with programmable delay.
c. The shunt transistor is driven from the LED drive pulse with a fixed delay.
d. The shunt transistor is driven as in 2c but with programmable delay.
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Figure 75 is a schematic section view through a hybridization chamber 180 showing a photodiode 184 and trigger photodiode 187 embedded in the CMOS circuitry 86. A small area in the corner of the photodiode 184 is replaced with the trigger photodiode 187. A trigger photodiode 187 with a small area is sufficient as the intensity of the excitation light will be high in comparison with the fluorescence emission. The trigger photodiode 187 is sensitive to the excitation light 244. The trigger photodiode 187 registers that the excitation light 244 has extinguished and activates the photodiode 184 after a short time delay At 300 (see Figure 2). This delay allows the fluorescence photodiode 184 to detect the fluorescence emission from the FRET probes 186 in the absence of the excitation light 244. This enables detection and improves the signal to noise ratio.
Both photodiodes 184 and trigger photodiodes 187 are located in the CMOS circuitry 86 under each hybridization chamber 180. The array of photodiodes combines, along with appropriate electronics, to form the photosensor 44 (see Figure 68). The photodiodes 184 are pn-junction fabricated during CMOS structure manufacturing without additional masks or steps. During MST fabrication, the dielectric layer (not shown) above the photodiodes 184 is optionally thinned using the standard MST photolithography techniques to allow more fluorescent light to illuminate the active area 185 of the photodiode 184. The photodiode 184 has a field of view such that the fluorescence signal from the probe-target hybrids within the hybridization chamber 180 is incident on the sensor face. The fluorescent light is converted into a photocurrent which can then be measured using CMOS circuitry 86.
Alternatively, one or more hybridization chambers 180 can be dedicated to a trigger photodiode 187 only. These options can be used in these in combination with 2a and 2b above.
DELAYED DETECTION OF FLUORESCENCE
The following derivations elucidate the delayed detection of fluorescence using a long-lifetime fluorophore for the LED/fluorophore combinations described above. The fluorescence intensity is derived as a function of time after excitation by an ideal pulse of constant intensity Ie between time t\ and ¾ as shown in Figure 60.
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Let [S1]( equal the density of excited states at time t, then during and after excitation, the number of excited states per unit time per unit volume is described by the following differential equation:
^(t)+ sm = i c_ ( 1)
dt Tf h ve
where c is the molar concentration of fluorophores, ε is the molar extinction coefficient, ve is the excitation frequency, and h = 6.62606896(10)~34 Js is the Planck constant.
This differential equation has the general form:
^ + p(x)y = q(x)
dx
which has the solution:
C \ p(x)dx
e' q(x)dx + k
y(x) = - r— ...(2)
p{x)dx
Using this now to solve equation (1),
\Sl](t) ^ -tl'x
ke r,
.(3)
h ve
Now at time h, [Sl](h) = 0, and from (3): k = -—— Leh '-f ...(4)
h ve
Substituting (4) into (3): h ve h ve
For t > t2, the excited states decay exponentially and this is described by:
[Sl](t) = [Sl](t2 )e~(t~'2)lTf ...(6)
The fluorescence intensity is given by the following equation:
d[Sl](t) ,
if (t) = - d^ h vf1il - - -(8)
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where V/is the fluorescence frequency, η is the quantum yield and 1 is the optical path length.
Substitutin (9) into (8):
If it) = - e h~h),Ti y'~h Tf ...(10)
For iL→ oo, I (ί)→Ιεεάη^β~{
Therefore, we can write the following approximate equation which describes the fluorescence intensity decay after a sufficiently long excitation pulse (¾-ti » if):
In the previous section, we concluded that for ti-h » Tf, lM) = lescl ^e (,~ )lT* for t≥t2.
From the above equation, we can derive the following:
ηΜ) = ηβ8οΙηβ ,~,ι)!τί ...(12)
where
I fi)
nf(t) =—— is the number of fluorescent photons per unit time per unit area and
hvf n = -^- is the number of excitation photons per unit time per unit area.
hve
Consequently, nf(t) = nf(t)dt ...(13)
h
where nr is the number of fluorescent photons per unit area and ?3 is the instant of time at which the photodiode is turned on. Substituting (12) into (13): εοΙηβ ('~ )'τι dt ...(14)
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Now, the number of fluorescent photons that reach the photodiode per unit time per unit area, ns (t) , is given by the following:
where φ0 is the light gathering efficiency of the optical system.
Substituting (12) into (15) we find
η, (ί) = Φ0ηβεάψ-('-'ι τ/ ...(16)
Similarly, the number of fluorescence photons that reach the photodiode per unit fluorescent area ns , will be as follows: hs = j" ns (t)dt and substituting in (16) and integrating:
h
Therefore,
ns = φ0ήεεοΙητ fe~ lTf - - -(17)
The optimal value of ¾ is when the rate of electrons generated in the photodiode 184 due to fluorescence photons becomes equal to the rate of electrons generated in the photodiode 184 by the excitation photons, as the flux of the excitation photons decays much faster than that of the fluorescence photons.
where φ/ is the quantum efficiency of the sensor at the fluorescence wavelength.
Substituting in (17) we have:
βί (ΐ) = φίφ -η€εοΙηβ-(,-Η )ίτ' ...(18)
Similarly, the rate of sensor output electrons per unit fluorescent area due to the excitation photons is:
¾( = ^¾ ("2)/Te •••(19)
where φ is the quantum efficiency of the sensor at the excitation wavelength, and xe is the time-constant corresponding to the "off characteristics of the excitation LED. After time t2, the LED's decaying photon flux would increase the intensity of the fluorescence signal and extend its decay time, but we are assuming that this has a negligible effect on ¾{t), thus we are taking a conservative approach.
GCA003-PCT
Now, as mentioned earlier, the optimal value of t3 is when:
ef (t3) = e (t} )
where F = εεΙη and At = t3 - t2. We also know that, in practice, t2 - tx » τ f .
The optimal time for fluorescence detection and the number of fluorescence photons detected using the Philips LXK2-PR14-R00 LED and Pulsar 650 dye are determined as follows.
The optimum detection time is determined using equation (22):
Recalling the concentration of amplicon, and assuming that all amplicons hybridize, then the concentration of fluorescent fluorophores is: c = 2.89(10)"6 mol/L
The height of the chamber is the optical path length 1 = 8(10)~6 m.
We have taken the fluorescence area to be equal to our photodiode area, yet our actual fluorescence area is substantially larger than our photodiode area; consequently we can approximately assume φ0 = 0.5 for the light gathering efficiency of our optical system.
From the photodiode characteristics,— = 10 is a very conservative value for the ratio of the photodiode quantum efficiency at the fluorescence wavelength to its quantum efficiency at the excitation wavelength.
With a typical LED decay lifetime of τε = 0.5 ns and using Pulsar 650
specifications, At can be determined:
GCA003-PCT
F = [1.48(10)6 ][2.89(10) 6 ][8(10) 6 ](1)
= 3.42(10)"5
= 4.34(10)"9 s
The number of photons detected is determined using equation (21). First, the number of excitation photons emitted per unit time he is determined by examining the illumination geometry.
The Philips LXK2-PR14-R00 LED has a Lambertian radiation pattern, therefore: j¾ = ¾f/0 cos(0) ...(23) where n, is the number of photons emitted per unit time per unit solid angle at an angle of Θ off the LED's forward axial direction, and nm is the valve of n) in the forward axial direction.
The total number of photons emitted by the LED per unit time is:
Now,
ΔΩ 2 I - cos(# + Δ0)] - 2 I - cos(0)]
άΩ. = 2πήη(θ)άθ
= m Ί.0
Rearranging, we have:
GCA003-PCT
¾„ = - •••(26)
π
The LED's output power is 0.515 W and ve = 6.52(10)14 Hz, therefore: n, = -p- -(27)
h ve
_ 0.515
" [6.63(10Γ34][6.52(10)14]
= 1.19(10)18 photons/s
Substituting this value into (26) we have:
... 1.19(10)18
¾ =
π
= 3.79(10)17 photons/s/sr
Referring to Figure 61, the optical centre 252 and the lens 254 of the LED 26 are schematically shown. The photodiodes are 16 μιη x 16 μηι, and for the photodiode in the middle of the array, the solid angle (Ω) of the cone of light emitted from the LED 26 to the photodiode 184 is approximately:
Ω = area of sensor/r2
_ [16(10) 6 ][16(10) 6 ]
[2.825(10) 3f
= 3.21(10)"5 sr
It will be appreciated that the central photodiode 184 of the photodiode array 44 is used for the purpose of these calculations. A sensor located at the edge of the array would only receive 2% less photons upon a hybridization event for a Lambertian excitation source intensity distribution.
The number of excitation photons emitted per unit time is:
ήβ = η,Ω ...(28)
= [3.79(10)17][3.21(1 O 5]
= 1.22(10)13 photons/s
Now referring to equation (29):
ns = (/>0neFTfe ns = (0.5)[1.22(10)13 ][3.42(10)-5 ][l(10)-6 ]e^34(10r'/,(1()r6
= 208 photons per sensor.
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Therefore, using the Philips LXK2-PR14-R00 LED and Pulsar 650 fluorophore, we can easily detect any hybridization events which results in this number of photons being emitted.
The SET LED illumination geometry is shown in Figure 62. At ID = 20 iriA, the LED has a minimum optical power output of pi = 240 μW centred at λβ = 340 nm (the absorption wavelength of the terbium chelate). Driving the LED at ID = 200 mA would increase the output power linearly to pi = 2.4 mW. By placing the LED's optical centre 252, 17.5 mm away from the hybridization chamber array 110, we would approximately concentrate this output flux in a circular spot size which has a maximum diameter of 2 mm.
2.4(10Γ3
[6.63(10r34 ][8.82(10)14 ]
= 4.10(10)15 photons/s
Using equation 28, we have:
K = η,Ω
= 4.10(10)15 [l6^ 2
= 3.34(10)u photons/s
Now, recalling equation 22 and using the Tb chelate properties listed previously, ln[(6.94(10r5 )(10)(0.5)]
At
1 1
1(10) 3 0.5(10Γ9
= 3.98(10)"9 s
Now from equation 21 :
ns = (0.5)[3.34(10)n ][6.94(10)-5 ][l(10)-3 ]e-3-98(10r' 1(10
= 11 ,600 photons per sensor.
GCA003-PCT
The theoretical number of photons emitted by hybridization events using the SET LED and terbium chelate system are easily detectable and well over the minimum of 30 photons required for reliable detection by the photosensor as indicated above.
MAXIMUM SPACING BETWEEN PROBES AND PHOTODIODE The on-chip detection of hybridization avoids the needs for detection via confocal microscopy (see Background of the Invention). This departure from traditional detection techniques is a significant factor in the time and cost savings associated with this system. Traditional detection requires imaging optics which necessarily uses lenses or curved mirrors. By adopting non-imaging optics, the diagnostic system avoids the need for a complex and bulky optical train. Positioning the photodiode very close to the probes has the advantage of extremely high collection efficiency: when the thickness of the material between the probes and the photodiode is of the order of 1 micron, the angle of collection of emission light is up to 173°. This angle is calculated by considering light emitted from a probe at the centroid of the face of the hybridization chamber closest to the photodiode, which has a planar active surface area parallel to that chamber face. The cone of emission angles within which light is able to be absorbed by the photodiode is defined as having the emitting probe at its vertex and the corner of the sensor on the perimeter of its planar face. For a 16 micron x 16 micron sensor, the vertex angle of this cone is 170°; in the limiting case where the photodiode is expanded so that its area matches that of the 29 micron x 19.75 micron hybridization chamber, the vertex angle is 173°. A separation between the chamber face and the photodiode active surface of 1 micron or less is readily achievable.
Employing a non-imaging optics scheme does require the photodiode 184 to be very close to the hybridization chamber in order to collect sufficient photons of fluorescence emission. The maximum spacing between the photodiode and probes is determined as follows with reference to Figure 54.
Utilizing a terbium chelate fluorophore and a SET UVTOP335T039BL LED, we calculated 11600 photons reaching our 16 micron x 16 micron photodiode 184 from the respective hybridization chamber 180. In performing this calculation we assumed that the light-collecting region of our hybridization chamber 180 has a base area which is the same as our photodiode active area 185, and half of the total number of the hybridization photons reaches the photodiode 184. That is, the light gathering efficiency of the optical system is φ0 = 0.5 .
GCA003-PCT
More accurately we can write 0 = [(base area of the light-collecting region of the hybridization chamber)/(photodiode area)][Q/½], where Ω = solid angle subtended by the photodiode at a representative point on the base of the hybridization chamber. For a right square pyramid geometry:
Ω = 4arcsin(a2/(4do2+a2)), where do = distance between the chamber and the photodiode, and a is the photodiode dimension.
Each hybridization chamber releases 23200 photons. The selected photodiode has a detection threshold of 17 photons; therefore, the minimum optical efficiency required is: φ0= 17/23200 = 7.33xl0"4
The base area of the light-collecting region of the hybridization chamber 180 is 29 micron x 19.75 micron.
Solving for do, we will get the maximum limiting distance between the bottom of our hybridization chamber and our photodiode 184 to be do = 249 microns. In this limit, the collection cone angle as defined above is only 0.8°. It should be noted this analysis ignores the negligible effect of refraction.
TEST MODULE WITH MICROFLUIDIC DEVICE HAVING DIALYSIS DEVICE, LOC AND INTERCONNECTING CAP
A test module 11 for analysing a sample fluid containing target molecules is shown in Figure 109. The test module 11 comprises an outer casing 13 with a receptacle 24 for receiving the sample fluid, a removable sterile sealing tape 22 to cover the receptacle 24 prior to use, a membrane seal 408 with a membrane guard 410 forming part of the outer casing 13 to reduce dehumidification within the test module while providing pressure relief from small air pressure fluctuations with the membrane guard 410 protecting the membrane seal 408 from damage, a printed circuit board (PCB) 57, a microfluidic device 783, a porous element 49, a standard Micro-USB plug 14 for power, data and control, external power supply capacitors 32, and inductor 15.
The microfluidic device 783 has a dialysis device 784 in fluid communication with the receptacle 24 and configured to separate the target molecules from other constituents of the sample, a LOC device 785 for analysing the target molecules and a cap 51 overlaying the LOC device 785 and the dialysis device 784 for establishing fluid communication between the LOC device 785 and the dialysis device 784.
REAGENT LOADING AND PROBE SPOTTING SYSTEM
GCA003-PCT
Reagent reservoirs 54, 56, 58, 60 and 62 (see Figure 6) are filled with reagents and water from a robotic, droplet ejection system shown in Figures 63 to 66. The robotic system also spots the oligonucleotide FRET probes 186 or ECL probes 237 into the hybridization chambers 180. Droplet dispensing technology is an inexpensive spotting technique, delivers small droplets with reproducible volumes and many droplets of different solutions can be dispensed simultaneously. This allows the LOC devices to be mass produced at extremely high throughput and low cost.
The reagent and probe spotting system includes three robotic subsystems:
1. Reagent dispensing robot 256 (see Figure 63) - microvials 258 (see Figure 64), each with a droplet dispenser 262, dispense reagents into the reservoirs 54, 56, 58, 60 and
62 and water into the water reservoir 188 (see Figure 6). It then applies the patterned upper seal 82 (if necessary) to the cap 46.
2. ONEC refill robot 274 (see Figure 65) - microvials 258 with a droplet dispenser 262 dispense probes into the reservoirs 278 of an oligonucleotide ejector chip (ONEC) 272 (see Figures 71 and 72). The ONEC reservoirs 278 feed an array of thermal droplet generators 271. The ONEC is then used in the third robotic subsystem, the LOC spotting robot.
3. LOC spotting robot 289 (schematically shown in Figure 66) - ONEC 272 spots each hybridization chamber 180 of the LOC device 30 with probes using a thermal droplet generator 271 (see Figure 72).
MICROVIALS
The reagent dispensing robot 256 and the ONEC refill robot 274 both use microvials 2 8 as shown schematically in Figure 64. Probes and reagents are ordered directly from the suppliers in macrovials (not shown). Liquids are micropipetted from the macrovials into a container 259 on each of the microvials 258 to form small aliquots
(typically between 282 microliters and 400 microliters) that can be refrigerated along with the macrovials until required. Each microvial 258 has a piezoelectric droplet dispenser 262 and an enclosed quality assurance chip (i.e. integrated circuit) 266 with flash memory and electrical contacts 264 for power and data transmission. The droplet dispenser 262 has a piezo-electric actuator 261 configured to eject drops with a volume between 50 picoliters and 150 picoliters for reasonably quick reagent loading while maintaining accurate drop placement.
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PROBE AND REAGENT IDENTIFICATION SCHEME
The quality assurance chip 266 (see Figure 64) has digital memory used to store, identify and track the specification data characterizing the reagent or oligonucleotide probe solution within the microvial 258. At the end of the spot and load process, the data from each microvial 258, along with other loading and spotting data, is downloaded and stored in the program and data flash memory 40 of the LOC device 30 via the control microprocessor 263 controlling the reagent dispensing robot or probe dispensing robot. This data is used for diagnostic information and processing tasks, quality control and auditing.
Referring to Figure 73, ONEC 272 also has digital memory such as flash memory 281 in the ONEC CMOS structure 285 to store oligonucleotide specification data such as probe identities, batch numbers and so on. As with the LOC device, the ONEC refill robot 274 downloads the specification data to the ONEC flash memory 281 from the quality assurance chips 266 on the microvials 258.
Automated information transfer minimizes the possibility of errors occurring and in the event an incorrect microvial is used, the test module reader 12 or other system component identifies this error when processing the diagnostic information.
REAGENT DISPENSING ROBOT
A simplified top and side view of the reagent dispensing robot 256 are shown in Figures 63 and 108. It includes:
• microvials 258 containing reagents and molecular biology grade water (only some of the microvials are shown)
• mechanical/electrical rack 286 (shown only in outline) which holds and provides electrical connectivity to microvials 258
• XY stage 268 providing a surface for detachably mounting a partial-depth sawn silicon wafer 260 or other fixed array such as separable PCB wafer 720
• Registration camera 270 providing feedback to the control microprocessor 263 for mapping the exact location of the piezoelectric droplet dispensers 262
GCA003-PCT
The piezoelectric droplet dispensers 262 on the microvials 258 are used to dispense the reagents and water directly into the LOG device reservoirs 54, 56, 58, 60 and 62 and the humidifier water reservoir 188 respectively.
ONEC REFILL ROBOT
The ONEC refill robot 274 is shown in Figure 65. It is similar to the reagent dispensing robot 256 and includes:
• 1080 microvials 258 containing solutions of oligonucleotide probes (for the
purposes of illustration, not all microvials are shown)
• mechanical/electrical rack 286 (shown only in outline) - holds and provides
electrical connectivity to microvials 258
• oligonucleotide ejector chip (ONEC) 272 - with 1080 ONEC reservoirs 278
supplying respective ejectors 287 with four ONEC thermal droplet generators 271 each (see Figures 71 and 72)
• XY stage 268: holds the oligonucleotide ejector chip/s (ONEC/s) 272
• Registration camera 270 providing feedback to the control microprocessor 263 for mapping the exact location of the thermal droplet generators 271
The ONEC 272 is moved under the mechanical/electrical rack 286. A unique probe solution is dispensed from each microvial 258 into each ONEC reservoir 278. The ONEC 272 is then used in the probe spotting robot 273 to spot the LOC device hybridization chambers 180 with a single droplet of probe solution.
ONEC
Figures 71, 72 and 73 show the ONEC 272 in detail. The ONEC 272 is an oligonucleotide spotting device for contactless spotting of probes onto a surface such as the hybridization chamber array in any of the LOC devices. It has overall dimensions of 23,296 μιη x 1 ,760 μιη and is fabricated using well-established high volume
photolithography fabrication techniques. Each ONEC has 1080 reservoirs 278 etched into the reservoir side 277 of a monolithic silicon substrate 275 (see Figure 73). With more than 1000 reservoirs 278, each ONEC has the complete assay of probes needed to spot the LOC devices described herein. This allows the spotting process of each LOC to be one- step in the sense that there is no need to use more than one ONEC to spot LOCs configured
GCA003-PCT
for each particular analysis. The ONEC reservoirs 278 have a rectangular base (96 μηι x 208 μηι) with a depth of 200 μηχ Each ONEC reservoir 278 feeds a probe suspension to a respective ejector 287. The liquid suspension of probes fill a common chamber 282 via a pair of chamber inlets 284 (see Figure 72). The chamber inlets 284 are two 21 μιη diameter holes from the reservoir 278 to the common chamber 282. One of four thermal droplet generators 271 ejects probe droplets through nozzles 283 in the ejector side 279 into the hybridization chambers 180 by heating the actuator 280 to generate a vapor bubble. Having four thermal droplet generators 271 allows for redundancy if there is a droplet generator failure. LOC PROBE SPOTTING ROBOT
The LOC probe spotting robot 289 is shown in Figures 66 and 92. For clarity, components other than the LOC device 30 on the PCB wafer 720 are not shown. It includes the following:
• ONEC 272 - oligonucleotide ejector chip with 1080 reservoirs 278, each filled with a probe solution (see Figures 71 and 72)
• XY stage 268: holds the partial-depth sawn silicon LOC wafer 260 (see Figure 66) or alternatively the separable PCB wafer 720 (see Figure 92)
• Registration camera 270 providing feedback to the control processor 263 for
mapping the exact location of the ONEC thermal droplet generators 271
The LOC silicon wafer 260 or the separable PCB wafer 720 is detachably mounted to a stage that can translate along two orthogonal axes. The ONEC 272 is detachably held in a chuck 265 that is closely adjacent the stage with the ejectors 287 facing the stage (see Figure 66). The LOC silicon wafer 260 or the separable PCB wafer 720 is moved relative to the ONEC 272 by the control processor 263. Each LOC device hybridization chamber 180 is spotted by the ejectors under the operative control of the control processor 263. Using volumes less than 100 picoliters reduces the reaction times and allows the density of the hybridization chamber array to increase. Spotting low-volume probe droplets has not been previously adopted because of the difficulty associated with ejecting very small droplets precisely and reliably. Misdirected drops can fail to spot the correct chamber and may contaminate an adjacent chamber.
GCA003-PCT
The ONEC 272 can be driven to generate a range of droplet volumes. For accurate dispensing, the droplets generated by the ONEC 272 would be less than 100 picoliters. To improve the accuracy of the probes and reagents dispensed (in terms of volume and position on the LOC device), the droplets generated by the ONEC can be reduced to less than 25 picoliters, and preferably less than 6 picoliters. The ONEC 272 dispenses probe solution into the 1080 hybridization chambers 180 in droplets with volumes between 0.1 picoliters and 1.6 picoliters and a high degree of positional accuracy.
The hybridization chamber array 110 is configured as 24 rows with 45 adjacent chambers in each row (see Figure 52). The sample flow-path 176 extends between every second row such that the overall array has a substantially square shape for approximately uniform illumination by the LED 26. As the hybridization chamber array 110 is confined to an area less than 1500 microns by 1500 microns, the spotting accuracy of the ONEC 272 is necessarily high. A registration camera 270 is used by the control processor 263 to determine the exact position of the ONEC thermal droplet generators 271 and the droplet generator drive pulses are synchronized with the XY stage 268 via the ONEC bond-pads 276.
The LOC probe spotting robot 273 using the ONEC 272 and camera 270 can easily spot probes onto a surface (such as the hybridization chamber array 110) at a rate greater than 100 probes per second; in the vast majority of cases at a rate greater than 1 ,400 probes per second. Typically, the array of droplet generators spot the probes onto the surface at a rate greater than 20,000 probes per second and in many cases, the array of droplet generators spot the probes onto the surface at a rate between 300,000 probes per second and 1,000,000 probes per second.
The array of droplet generators lithographically fabricated on a silicon substrate allows the ONEC 272 to spot oligonucleotides onto a surface at a density far greater than existing probe spotters. ONEC 272 easily spots at a density of more than 1 probe per square millimetre. In the vast majority of cases, the spotting density is greater than 8 probes per square millimetre. In most cases, the spotting density is more than 60 probes per square millimetre, and typically the density is between 500 probes per square millimetre and 1,500 probes per square millimetre.
The LOC probe spotting robot 273, using the ONEC 272 as a biochemical deposition device, can easily deposit biochemicals onto a surface at a rate greater than 100 droplets per second, in the vast majority of cases at a rate greater than 1,400 droplets per second. Typically, the array of droplet generators spot the droplets onto the surface at a
GCA003-PCT
rate greater than 20,000 droplets per second, and in many cases, the array of droplet generators spot the droplets onto the surface at a rate between 300,000 droplets per second and 1,000,000 droplets per second.
The LOC probe spotting robot 273, using the ONEC 272 as a biochemical deposition device, can easily deposit biochemicals onto a surface at a density of more than 1 droplet per square millimetre. In the vast majority of cases, the spotting density is greater than 8 droplets per square millimetre. In most cases, the spotting density is more than 60 droplets per square millimetre, and typically the density is between 500 droplets per square millimetre and 1 ,500 droplets per square millimetre.
CONCLUSION
The devices, systems and methods described here facilitate molecular diagnostic tests at low cost with high speed and at the point-of-care.
The system and its components described above are purely illustrative and the skilled worker in this field will readily recognize many variations and modifications which do not depart from the spirit and scope of the broad inventive concept.
GCA003-PCT
Claims
1. An apparatus for loading oligonucleotide spotting devices and spotting
oligonucleotide probes, the apparatus comprising:
a plurality of oligonucleotide vials, each with a droplet dispenser;
a mounting surface for detachably mounting an oligonucleotide spotting device; a chuck for detachably mounting the oligonucleotide spotting device adjacent the mounting surface; and,
a control processor for operative control of the oligonucleotide vials, the oligonucleotide spotting device when mounted in the chuck and movement of the mounting surface relative to the oligonucleotide vials, and the oligonucleotide spotting device; wherein,
the control processor is configured to activate the droplet dispensers, and move the oligonucleotide spotting device into registration with the oligonucleotide vials.
2. The apparatus according to claim 1 wherein the control processor is configured to operate the oligonucleotide spotting device when in the chuck to spot oligonucleotide probes into a microfluidic device on the mounting surface.
3. The apparatus according to claim 1 wherein each of the oligonucleotide vials has an integrated circuit storing oligonucleotide specification data, and the control processor is configured to download the oligonucleotide specification data to digital memory in the oligonucleotide spotting device.
4. The apparatus according to claim 1 further comprising a camera for optical feedback of the registration between the vial selected by the control processor and the oligonucleotide spotting device.
5. The apparatus according to claim 3 wherein the oligonucleotide vials are microvials with a volume between 282 microliters and 400 microliters.
6. The apparatus according to claim 5 wherein the integrated circuit for each of the microvials has a unique identifier for identifying each of the microvials individually, the unique identifier being transmitted to the control processor.
GCA003-PCT
7. The apparatus according to claim 6 wherein each of the microvials has electrical contacts for receiving activation pulses for the droplet dispenser and allowing the control processor to interrogate the integrated circuit.
8. The apparatus according to claim 6 further comprising a rack wherein the microvials are detachably mounted to the rack for mechanical and electronic control of the microvials.
9. The apparatus according to claim 8 wherein the mounting surface is a stage configured for movement along two orthogonal axes, the rack extending parallel to one if the orthogonal axes.
10. The apparatus according to claim 1 wherein the droplet dispenser has a piezo- electric actuator.
1 1. The apparatus according to claim 1 wherein the droplet dispenser is configured to eject droplets with a volume between 50 picoliters and 150 picoliters.
12. The apparatus according to claim 2 further comprising reagent vials containing reagents for processing a biological sample wherein the microfluidic device is a LOC device for genetic analysis of the biological sample, the LOC device having a polymerase chain reaction (PC ) section and the list of reagents has one or more of:
water;
polymerase;
primers;
buffer;
anticoagulant;
deoxyribonucleoside triphosphates (dNTPs);
lysis reagent;
ligase and linkers; and,
restriction enzymes.
GCA003-PCT
13. The apparatus according to claim 12 further comprising a facility for applying a film to the LOC device to cover reagent reservoirs formed in an exterior surface.
14. The apparatus according to claim 12 wherein the LOC device is one of an array of LOC devices fabricated on a silicon wafer, the stage being configured to detachably mount the silicon wafer for loading reagents into all the LOC devices in the array.
15. The apparatus according to claim 12 wherein the LOC device is one of an array of LOC devices mounted on a printed circuit board (PCB), the stage being configured to detachably mount the PCB for loading reagents into all the LOC devices in the array.
16. The apparatus according to claim 2 wherein the oligonucleotide spotting device has an array of reservoirs for containing the oligonucleotide probes and an array of ejectors, and the LOC device has an array of hybridization chambers for receiving the
oligonucleotide probes, the probes having nucleic acid sequences that are complementary to target nucleic acid sequences to be identified in a biological sample and the array of hybridization chambers being configured to hold a complete assay of oligonucleotide probes necessary for a predetermined analysis of the biological sample, and the array of reservoirs is configured to contain the complete assay of oligonucleotide probes necessary for the predetermined analysis to be performed by the LOC device, the control processor being configured to operate the ejectors to correctly spot the hybridization chamber array and download an association between the specification data for the oligonucleotide probes from each of the reservoirs, and array location data locating the hybridization chamber spotted by each of the reservoirs.
17. The apparatus according to claim 16 wherein each of the ejectors has a plurality of nozzles, a chamber for containing liquid with a suspension of the oligonucleotide probes from the corresponding reservoir, and a plurality of actuators, one of the actuators corresponding to each of the nozzles respectively such that the actuator ejects a droplet of the liquid from the chamber through the corresponding nozzle, the control processor being configured to operate each of the actuators individually.
GCA003-PCT
18. The apparatus according to claim 17 wherein the control processor is configured to operate the array of ejectors to spot the oligonucleotides onto the LOC device with a density greater than 8 probe spots per square millimeter.
19. The apparatus according to claim 17 wherein the control processor is configured to operate the array of ejectors to spot the oligonucleotides onto the LOC device with a density greater than 60 probe spots per square millimeter.
20. The apparatus according to claim 17 wherein the control processor is configured to operate the array of ejectors to spot the oligonucleotides onto the LOC device with a density between 500 probe spots per square millimeter and 1500 probe spots per square millimeter.
GCA003-PCT
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US35601810P | 2010-06-17 | 2010-06-17 | |
US61/356,018 | 2010-06-17 | ||
US201161437686P | 2011-01-30 | 2011-01-30 | |
US61/437,686 | 2011-01-30 |
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WO2011156836A1 true WO2011156836A1 (en) | 2011-12-22 |
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PCT/AU2011/000662 WO2011156839A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for genetic analysis with dialysis, chemical lysis and tandem nucleic acid amplification |
PCT/AU2011/000668 WO2011156845A1 (en) | 2010-06-17 | 2011-06-01 | Loc device with parallel incubation and parallel dna and rna amplification functionality |
PCT/AU2011/000673 WO2011156850A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic device with pcr section and diffusion mixer |
PCT/AU2011/000671 WO2011156848A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic device with flow-channel structure having active valve for capillary-driven fluidic propulsion without trapped air bubbles |
PCT/AU2011/000665 WO2011156842A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for pathogen detection and genetic analysis with dialysis and nucleic acid amplification |
PCT/AU2011/000675 WO2011156852A1 (en) | 2010-06-17 | 2011-06-01 | Genetic analysis loc with non-specific nucleic acid amplification section and subsequent specific amplification of particular sequences in a separate section |
PCT/AU2011/000669 WO2011156846A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic device for simultaneous detection of multiple conditions in a patient |
PCT/AU2011/000674 WO2011156851A1 (en) | 2010-06-17 | 2011-06-01 | Test module with diffusive mixing in small cross section area microchannel |
PCT/AU2011/000677 WO2011156854A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic device with conductivity sensor |
PCT/AU2011/000659 WO2011156836A1 (en) | 2010-06-17 | 2011-06-01 | Apparatus for loading oligonucleotide spotting devices and spotting oligonucleotide probes |
PCT/AU2011/000670 WO2011156847A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic device for genetic and mitochondrial analysis of a biological sample |
PCT/AU2011/000661 WO2011156838A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for pathogen detection with dialysis, chemical lysis and tandem nucleic acid amplification |
PCT/AU2011/000678 WO2011156855A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic device with reagent mixing proportions determined by number of active outlet valves |
PCT/AU2011/000680 WO2011156857A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic device with dialysis section having stomata tapering counter to flow direction |
PCT/AU2011/000664 WO2011156841A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for pathogen detection with dialysis, thermal lysis, nucleic acid amplification and prehybridization filtering |
PCT/AU2011/000666 WO2011156843A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for genetic analysis which performs nucleic acid amplification after sample preparation in a dialysis section |
PCT/AU2011/000667 WO2011156844A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for genetic analysis which performs nucleic acid amplification before removing non-nucleic acid constituents in a dialysis section |
PCT/AU2011/000672 WO2011156849A1 (en) | 2010-06-17 | 2011-06-01 | Test module with microfluidic device having loc and dialysis device for separating pathogens from other constituents in a biological sample |
PCT/AU2011/000663 WO2011156840A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for pathogen detection and genetic analysis with chemical lysis, incubation and tandem nucleic acid amplification |
PCT/AU2011/000658 WO2011156835A1 (en) | 2010-06-17 | 2011-06-01 | Test module incorporating spectrometer |
PCT/AU2011/000660 WO2011156837A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for pathogen detection with dialysis, thermal lysis, nucleic acid amplification and prehybridization filtering |
PCT/AU2011/000679 WO2011156856A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic thermal bend actuated surface tension valve |
PCT/AU2011/000676 WO2011156853A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic test module with flexible membrane for internal microenvironment pressure-relief |
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PCT/AU2011/000662 WO2011156839A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for genetic analysis with dialysis, chemical lysis and tandem nucleic acid amplification |
PCT/AU2011/000668 WO2011156845A1 (en) | 2010-06-17 | 2011-06-01 | Loc device with parallel incubation and parallel dna and rna amplification functionality |
PCT/AU2011/000673 WO2011156850A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic device with pcr section and diffusion mixer |
PCT/AU2011/000671 WO2011156848A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic device with flow-channel structure having active valve for capillary-driven fluidic propulsion without trapped air bubbles |
PCT/AU2011/000665 WO2011156842A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for pathogen detection and genetic analysis with dialysis and nucleic acid amplification |
PCT/AU2011/000675 WO2011156852A1 (en) | 2010-06-17 | 2011-06-01 | Genetic analysis loc with non-specific nucleic acid amplification section and subsequent specific amplification of particular sequences in a separate section |
PCT/AU2011/000669 WO2011156846A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic device for simultaneous detection of multiple conditions in a patient |
PCT/AU2011/000674 WO2011156851A1 (en) | 2010-06-17 | 2011-06-01 | Test module with diffusive mixing in small cross section area microchannel |
PCT/AU2011/000677 WO2011156854A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic device with conductivity sensor |
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PCT/AU2011/000670 WO2011156847A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic device for genetic and mitochondrial analysis of a biological sample |
PCT/AU2011/000661 WO2011156838A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for pathogen detection with dialysis, chemical lysis and tandem nucleic acid amplification |
PCT/AU2011/000678 WO2011156855A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic device with reagent mixing proportions determined by number of active outlet valves |
PCT/AU2011/000680 WO2011156857A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic device with dialysis section having stomata tapering counter to flow direction |
PCT/AU2011/000664 WO2011156841A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for pathogen detection with dialysis, thermal lysis, nucleic acid amplification and prehybridization filtering |
PCT/AU2011/000666 WO2011156843A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for genetic analysis which performs nucleic acid amplification after sample preparation in a dialysis section |
PCT/AU2011/000667 WO2011156844A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for genetic analysis which performs nucleic acid amplification before removing non-nucleic acid constituents in a dialysis section |
PCT/AU2011/000672 WO2011156849A1 (en) | 2010-06-17 | 2011-06-01 | Test module with microfluidic device having loc and dialysis device for separating pathogens from other constituents in a biological sample |
PCT/AU2011/000663 WO2011156840A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for pathogen detection and genetic analysis with chemical lysis, incubation and tandem nucleic acid amplification |
PCT/AU2011/000658 WO2011156835A1 (en) | 2010-06-17 | 2011-06-01 | Test module incorporating spectrometer |
PCT/AU2011/000660 WO2011156837A1 (en) | 2010-06-17 | 2011-06-01 | Loc device for pathogen detection with dialysis, thermal lysis, nucleic acid amplification and prehybridization filtering |
PCT/AU2011/000679 WO2011156856A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic thermal bend actuated surface tension valve |
PCT/AU2011/000676 WO2011156853A1 (en) | 2010-06-17 | 2011-06-01 | Microfluidic test module with flexible membrane for internal microenvironment pressure-relief |
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102649931A (en) * | 2012-05-28 | 2012-08-29 | 上海理工大学 | Preparation method for microarray biochip |
WO2016062788A1 (en) | 2014-10-24 | 2016-04-28 | Ait Austrian Institute Of Technology Gmbh | Microfluidic chip for biological analysis |
Families Citing this family (263)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7790028B1 (en) * | 2005-09-28 | 2010-09-07 | The Charles Stark Draper Laboratory, Inc. | Systems, methods, and devices relating to a cellularized nephron unit |
CN101479605A (en) | 2006-04-21 | 2009-07-08 | 纳诺拜希姆公司 | Single-molecule platform for drug discovery: methods and apparatuses for drug discovery, including discovery of anticancer and antiviralagents |
US11001881B2 (en) | 2006-08-24 | 2021-05-11 | California Institute Of Technology | Methods for detecting analytes |
US8048626B2 (en) | 2006-07-28 | 2011-11-01 | California Institute Of Technology | Multiplex Q-PCR arrays |
US11525156B2 (en) | 2006-07-28 | 2022-12-13 | California Institute Of Technology | Multiplex Q-PCR arrays |
US11560588B2 (en) | 2006-08-24 | 2023-01-24 | California Institute Of Technology | Multiplex Q-PCR arrays |
US8498695B2 (en) | 2006-12-22 | 2013-07-30 | Novadaq Technologies Inc. | Imaging system with a single color image sensor for simultaneous fluorescence and color video endoscopy |
US7993292B2 (en) * | 2007-05-22 | 2011-08-09 | Bellacure, Inc. | Orthotic apparatus and method of operation |
US9598737B2 (en) * | 2012-05-09 | 2017-03-21 | Longhorn Vaccines And Diagnostics, Llc | Next generation genomic sequencing methods |
US8406860B2 (en) | 2008-01-25 | 2013-03-26 | Novadaq Technologies Inc. | Method for evaluating blush in myocardial tissue |
US8707452B1 (en) * | 2008-04-14 | 2014-04-22 | Avaya Inc. | Secure data management device |
EP2927350A1 (en) * | 2014-04-02 | 2015-10-07 | Beatrice Sala | Electrochemical cell for the electrolysis of water in steam or liquid form, manufacturing method and uses |
US8304185B2 (en) * | 2009-07-17 | 2012-11-06 | Canon U.S. Life Sciences, Inc. | Methods and systems for DNA isolation on a microfluidic device |
DE112010002222B4 (en) | 2009-06-04 | 2024-01-25 | Leidos Innovations Technology, Inc. (n.d.Ges.d. Staates Delaware) | Multi-sample microfluidic chip for DNA analysis |
JP5879267B2 (en) * | 2009-10-29 | 2016-03-08 | ザ・チャールズ・スターク・ドレイパ・ラボラトリー・インコーポレイテッド | Microfluidic device for hemodialysis |
US20130040374A1 (en) * | 2010-04-28 | 2013-02-14 | Panasonic Corporation | Chemical sensor |
US20110312612A1 (en) * | 2010-06-17 | 2011-12-22 | Geneasys Pty Ltd | Loc device for electrochemiluminescent detection of target sequences with probes between a working electrode and a photosensor |
US10161928B2 (en) | 2010-07-26 | 2018-12-25 | Wellmetris, Llc | Wellness panel |
US20120045786A1 (en) * | 2010-08-19 | 2012-02-23 | Stith Curtis W | Opto-fluidic microscope diagnostic system |
US20120044339A1 (en) * | 2010-08-19 | 2012-02-23 | Stith Curtis W | Opto-fluidic microscope system with evaluation chambers |
CA2809581C (en) * | 2010-08-27 | 2016-11-15 | The Board Of Trustees Of The Leland Stanford Junior University | Microscopy imaging device with advanced imaging properties |
WO2012040050A1 (en) | 2010-09-23 | 2012-03-29 | Bayer Healthcare Llc | System and method for determining ambient temperatures for a fluid analyte system |
US8961764B2 (en) | 2010-10-15 | 2015-02-24 | Lockheed Martin Corporation | Micro fluidic optic design |
EP2648609B1 (en) | 2010-12-09 | 2018-05-30 | Zoll Medical Corporation | Electrode with redundant impedance reduction |
FR2968532B1 (en) * | 2010-12-14 | 2013-04-26 | Commissariat Energie Atomique | DEVICE AND METHOD FOR DETERMINING AN EXCRETION RATE OF A BODILY FLUID BY AN INDIVIDUAL OR ANIMAL |
CN102564576B (en) * | 2010-12-17 | 2013-11-06 | 鸿富锦精密工业(深圳)有限公司 | Light intensity testing device |
DK2665557T3 (en) * | 2011-01-21 | 2020-04-06 | Biodot Inc | Piezoelectric dispenser with a longitudinal transducer and interchangeable capillary tube |
US8794050B2 (en) | 2011-01-27 | 2014-08-05 | Nanoscopia (Cayman), Inc. | Fluid sample analysis systems |
US9469871B2 (en) | 2011-04-14 | 2016-10-18 | Corporos Inc. | Methods and apparatus for point-of-care nucleic acid amplification and detection |
TWI441940B (en) * | 2011-06-09 | 2014-06-21 | Shih Hua Technology Ltd | Method for making pattern conductive element |
JP6041872B2 (en) | 2011-06-15 | 2016-12-14 | ザ チャールズ スターク ドレイパー ラボラトリー インク | Bioartificial kidney |
ITTO20110567A1 (en) * | 2011-06-28 | 2012-12-29 | St Microelectronics Srl | CARTRIDGE FOR BIOCHEMICAL ANALYSIS, BIOCHEMICAL ANALYSIS SYSTEM AND METHOD TO PERFORM A BIOCHEMICAL PROCESS |
TW201301141A (en) * | 2011-06-29 | 2013-01-01 | Walton Advanced Eng Inc | Storage device having graphs and recognition system thereof |
US8717556B2 (en) | 2011-07-27 | 2014-05-06 | Aptina Imaging Corporation | Microfluidic systems with chemical pumps |
CN103733047B (en) * | 2011-08-11 | 2016-03-30 | 奥林巴斯株式会社 | The detection method of intended particle |
EP2752655A4 (en) | 2011-08-30 | 2015-06-17 | Olympus Corp | Method for detecting target particles |
US8988684B1 (en) | 2011-09-08 | 2015-03-24 | Lawrence Livermore National Security, Llc | System and method for measuring fluorescence of a sample |
US9372135B1 (en) | 2011-09-08 | 2016-06-21 | Lawrence Livermore National Security, Llc | Fluidics platform and method for sample preparation |
US9968258B2 (en) * | 2011-09-12 | 2018-05-15 | Tufts University | Imaging fluorescence or luminescence lifetime |
USD753311S1 (en) | 2011-10-12 | 2016-04-05 | Alere Switzerland Gmbh | Isothermal nucleic acid amplification meter |
GB201119032D0 (en) | 2011-11-03 | 2011-12-14 | Isis Innovation | Multisomes: encapsulated droplet networks |
US9689029B2 (en) | 2011-12-02 | 2017-06-27 | Caliper Life Sciences, Inc. | Systems and methods for sampling of amplification products |
US9725761B2 (en) | 2011-12-28 | 2017-08-08 | Ricardo Mancebo | Reagents and methods for autoligation chain reaction |
WO2013112877A1 (en) | 2012-01-25 | 2013-08-01 | Tasso, Inc. | Handheld device for drawing, collecting, and analyzing bodily fluid |
WO2013125124A1 (en) | 2012-02-22 | 2013-08-29 | オリンパス株式会社 | Method for detecting target particles |
TWI484154B (en) * | 2012-02-24 | 2015-05-11 | Optical detecting apparatus and operating method thereof | |
WO2013140890A1 (en) | 2012-03-21 | 2013-09-26 | オリンパス株式会社 | Method for detecting target nucleic acid molecule |
US9150907B2 (en) | 2012-04-27 | 2015-10-06 | General Electric Company | Microfluidic flow cell assemblies and method of use |
US9354159B2 (en) | 2012-05-02 | 2016-05-31 | Nanoscopia (Cayman), Inc. | Opto-fluidic system with coated fluid channels |
US9258536B2 (en) * | 2012-05-03 | 2016-02-09 | Semiconductor Components Industries, Llc | Imaging systems with plasmonic color filters |
US9213043B2 (en) | 2012-05-15 | 2015-12-15 | Wellstat Diagnostics, Llc | Clinical diagnostic system including instrument and cartridge |
US9081001B2 (en) | 2012-05-15 | 2015-07-14 | Wellstat Diagnostics, Llc | Diagnostic systems and instruments |
US9625465B2 (en) | 2012-05-15 | 2017-04-18 | Defined Diagnostics, Llc | Clinical diagnostic systems |
KR20130136623A (en) * | 2012-06-05 | 2013-12-13 | 인제대학교 산학협력단 | Apparatus for detecting liquid electric conductivity |
CA2914778A1 (en) | 2012-06-21 | 2013-12-27 | Novadaq Technologies Inc. | Quantification and analysis of angiography and perfusion |
AU2013293078B2 (en) * | 2012-07-23 | 2017-09-07 | Tasso, Inc. | Methods, systems, and devices relating to open microfluidic channels |
US20140200167A1 (en) | 2012-08-01 | 2014-07-17 | Nanomdx, Inc. | Functionally integrated device for multiplex genetic identification |
US9310300B2 (en) * | 2012-08-03 | 2016-04-12 | Ingeneron Incorporated | Compact portable apparatus for optical assay |
US20140046600A1 (en) * | 2012-08-07 | 2014-02-13 | Netanel Avner | Sim card based medical testing and data transmission system |
CA2879729A1 (en) * | 2012-08-07 | 2014-02-13 | California Institute Of Technology | Ultrafast thermal cycler |
US9580679B2 (en) * | 2012-09-21 | 2017-02-28 | California Institute Of Technology | Methods and devices for sample lysis |
US9518914B2 (en) * | 2012-09-24 | 2016-12-13 | Brigham And Women's Hospital, Inc. | Portal and method for management of dialysis therapy |
DE102012109317A1 (en) * | 2012-10-01 | 2014-04-03 | Astrium Gmbh | Device for carrying out a biochemical analysis, in particular in space |
US9804149B2 (en) * | 2012-10-10 | 2017-10-31 | Bio-Rad Laboratories, Inc. | Patient-based results display |
GB201219201D0 (en) | 2012-10-25 | 2012-12-12 | Isis Innovation | Hydrogel network |
EP3722758B1 (en) * | 2012-11-13 | 2023-11-01 | Viavi Solutions Inc. | Portable spectrometer |
US9885655B2 (en) | 2012-11-13 | 2018-02-06 | Viavi Solutions Inc. | Spectrometer with a relay lightpipe |
JPWO2014077029A1 (en) * | 2012-11-13 | 2017-01-05 | 株式会社村田製作所 | Droplet quantification method and measuring apparatus |
CN103852105A (en) * | 2012-12-04 | 2014-06-11 | 昆山平成电子科技有限公司 | Multi-functional tester |
CN105188934B (en) * | 2012-12-07 | 2018-12-04 | 牛津大学创新有限公司 | The drop component of 3D printing |
US9239328B2 (en) | 2012-12-17 | 2016-01-19 | Taiwan Semiconductor Manufacturing Company, Ltd. | Systems and methods for an integrated bio-entity manipulation and processing semiconductor device |
US9495332B2 (en) * | 2012-12-21 | 2016-11-15 | International Business Machines Corporation | Detection and repositioning of pop-up dialogs |
WO2014105946A1 (en) * | 2012-12-27 | 2014-07-03 | Medi-Physics, Inc. | Dual-filter dual-integrity test assembly |
GB201301178D0 (en) | 2013-01-23 | 2013-03-06 | Dynamic Biosensors Gmbh | Method for sequencing a template nucleic acid immobilized on a substrate |
US9999393B2 (en) | 2013-01-29 | 2018-06-19 | Zoll Medical Corporation | Delivery of electrode gel using CPR puck |
US8880196B2 (en) | 2013-03-04 | 2014-11-04 | Zoll Medical Corporation | Flexible therapy electrode |
SG11201507586PA (en) | 2013-03-12 | 2015-10-29 | Univ Michigan | Microfluidic device for immunoblotting |
WO2014153216A2 (en) * | 2013-03-14 | 2014-09-25 | Alfred E. Mann Foundation For Scientific Research | Microfluidic flow rate sensor |
JP6165961B2 (en) | 2013-03-15 | 2017-07-19 | アボット・ラボラトリーズAbbott Laboratories | Diagnostic analyzer with pre-process carousel and associated method |
CN105142789A (en) * | 2013-03-15 | 2015-12-09 | 纳诺拜希姆公司 | Systems and methods for mobile device analysis of nucleic acids and proteins |
US10933417B2 (en) | 2013-03-15 | 2021-03-02 | Nanobiosym, Inc. | Systems and methods for mobile device analysis of nucleic acids and proteins |
GB201304797D0 (en) * | 2013-03-15 | 2013-05-01 | Diagnostics For The Real World Ltd | Apparatus and method for automated sample preparation and adaptor for use in the apparatus |
CN107831324B (en) | 2013-03-15 | 2021-11-19 | 雅培制药有限公司 | Automated diagnostic analyzer with rear accessible track system and related methods |
CN109358202B (en) | 2013-03-15 | 2023-04-07 | 雅培制药有限公司 | Automated diagnostic analyzer with vertically arranged carousel and related methods |
KR101483493B1 (en) * | 2013-03-22 | 2015-01-19 | 나노바이오시스 주식회사 | PCR device for detecting food-borne bacteria, and and method for detecting food-borne bacteria using the same |
US20140323819A1 (en) * | 2013-04-29 | 2014-10-30 | Elwha LLC, a limited liability company of the State of Delaware | Multi-parameter test units for initial indication of medical symptoms |
WO2014194042A2 (en) * | 2013-05-29 | 2014-12-04 | Gnubio, Inc. | Low cost optical high speed discrete measurement system |
GB2516669B (en) * | 2013-07-29 | 2015-09-09 | Atlas Genetics Ltd | A method for processing a liquid sample in a fluidic cartridge |
US9347962B2 (en) | 2013-08-05 | 2016-05-24 | Nanoscopia (Cayman), Inc. | Handheld diagnostic system with chip-scale microscope and automated image capture mechanism |
US10203284B2 (en) | 2013-08-26 | 2019-02-12 | Diagenetix, Inc. | Hardware and mobile software for operation of portable instruments for nucleic acid amplification |
JP2016534839A (en) * | 2013-09-16 | 2016-11-10 | ジョンソン・アンド・ジョンソン・イノベーション・エルエルシーJohnson & Johnson Innovation LLC | Microfluidic organ assist device incorporating boundary layer separation |
ITMI20131541A1 (en) * | 2013-09-19 | 2015-03-20 | Eugenio Iannone | DIAGNOSIS DEVICE, PARTICULARLY OF THE LAB-ON-CHIP TYPE. |
US20160215254A1 (en) | 2013-09-27 | 2016-07-28 | Deirdre Meldrum | System and method for laser lysis |
US9604214B2 (en) * | 2013-10-01 | 2017-03-28 | Owl biomedical, Inc. | Cell sorting system using microfabricated components |
DK3066219T3 (en) * | 2013-11-08 | 2019-03-11 | Ionis Pharmaceuticals Inc | METHODS FOR DETECTING OIGONUCLEOTIDES |
US9059337B1 (en) * | 2013-12-24 | 2015-06-16 | Christie Digital Systems Usa, Inc. | Method, system and apparatus for dynamically monitoring and calibrating display tiles |
US20180135108A1 (en) * | 2014-01-20 | 2018-05-17 | Board Of Trustees Of Michigan State University | Method for detecting bacterial and fungal pathogens |
US10260111B1 (en) | 2014-01-20 | 2019-04-16 | Brett Eric Etchebarne | Method of detecting sepsis-related microorganisms and detecting antibiotic-resistant sepsis-related microorganisms in a fluid sample |
WO2015123603A1 (en) * | 2014-02-14 | 2015-08-20 | Beth Israel Deaconess Medical Center, Inc. | Electrical impedance myography |
EP3110428A4 (en) * | 2014-02-28 | 2017-11-22 | Bdbc Sciences Corp. | A system for tissue manipulation |
EP3822635A1 (en) | 2014-03-07 | 2021-05-19 | The Regents Of The University Of California | Devices for integrating analyte extraction, concentration and detection |
WO2015138343A1 (en) | 2014-03-10 | 2015-09-17 | Click Diagnostics, Inc. | Cartridge-based thermocycler |
WO2015138648A1 (en) | 2014-03-11 | 2015-09-17 | Illumina, Inc. | Disposable, integrated microfluidic cartridge and methods of making and using same |
MX2016011625A (en) * | 2014-03-12 | 2016-12-09 | Basf Se | Improved catalyzed soot filter. |
US8820538B1 (en) | 2014-03-17 | 2014-09-02 | Namocell LLC | Method and apparatus for particle sorting |
US10318878B2 (en) | 2014-03-19 | 2019-06-11 | Numenta, Inc. | Temporal processing scheme and sensorimotor information processing |
GB2524730A (en) * | 2014-03-30 | 2015-10-07 | Robert Faulkner | Predicting application user behaviour and interests in real-time using predictive analytics as a service |
CN106460233A (en) * | 2014-04-14 | 2017-02-22 | 思研(Sri)国际顾问与咨询公司 | Portable nucleic acid analysis systemand high-performance microfluidic electroactive polymer actuators |
JP2017520776A (en) * | 2014-04-25 | 2017-07-27 | ヒューレット−パッカード デベロップメント カンパニー エル.ピー.Hewlett‐Packard Development Company, L.P. | Diagnostic cassette |
CN104668002B (en) * | 2014-05-22 | 2016-03-16 | Imec非营利协会 | Compact fluid analysis device and manufacture method thereof |
CA2991918A1 (en) * | 2014-07-11 | 2016-01-14 | Advanced Theranostics Inc. | Point of care polymerase chain reaction device for disease detection |
JP2016023994A (en) * | 2014-07-17 | 2016-02-08 | セイコーエプソン株式会社 | Electronic component conveyance device and electronic component inspection device |
EP3769682B1 (en) | 2014-08-01 | 2024-01-03 | Tasso, Inc. | Systems for gravity-enhanced microfluidic collection, handling and transferring of fluids |
US9921182B2 (en) | 2014-10-06 | 2018-03-20 | ALVEO Technologies Inc. | System and method for detection of mercury |
US10352899B2 (en) | 2014-10-06 | 2019-07-16 | ALVEO Technologies Inc. | System and method for detection of silver |
US10196678B2 (en) | 2014-10-06 | 2019-02-05 | ALVEO Technologies Inc. | System and method for detection of nucleic acids |
US10627358B2 (en) | 2014-10-06 | 2020-04-21 | Alveo Technologies, Inc. | Method for detection of analytes |
US9506908B2 (en) | 2014-10-06 | 2016-11-29 | Alveo Technologies, Inc. | System for detection of analytes |
US10022718B2 (en) * | 2014-10-24 | 2018-07-17 | Arizona Board Of Regents On Behalf Of Arizona State University | Microfluidic device and array disk |
CN104374903B (en) * | 2014-11-08 | 2016-07-06 | 东莞博识生物科技有限公司 | A kind of in-vitro diagnosis test card |
CN114958990A (en) | 2014-12-31 | 2022-08-30 | 维斯比医学公司 | Method for detecting target nucleic acid using molecular diagnostic test device |
RU2016151351A (en) * | 2015-01-28 | 2019-02-28 | КАБУСИКИ КАЙСЯ ДиЭнЭйФОРМ | ANALYSIS DEVICE, ANALYSIS CHIP, ANALYSIS KIT AND METHOD OF ANALYSIS WITH THEIR APPLICATION |
KR102302835B1 (en) | 2015-01-30 | 2021-09-15 | 휴렛-팩커드 디벨롭먼트 컴퍼니, 엘.피. | diagnostic chip |
DE102015001998B3 (en) * | 2015-02-20 | 2016-02-04 | Friz Biochem Gesellschaft Für Bioanalytik Mbh | Microfluidic cartridge for the detection of biomolecules |
US10279352B2 (en) | 2015-03-18 | 2019-05-07 | Optolane Technologies Inc. | PCR module, PCR system having the same, and method of inspecting using the same |
MX2017012095A (en) * | 2015-03-23 | 2018-09-21 | Wellmetris Llc | Smartphone enabled urinalysis device, software, and test platform. |
US9708647B2 (en) | 2015-03-23 | 2017-07-18 | Insilixa, Inc. | Multiplexed analysis of nucleic acid hybridization thermodynamics using integrated arrays |
US10495507B2 (en) * | 2015-04-30 | 2019-12-03 | Hewlett-Packard Development Company, L.P. | Drop ejection based flow sensor calibration |
CN107430018B (en) * | 2015-04-30 | 2022-03-18 | 惠普发展公司,有限责任合伙企业 | Microfluidic flow sensor |
WO2016188738A1 (en) * | 2015-05-26 | 2016-12-01 | F. Hoffmann-La Roche Ag | Point of care testing poct system |
CN104833747B (en) * | 2015-05-06 | 2016-08-24 | 华东理工大学 | A kind of preparative hplc UV-detector using deep ultraviolet LED light source |
JP6933583B2 (en) | 2015-05-20 | 2021-09-08 | ユニバーシティ オブ メリーランド, カレッジ パーク | Generation and capture of water droplets in microfluidic chips with continuous gas phase |
AU2016318103B2 (en) | 2015-09-04 | 2022-11-17 | The Regents Of The University Of California | Methods and devices for analyte collection, extraction, concentration, and detection for clinical applications |
US9499861B1 (en) | 2015-09-10 | 2016-11-22 | Insilixa, Inc. | Methods and systems for multiplex quantitative nucleic acid amplification |
US9735305B2 (en) | 2015-09-21 | 2017-08-15 | International Business Machines Corporation | Monolithically integrated fluorescence on-chip sensor |
WO2017059094A2 (en) * | 2015-09-29 | 2017-04-06 | Adi Mashiach | System and method for detection of disease in bodily fluids |
CN113870961A (en) * | 2015-10-01 | 2021-12-31 | 基因动力公司 | Method, tool and system for providing analysis for biological sample test results to a user |
AU2016339957C1 (en) * | 2015-10-16 | 2021-08-26 | Opko Diagnostics, Llc | Articles and methods for preparing a surface for obtaining a patient sample |
AU2016351730B2 (en) | 2015-11-13 | 2019-07-11 | Novadaq Technologies Inc. | Systems and methods for illumination and imaging of a target |
WO2017087703A1 (en) * | 2015-11-17 | 2017-05-26 | Nanoscopia (Cayman), Inc. | Sample processing and smearing apparatus and methods |
CN108291875B (en) * | 2015-11-26 | 2021-05-07 | 富士胶片株式会社 | Solution adhesion device and solution adhesion method |
AU2016377659B9 (en) | 2015-12-21 | 2021-11-11 | Tasso, Inc. | Devices, systems and methods for actuation and retraction in fluid collection |
EP3394293B1 (en) * | 2015-12-22 | 2021-05-26 | Canon U.S. Life Sciences, Inc. | Sample-to-answer system for microorganism detection featuring target enrichment, amplification and detection |
WO2017117666A1 (en) | 2016-01-08 | 2017-07-13 | Advanced Theranostics Inc. | Fully integrated, stand-alone, point-of-care device to detect target nucleic acids |
JP1565699S (en) * | 2016-01-12 | 2016-12-19 | ||
US10436773B2 (en) * | 2016-01-18 | 2019-10-08 | Jana Care, Inc. | Mobile device based multi-analyte testing analyzer for use in medical diagnostic monitoring and screening |
WO2017127929A1 (en) | 2016-01-26 | 2017-08-03 | Novadaq Technologies Inc. | Configurable platform |
US9643181B1 (en) | 2016-02-08 | 2017-05-09 | International Business Machines Corporation | Integrated microfluidics system |
JP6274233B2 (en) * | 2016-02-23 | 2018-02-07 | 住友ベークライト株式会社 | Clip cartridge system |
WO2017155858A1 (en) | 2016-03-07 | 2017-09-14 | Insilixa, Inc. | Nucleic acid sequence identification using solid-phase cyclic single base extension |
CN105675371B (en) * | 2016-03-29 | 2018-09-25 | 广东江门生物技术开发中心有限公司 | A kind of Multifunctional inspection sample extraction separator |
WO2017181069A1 (en) | 2016-04-15 | 2017-10-19 | University Of Maryland, College Park | Integrated thermoplastic chip for rapid pcr and hrma |
WO2017185067A1 (en) * | 2016-04-22 | 2017-10-26 | Click Diagnostics, Inc. | Printed circuit board heater for an amplification module |
WO2017197040A1 (en) | 2016-05-11 | 2017-11-16 | Click Diagnostics, Inc. | Devices and methods for nucleic acid extraction |
CN107400628B (en) * | 2016-05-19 | 2021-03-02 | 深圳市华因康高通量生物技术研究院 | Sequencing reaction cell, sequencing reaction clamp and sequencing reaction equipment |
CN106018389A (en) * | 2016-05-20 | 2016-10-12 | 华南师范大学 | Handheld POCT (Point of Care Testing) bipolar electrode-electrochemical light emitting device and application thereof |
KR102592388B1 (en) | 2016-06-09 | 2023-10-20 | 더 리전트 오브 더 유니버시티 오브 캘리포니아 | A single platform for biomarker enrichment and signal amplification for use in paper-based immunoassays, and for extraction, concentration and amplification of DNA. |
EP3469420A4 (en) | 2016-06-14 | 2020-02-12 | Novadaq Technologies ULC | Methods and systems for adaptive imaging for low light signal enhancement in medical visualization |
JP6729027B2 (en) * | 2016-06-15 | 2020-07-22 | ウシオ電機株式会社 | Micro channel chip |
MX2018015889A (en) * | 2016-06-29 | 2019-05-27 | Click Diagnostics Inc | Devices and methods for the detection of molecules using a flow cell. |
CN106026006B (en) * | 2016-06-30 | 2019-07-16 | 成绎半导体技术(上海)有限公司 | A kind of USB Type-C interface female intelligent measurement and protection circuit |
CA3029682A1 (en) * | 2016-06-30 | 2018-01-04 | Click Diagnostics, Inc. | Devices and methods for nucleic acid extraction |
JP6632487B2 (en) * | 2016-07-13 | 2020-01-22 | キヤノン株式会社 | Continuum robot, method of correcting kinematics, and control method of continuum robot |
WO2018017134A1 (en) * | 2016-07-22 | 2018-01-25 | Hewlett-Packard Development Company, L.P. | Substrate assembly and related methods |
US10889854B2 (en) * | 2016-08-08 | 2021-01-12 | Universiti Brunei Darussalam | System and method for immobilization free electrochemiluminescence DNA detection using a luminophore dye for multi-species detection |
CN106323353B (en) * | 2016-08-12 | 2019-02-12 | Oppo广东移动通信有限公司 | A kind of calibration method of proximity sensor, device and terminal |
WO2018039139A1 (en) | 2016-08-22 | 2018-03-01 | The Regents Of The University Of California | Hydrogel platform for aqueous two-phase concentration of a target to enhance its detection |
CN109996888A (en) | 2016-09-23 | 2019-07-09 | 阿尔韦奥科技公司 | For testing and analyzing the method and composition of object |
WO2018071541A1 (en) * | 2016-10-11 | 2018-04-19 | The Regents Of The University Of California | Integrated molecular diagnostics system (imdx) and method for dengue fever |
CA3041043A1 (en) * | 2016-10-24 | 2018-05-03 | The Trustees Of The University Of Pennsylvania | Ultra-high throughput detection of fluorescent droplets using time domain encoded optofluidics |
JP6671665B2 (en) * | 2016-10-27 | 2020-03-25 | シャープ株式会社 | Fluorescence inspection system, dielectrophoresis device and molecular inspection method |
WO2018094115A1 (en) | 2016-11-16 | 2018-05-24 | Catalog Technologies, Inc. | Systems for nucleic acid-based data storage |
US10650312B2 (en) | 2016-11-16 | 2020-05-12 | Catalog Technologies, Inc. | Nucleic acid-based data storage |
PL235210B1 (en) * | 2016-12-21 | 2020-06-15 | Genomtec Spolka Akcyjna | Method for detection of genetic material in a biological specimen the device for the execution of this method |
GB201704754D0 (en) | 2017-01-05 | 2017-05-10 | Illumina Inc | Kinetic exclusion amplification of nucleic acid libraries |
WO2018144906A1 (en) * | 2017-02-02 | 2018-08-09 | University Of Maryland, College Park | Trap arrays for robust microfluidic sample digitization |
EP3576881A4 (en) * | 2017-02-06 | 2019-12-25 | EFA - Engineering For All Ltd. | Portable digital diagnostic device |
JP6931705B2 (en) * | 2017-02-10 | 2021-09-08 | ノバダック テクノロジーズ ユーエルシー | Open Field Handheld Fluorescence Imaging Systems and Methods |
US11209427B2 (en) | 2017-03-27 | 2021-12-28 | The Regents Of The University Of California | Semi-quantitative lateral-flow immunoassay for the detection of CSF leaks |
CN106895908B (en) * | 2017-03-29 | 2018-10-16 | 中国汽车技术研究中心 | Check device during a kind of high stability laser positioning luminosity probe |
USD849265S1 (en) * | 2017-04-21 | 2019-05-21 | Precision Nanosystems Inc | Microfluidic chip |
US11441701B2 (en) | 2017-07-14 | 2022-09-13 | Hewlett-Packard Development Company, L.P. | Microfluidic valve |
WO2019022753A1 (en) | 2017-07-28 | 2019-01-31 | Hewlett-Packard Development Company, L.P. | Ionic species interrogation and sensing |
JP2019024453A (en) * | 2017-08-02 | 2019-02-21 | 株式会社リコー | Rna concentration quantitative method, rna concentration quantitative device, and rna concentration quantitative apparatus |
CN208016344U (en) * | 2017-08-30 | 2018-10-30 | 苏州宝时得电动工具有限公司 | Automatic running device |
US11327090B2 (en) | 2017-09-27 | 2022-05-10 | Hewlett-Packard Development Company, L.P. | Reuse of dispensers during alignment procedures |
US11402400B2 (en) | 2017-10-13 | 2022-08-02 | Hewlett-Packard Development Company, L.P. | Partition liquid into samples |
CN111655866A (en) | 2017-11-09 | 2020-09-11 | 维斯比医学公司 | Portable molecular diagnostic device and method for detecting target virus |
US11383236B2 (en) * | 2017-11-10 | 2022-07-12 | Christopher Walker | Polymerase chain reaction using a microfluidic chip fabricated with printed circuit board techniques |
CN107967380B (en) * | 2017-11-15 | 2021-09-07 | 晶晨半导体(上海)股份有限公司 | Layout design method of printed circuit board |
US10854251B2 (en) * | 2017-12-15 | 2020-12-01 | Google Llc | Physical identifiers for authenticating an identity of a semiconductor component |
CN108221844B (en) * | 2017-12-31 | 2023-01-10 | 浙江大学 | Dynamic response test device for near-sea foundation pit under effect of simulated tidal load |
US20200360929A1 (en) * | 2018-01-29 | 2020-11-19 | The Texas A&M University System | Integrated Modular On-Chip Droplet Microfluidic Screening Platform |
CN108195728A (en) * | 2018-02-01 | 2018-06-22 | 山东诺方电子科技有限公司 | A kind of control system and its control method based on multinuclear particulate matter sensors technology |
CA3092684A1 (en) * | 2018-03-02 | 2019-09-06 | Teleflex Medical Incorporated | Infection detection systems and methods |
EP3766077A4 (en) | 2018-03-16 | 2021-12-08 | Catalog Technologies, Inc. | Chemical methods for nucleic acid-based data storage |
WO2019222561A1 (en) | 2018-05-16 | 2019-11-21 | Catalog Technologies, Inc. | Compositions and methods for nucleic acid-based data storage |
EP3758844B1 (en) * | 2018-06-11 | 2023-03-29 | Hewlett-Packard Development Company, L.P. | Microfluidic valves |
US10307755B1 (en) * | 2018-07-19 | 2019-06-04 | Bioceryx Inc. | Apparatuses and methods for sample-specific self-configuration |
US10883914B2 (en) | 2018-08-07 | 2021-01-05 | Blaire Biomedical, LLC | Flow cytometry systems including an optical analysis box for interfacing with an imaging device |
CN110819522B (en) * | 2018-08-13 | 2023-09-22 | 上海新微技术研发中心有限公司 | Digital PCR system and digital PCR liquid drop forming method |
JP2021536015A (en) * | 2018-09-14 | 2021-12-23 | ウィリアム マーシュ ライス ユニバーシティWilliam Marsh Rice University | Equipment and Methods for Multiple Amplification and Detection of DNA Using Convection Heating and Unlabeled Microarrays |
JP7460607B2 (en) | 2018-09-14 | 2024-04-02 | タッソ インコーポレイテッド | Body fluid collection devices and related methods |
US11486814B2 (en) * | 2018-10-01 | 2022-11-01 | Hewlett-Packard Development Company, L.P. | Particle sorting using microfluidic ejectors |
CN112752964A (en) * | 2018-10-01 | 2021-05-04 | 惠普发展公司,有限责任合伙企业 | Bulk particle sorting |
US20210239958A1 (en) * | 2018-10-01 | 2021-08-05 | Hewlett-Packard Development Company, L.P. | Microscopy systems |
JP6795019B2 (en) * | 2018-10-04 | 2020-12-02 | カシオ計算機株式会社 | Case and watch |
US11946902B2 (en) * | 2018-10-11 | 2024-04-02 | Hewlett-Packard Development Company, L.P. | Dielectrophoresis separator cross-over frequency measurement systems |
US10760936B2 (en) * | 2018-11-02 | 2020-09-01 | Nanya Technology Corporation | Semiconductor device and method of sensing a change in a level of a liquid therein |
FR3088534A1 (en) * | 2018-11-16 | 2020-05-22 | Commissariat A L'energie Atomique Et Aux Energies Alternatives | DEVICE FOR PREPARING A CALIBRATED VOLUME OF BLOOD PLASMA |
US20210354126A1 (en) * | 2018-12-07 | 2021-11-18 | Ultima Genomics, Inc. | Implementing barriers for controlled environments during sample processing and detection |
WO2020122922A1 (en) * | 2018-12-13 | 2020-06-18 | Hewlett-Packard Development Company, L.P. | Multiplex nucleic acid detection |
MX2021007297A (en) * | 2018-12-20 | 2021-09-08 | Alveo Tech Inc | Handheld impedance-based diagnostic test system for detecting analytes. |
WO2020149926A1 (en) * | 2019-01-17 | 2020-07-23 | Siemens Healthcare Diagnostics Inc. | Flow cell using peltier module as prime mover for polymerase chain reaction |
EP3880841B1 (en) | 2019-02-22 | 2023-05-31 | Hewlett-Packard Development Company, L.P. | Nucleic acid detection |
EP3937780A4 (en) | 2019-03-14 | 2022-12-07 | InSilixa, Inc. | Methods and systems for time-gated fluorescent-based detection |
CN110180811A (en) * | 2019-05-07 | 2019-08-30 | 王海山 | A kind of chip detecting equipment that the clarity with dehumidification function is high |
EP3966823A1 (en) | 2019-05-09 | 2022-03-16 | Catalog Technologies, Inc. | Data structures and operations for searching, computing, and indexing in dna-based data storage |
WO2020246963A1 (en) * | 2019-06-04 | 2020-12-10 | Hewlett-Packard Development Company, L.P. | Integrated microfluidic ejector chips |
US20220347675A1 (en) * | 2019-07-24 | 2022-11-03 | Hewlett-Packard Development Company, L.P. | Device with microfluidic channels |
EP3769840A1 (en) * | 2019-07-26 | 2021-01-27 | LEX Diagnostics Ltd | Systems and modules for nucleic acid amplification testing |
GB201911386D0 (en) | 2019-08-09 | 2019-09-25 | Stratec Se | Calibration tool for planar chip applications |
CN110333272A (en) * | 2019-08-21 | 2019-10-15 | 业成科技(成都)有限公司 | Humidity sensor and its manufacturing method |
WO2021046504A1 (en) * | 2019-09-06 | 2021-03-11 | Teleflex Medical Incorporated | Infection detection systems and methods including a sample processor having integrated sample filter and meter |
WO2021046506A1 (en) * | 2019-09-06 | 2021-03-11 | Teleflex Medical Incorporated | Infection detection systems and methods including intermediate filtering and metering |
AU2020361681A1 (en) * | 2019-10-10 | 2022-05-05 | 1859, Inc. | Methods and systems for microfluidic screening |
KR20220080172A (en) | 2019-10-11 | 2022-06-14 | 카탈로그 테크놀로지스, 인크. | Nucleic Acid Security and Authentication |
USD954573S1 (en) * | 2019-11-06 | 2022-06-14 | Fluxergy, Llc | Test card |
EP4085149A4 (en) | 2020-01-03 | 2024-03-06 | Visby Medical Inc | Devices and methods for antibiotic susceptibility testing |
US11136543B1 (en) | 2020-02-11 | 2021-10-05 | Edward R. Flynn | Magnetic cell incubation device |
US11536732B2 (en) | 2020-03-13 | 2022-12-27 | Jana Care, Inc. | Devices, systems, and methods for measuring biomarkers in biological fluids |
KR20210128632A (en) * | 2020-04-17 | 2021-10-27 | 커넥타젠(주) | Apparatus for Detecting Potable Isothermal Amplification |
TWI749529B (en) * | 2020-04-20 | 2021-12-11 | 關鍵禾芯科技股份有限公司 | Ribonucleic acid test panel and ribonucleic acid test device |
CN111534430B (en) * | 2020-04-28 | 2023-12-29 | 港岫科技(上海)有限公司 | Ribonucleic acid detection panel and ribonucleic acid detection device |
JP2023524117A (en) * | 2020-04-30 | 2023-06-08 | スタブ ヴィーダ - インヴェスティガサォン エ セルヴィソス エム シエンシアス バイオロジカス エルディーエー | Method and portable device for detecting nucleic acid sequences in suspected coronavirus samples |
CA3183416A1 (en) | 2020-05-11 | 2021-11-18 | Catalog Technologies, Inc. | Programs and functions in dna-based data storage |
US20210354127A1 (en) * | 2020-05-13 | 2021-11-18 | Keycore Technology Corp. | Ribonucleic acid test panel and ribonucleic acid test device |
US11654436B2 (en) | 2020-08-11 | 2023-05-23 | Seagate Technology Llc | Microwave heating device for lab on a chip |
CN114317223A (en) * | 2020-09-30 | 2022-04-12 | 富佳生技股份有限公司 | Nucleic acid detecting cassette and nucleic acid detecting apparatus |
JP2022058244A (en) * | 2020-09-30 | 2022-04-11 | 富佳生技股▲ふん▼有限公司 | Nucleic acid detection box and nucleic acid detection device |
CN112255397B (en) * | 2020-10-16 | 2022-06-07 | 吉林大学 | Kit for detecting Listeria monocytogenes, Vibrio parahaemolyticus and Salmonella typhimurium and preparation method thereof |
CN112275335B (en) * | 2020-10-16 | 2022-06-28 | 吉林大学 | Self-suction valve separation type chip, preparation method and detection method of Listeria monocytogenes |
GB2600103B (en) * | 2020-10-19 | 2024-01-10 | Quantumdx Group Ltd | Integrated thermal conditioning and PCR in a molecular POC diagnostic system |
EP3992613A1 (en) * | 2020-10-28 | 2022-05-04 | Koninklijke Philips N.V. | Sputum analysis method and system |
WO2022099087A1 (en) * | 2020-11-05 | 2022-05-12 | President And Fellows Of Harvard College | An airborne pathogen diagnostic platform |
CN112553054A (en) * | 2020-12-10 | 2021-03-26 | 上海艾众生物科技有限公司 | Cell separation apparatus for bioreactor |
TR202021833A2 (en) * | 2020-12-26 | 2021-10-21 | Bilkent Holding A S | DISPOSABLE PATHOGEN DETECTION CHIP AND A RELATED PRODUCTION METHOD |
CN112858085B (en) * | 2021-01-19 | 2021-11-02 | 竹简云(天津)生物科技有限公司 | Food drug solubility detection and analysis device |
WO2022177558A1 (en) * | 2021-02-17 | 2022-08-25 | Hewlett-Packard Development Company, L.P. | Microfluidic nucleic acid amplification |
US11891671B1 (en) * | 2021-03-24 | 2024-02-06 | A9.Com, Inc. | Virus detection system |
KR102423154B1 (en) * | 2021-04-13 | 2022-07-20 | 주식회사 시큐어메드 | Conductive plastic diagnostic device and manufacturing method thereof |
WO2022232056A1 (en) * | 2021-04-26 | 2022-11-03 | Chan Zuckerberg Biohub, Inc. | Testing devices |
CN113340332B (en) * | 2021-05-27 | 2022-07-12 | 西安交通大学 | Photoelectric sensor calibration device and method |
US11630428B2 (en) * | 2021-08-06 | 2023-04-18 | Trimble Inc. | Real-time analysis of vibration samples for operating environment classification and anomaly detection |
KR20230049323A (en) * | 2021-10-06 | 2023-04-13 | 경희대학교 산학협력단 | Diagnostic microfluidic chip, system and IoT-based genetic analysis system including the same |
TWI797820B (en) * | 2021-11-08 | 2023-04-01 | 財團法人工業技術研究院 | Pcr rapid detection device and method thereof |
CN114308162B (en) * | 2021-12-31 | 2023-05-05 | 北京百奥纳芯生物科技有限公司 | Device for assisting combination and fixation of gene chip probe and substrate |
WO2023152599A1 (en) | 2022-02-08 | 2023-08-17 | Universita' Degli Studi Magna Graecia Di Catanzaro | Platform for screening static and dynamic cell culture supports |
GB202204431D0 (en) * | 2022-03-29 | 2022-05-11 | Enzyre Bv | A sensor for testing biomakers in nano litre volumes of plasma based on luminescence |
WO2024006245A1 (en) * | 2022-07-01 | 2024-01-04 | Owl biomedical, Inc. | Illumination and imaging system in tdi-based continuous line scanning microscopy |
GB2621159A (en) * | 2022-08-04 | 2024-02-07 | Wobble Genomics Ltd | Methods of preparing processed nucleic acid samples and detecting nucleic acids and devices therefor |
CN116188510B (en) * | 2023-04-25 | 2023-07-07 | 安徽皖欣环境科技有限公司 | Enterprise emission data acquisition system based on multiple sensors |
CN116854551B (en) * | 2023-06-29 | 2024-03-29 | 武汉大学 | Solid working medium for improving laser micro-propulsion performance and preparation method and application thereof |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1995025116A1 (en) * | 1994-03-16 | 1995-09-21 | California Institute Of Technology | Method and apparatus for performing multiple sequential reactions on a matrix |
WO1998041531A2 (en) * | 1997-03-20 | 1998-09-24 | University Of Washington | Solvent for biopolymer synthesis, solvent microdroplets and methods of use |
Family Cites Families (276)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3283560A (en) * | 1962-12-24 | 1966-11-08 | Du Pont | Differential thermal analysis apparatus |
DE3110879A1 (en) * | 1981-03-20 | 1982-09-30 | Philips Patentverwaltung Gmbh, 2000 Hamburg | ELECTROCHEMOLUMINESCENT CELL |
US4407290A (en) * | 1981-04-01 | 1983-10-04 | Biox Technology, Inc. | Blood constituent measuring device and method |
US5187990A (en) * | 1984-02-16 | 1993-02-23 | Rainin Instrument Co., Inc. | Method for dispensing liquids with a pipette with compensation for air pressure and surface tension |
US5075447A (en) * | 1984-09-17 | 1991-12-24 | Hoffmann-La Roche Inc. | Ruthenium complexes useful as carriers for immunologically active materials |
US4713347A (en) * | 1985-01-14 | 1987-12-15 | Sensor Diagnostics, Inc. | Measurement of ligand/anti-ligand interactions using bulk conductance |
US5038852A (en) * | 1986-02-25 | 1991-08-13 | Cetus Corporation | Apparatus and method for performing automated amplification of nucleic acid sequences and assays using heating and cooling steps |
US4929426A (en) * | 1987-11-02 | 1990-05-29 | Biologix, Inc. | Portable blood chemistry measuring apparatus |
DE4113033A1 (en) * | 1991-04-20 | 1992-10-22 | Fraunhofer Ges Forschung | INTEGRATED CONDUCTIVITY MEASURING DEVICE |
US5726026A (en) * | 1992-05-01 | 1998-03-10 | Trustees Of The University Of Pennsylvania | Mesoscale sample preparation device and systems for determination and processing of analytes |
US5498392A (en) * | 1992-05-01 | 1996-03-12 | Trustees Of The University Of Pennsylvania | Mesoscale polynucleotide amplification device and method |
US5304487A (en) * | 1992-05-01 | 1994-04-19 | Trustees Of The University Of Pennsylvania | Fluid handling in mesoscale analytical devices |
US6315953B1 (en) * | 1993-11-01 | 2001-11-13 | Nanogen, Inc. | Devices for molecular biological analysis and diagnostics including waveguides |
US5965452A (en) * | 1996-07-09 | 1999-10-12 | Nanogen, Inc. | Multiplexed active biologic array |
US6071699A (en) * | 1996-06-07 | 2000-06-06 | California Institute Of Technology | Nucleic acid mediated electron transfer |
US5700695A (en) * | 1994-06-30 | 1997-12-23 | Zia Yassinzadeh | Sample collection and manipulation method |
US5610635A (en) * | 1994-08-09 | 1997-03-11 | Encad, Inc. | Printer ink cartridge with memory storage capacity |
US6635226B1 (en) * | 1994-10-19 | 2003-10-21 | Agilent Technologies, Inc. | Microanalytical device and use thereof for conducting chemical processes |
JPH08122247A (en) * | 1994-10-24 | 1996-05-17 | Hitachi Ltd | Analyzer |
US5527710A (en) * | 1994-12-02 | 1996-06-18 | Igen, Inc. | Rate measurements of biomolecular reactions using electrochemiluminescence |
US6673533B1 (en) * | 1995-03-10 | 2004-01-06 | Meso Scale Technologies, Llc. | Multi-array multi-specific electrochemiluminescence testing |
US6207369B1 (en) * | 1995-03-10 | 2001-03-27 | Meso Scale Technologies, Llc | Multi-array, multi-specific electrochemiluminescence testing |
US20020022261A1 (en) * | 1995-06-29 | 2002-02-21 | Anderson Rolfe C. | Miniaturized genetic analysis systems and methods |
US5856174A (en) * | 1995-06-29 | 1999-01-05 | Affymetrix, Inc. | Integrated nucleic acid diagnostic device |
US5633724A (en) * | 1995-08-29 | 1997-05-27 | Hewlett-Packard Company | Evanescent scanning of biochemical array |
US6130098A (en) * | 1995-09-15 | 2000-10-10 | The Regents Of The University Of Michigan | Moving microdroplets |
US6114122A (en) * | 1996-03-26 | 2000-09-05 | Affymetrix, Inc. | Fluidics station with a mounting system and method of using |
US5707589A (en) * | 1996-04-12 | 1998-01-13 | Merlin Instrument Company | Funnel-shaped sample-vial septum with membrane covered diffusion-barrier section |
US5798502A (en) * | 1996-05-10 | 1998-08-25 | Oak Frequency | Temperature controlled substrate for VLSI construction having minimal parasitic feedback |
FR2750999B1 (en) * | 1996-07-10 | 1998-11-20 | Appligene Oncor | THERMOSTABLE DNA POLYMERASE OF ARCHAEBACTERIA OF THE GENUS PYROCOCCUS SP |
US6136212A (en) * | 1996-08-12 | 2000-10-24 | The Regents Of The University Of Michigan | Polymer-based micromachining for microfluidic devices |
WO1998022625A1 (en) * | 1996-11-20 | 1998-05-28 | The Regents Of The University Of Michigan | Microfabricated isothermal nucleic acid amplification devices and methods |
US6027459A (en) * | 1996-12-06 | 2000-02-22 | Abbott Laboratories | Method and apparatus for obtaining blood for diagnostic tests |
US6529446B1 (en) * | 1996-12-20 | 2003-03-04 | Telaric L.L.C. | Interactive medication container |
US7314711B2 (en) * | 1997-05-23 | 2008-01-01 | Bioveris Corporation | Assays employing electrochemiluminescent labels and electrochemiluminescence quenchers |
US7160687B1 (en) * | 1997-05-29 | 2007-01-09 | Cellomics, Inc. | Miniaturized cell array methods and apparatus for cell-based screening |
AUPO793797A0 (en) * | 1997-07-15 | 1997-08-07 | Silverbrook Research Pty Ltd | A method of manufacture of an image creation apparatus (IJM03) |
US7328975B2 (en) * | 1997-07-15 | 2008-02-12 | Silverbrook Research Pty Ltd | Injet printhead with thermal bend arm exposed to ink flow |
US5965410A (en) * | 1997-09-02 | 1999-10-12 | Caliper Technologies Corp. | Electrical current for controlling fluid parameters in microchannels |
EP1042061A1 (en) * | 1997-12-24 | 2000-10-11 | Cepheid | Integrated fluid manipulation cartridge |
US6287776B1 (en) * | 1998-02-02 | 2001-09-11 | Signature Bioscience, Inc. | Method for detecting and classifying nucleic acid hybridization |
US6184040B1 (en) * | 1998-02-12 | 2001-02-06 | Polaroid Corporation | Diagnostic assay system and method |
US6200531B1 (en) * | 1998-05-11 | 2001-03-13 | Igen International, Inc. | Apparatus for carrying out electrochemiluminescence test measurements |
US20050244954A1 (en) * | 1998-06-23 | 2005-11-03 | Blackburn Gary F | Binding acceleration techniques for the detection of analytes |
US6761816B1 (en) * | 1998-06-23 | 2004-07-13 | Clinical Micro Systems, Inc. | Printed circuit boards with monolayers and capture ligands |
US6494614B1 (en) * | 1998-07-27 | 2002-12-17 | Battelle Memorial Institute | Laminated microchannel devices, mixing units and method of making same |
US5936730A (en) * | 1998-09-08 | 1999-08-10 | Motorola, Inc. | Bio-molecule analyzer with detector array and filter device |
US6116717A (en) * | 1998-09-15 | 2000-09-12 | Lexmark International, Inc. | Method and apparatus for customized control of a print cartridge |
US6203683B1 (en) * | 1998-11-09 | 2001-03-20 | Princeton University | Electrodynamically focused thermal cycling device |
US6638760B1 (en) * | 1998-11-25 | 2003-10-28 | Pe Corporation (Ny) | Method and apparatus for flow-through hybridization |
US20020177135A1 (en) * | 1999-07-27 | 2002-11-28 | Doung Hau H. | Devices and methods for biochip multiplexing |
US20040053290A1 (en) * | 2000-01-11 | 2004-03-18 | Terbrueggen Robert Henry | Devices and methods for biochip multiplexing |
US6878540B2 (en) * | 1999-06-25 | 2005-04-12 | Cepheid | Device for lysing cells, spores, or microorganisms |
US6453431B1 (en) * | 1999-07-01 | 2002-09-17 | International Business Machines Corporation | System technique for detecting soft errors in statically coupled CMOS logic |
US7078167B2 (en) * | 1999-09-17 | 2006-07-18 | Agilent Technologies, Inc. | Arrays having background features and methods for using the same |
US6699384B1 (en) * | 1999-09-21 | 2004-03-02 | Battelle Memorial Institute | Compact electrochemical sensor system and method for field testing for metals in saliva or other fluids |
WO2001026813A2 (en) * | 1999-10-08 | 2001-04-19 | Micronics, Inc. | Microfluidics without electrically of mechanically operated pumps |
US6576460B1 (en) * | 1999-10-28 | 2003-06-10 | Cornell Research Foundation, Inc. | Filtration-detection device and method of use |
US6553844B2 (en) * | 1999-10-29 | 2003-04-29 | Metasensors, Inc. | Property-independent volumetric flowmeter and sonic velocimeter |
US6867851B2 (en) * | 1999-11-04 | 2005-03-15 | Regents Of The University Of Minnesota | Scanning of biological samples |
US6875619B2 (en) * | 1999-11-12 | 2005-04-05 | Motorola, Inc. | Microfluidic devices comprising biochannels |
CA2396320A1 (en) * | 2000-01-11 | 2001-07-19 | Maxygen, Inc. | Integrated systems and methods for diversity generation and screening |
WO2001055704A1 (en) * | 2000-01-31 | 2001-08-02 | Board Of Regents, The University Of Texas System | System for transferring fluid samples through a sensor array |
JP3871846B2 (en) * | 2000-03-10 | 2007-01-24 | 日立ソフトウエアエンジニアリング株式会社 | Hybridization reaction detection method and detection apparatus |
US7867763B2 (en) * | 2004-01-25 | 2011-01-11 | Fluidigm Corporation | Integrated chip carriers with thermocycler interfaces and methods of using the same |
CH695166A5 (en) * | 2000-04-25 | 2005-12-30 | Sensirion Ag | Method and apparatus for measuring the flow of a liquid. |
EP1297179B1 (en) * | 2000-05-12 | 2008-09-24 | Caliper Life Sciences, Inc. | Detection of nucleic acid hybridization by fluorescence polarization |
WO2001089696A2 (en) * | 2000-05-24 | 2001-11-29 | Micronics, Inc. | Microfluidic concentration gradient loop |
US8071051B2 (en) * | 2004-05-14 | 2011-12-06 | Honeywell International Inc. | Portable sample analyzer cartridge |
US7351376B1 (en) * | 2000-06-05 | 2008-04-01 | California Institute Of Technology | Integrated active flux microfluidic devices and methods |
EP1356088A2 (en) * | 2000-06-07 | 2003-10-29 | Baylor College of Medicine | Compositions and methods for array-based nucleic acid hybridization |
CA2311622A1 (en) * | 2000-06-15 | 2001-12-15 | Moussa Hoummady | Sub-nanoliter liquid drop dispensing system and method therefor |
KR100481305B1 (en) * | 2000-07-21 | 2005-04-07 | 박용덕 | Apparatus for controling a door using a mobile communications system |
FR2812306B1 (en) * | 2000-07-28 | 2005-01-14 | Gabriel Festoc | POLYMERSIS CHAIN AMPLIFICATION SYSTEM OF TARGET NUCLEIC SEQUENCES |
NL1016298C2 (en) * | 2000-09-29 | 2002-04-03 | Sgt Exploitatie Bv | Vial, method for using a vial for analysis on a sample, as well as a system for performing the method according to the invention using a vial according to the invention. |
WO2002028531A1 (en) * | 2000-10-06 | 2002-04-11 | Protasis Corporation | Fluid separation conduit cartridge with encryption capability |
US6827095B2 (en) * | 2000-10-12 | 2004-12-07 | Nanostream, Inc. | Modular microfluidic systems |
JP5416326B2 (en) * | 2000-10-31 | 2014-02-12 | ヒタチ ケミカル リサーチ センター インコーポレイテッド | Collection and use of nuclear mRNA |
US7378280B2 (en) * | 2000-11-16 | 2008-05-27 | California Institute Of Technology | Apparatus and methods for conducting assays and high throughput screening |
US20020094528A1 (en) * | 2000-11-29 | 2002-07-18 | Salafsky Joshua S. | Method and apparatus using a surface-selective nonlinear optical technique for detection of probe-target interations |
US6382254B1 (en) * | 2000-12-12 | 2002-05-07 | Eastman Kodak Company | Microfluidic valve and method for controlling the flow of a liquid |
US7157232B2 (en) * | 2000-12-13 | 2007-01-02 | The Regents Of The University Of California | Method to detect the end-point for PCR DNA amplification using an ionically labeled probe and measuring impedance change |
US20020160363A1 (en) * | 2001-01-31 | 2002-10-31 | Mcdevitt John T. | Magnetic-based placement and retention of sensor elements in a sensor array |
CA2437558A1 (en) * | 2001-01-31 | 2002-08-08 | The Board Of Regents Of The University Of Texas System | Method and apparatus for the confinement of materials in a micromachined chemical sensor array |
US6386219B1 (en) * | 2001-02-01 | 2002-05-14 | Agilent Technologies, Inc. | Fluid handling system and method of manufacture |
EP1360479A4 (en) * | 2001-02-15 | 2005-03-23 | Caliper Life Sciences Inc | Methods and systems for enhanced delivery of electrical currents to fluidic systems |
US20020165675A1 (en) * | 2001-03-03 | 2002-11-07 | Golovlev Valeri V. | Method and microelectronic device for multi-site molecule detection |
US7225807B2 (en) * | 2001-03-15 | 2007-06-05 | Creare Incorporated | Systems and methods for aerosol delivery of agents |
DE10114540A1 (en) * | 2001-03-21 | 2002-10-02 | Francotyp Postalia Ag | Consumption module for an electronic device |
US7323140B2 (en) * | 2001-03-28 | 2008-01-29 | Handylab, Inc. | Moving microdroplets in a microfluidic device |
US20020142318A1 (en) * | 2001-03-30 | 2002-10-03 | Cattell Herbert F. | Chemical array reading |
GB0110501D0 (en) * | 2001-04-30 | 2001-06-20 | Secr Defence Brit | Amplification process |
WO2002089972A1 (en) * | 2001-05-03 | 2002-11-14 | Commissariat A L'energie Atomique | Microfluidic device for analyzing nucleic acids and/or proteins, methods of preparation and uses thereof |
US6573734B2 (en) * | 2001-05-08 | 2003-06-03 | The Board Of Trustees Of The University Of Illinois | Integrated thin film liquid conductivity sensor |
US20050009101A1 (en) * | 2001-05-17 | 2005-01-13 | Motorola, Inc. | Microfluidic devices comprising biochannels |
US7214300B2 (en) * | 2001-06-04 | 2007-05-08 | Epocal Inc. | Integrated electrokinetic devices and methods of manufacture |
WO2002099410A1 (en) * | 2001-06-04 | 2002-12-12 | Aclara Biosciences, Inc. | Sensor device and method for indicating oxygen consumption |
US20030015425A1 (en) * | 2001-06-20 | 2003-01-23 | Coventor Inc. | Microfluidic system including a virtual wall fluid interface port for interfacing fluids with the microfluidic system |
US20030186222A1 (en) | 2001-06-27 | 2003-10-02 | Paul John H. | Rapid detection of enteroviruses in environmental samples by NASBA |
US20030032172A1 (en) * | 2001-07-06 | 2003-02-13 | The Regents Of The University Of California | Automated nucleic acid assay system |
FR2827199B1 (en) * | 2001-07-10 | 2004-07-09 | Centre Nat Rech Scient | PROCESS AND MACHINE FOR THE EX SITU MANUFACTURE OF LOW AND MEDIUM INTEGRATION BIOPE NETWORKS |
DE10133844B4 (en) * | 2001-07-18 | 2006-08-17 | Micronas Gmbh | Method and device for detecting analytes |
EP1439910A2 (en) * | 2001-07-26 | 2004-07-28 | Motorola, Inc. | System and methods for mixing within a microfluidic device |
US6726820B1 (en) * | 2001-09-19 | 2004-04-27 | Applera Corporation | Method of separating biomolecule-containing samples with a microdevice with integrated memory |
US6995841B2 (en) * | 2001-08-28 | 2006-02-07 | Rice University | Pulsed-multiline excitation for color-blind fluorescence detection |
US7075162B2 (en) | 2001-08-30 | 2006-07-11 | Fluidigm Corporation | Electrostatic/electrostrictive actuation of elastomer structures using compliant electrodes |
DE10145701A1 (en) * | 2001-09-17 | 2003-04-10 | Infineon Technologies Ag | Fluorescence biosensor chip and fluorescence biosensor chip arrangement |
US6969843B1 (en) * | 2001-10-19 | 2005-11-29 | Beach James M | Light standard for microscopy |
EP1442142A4 (en) | 2001-10-19 | 2006-11-15 | Proligo Llc | Nucleic acid probes and methods to detect and/or quantify nucleic acid analytes |
US20030175947A1 (en) * | 2001-11-05 | 2003-09-18 | Liu Robin Hui | Enhanced mixing in microfluidic devices |
US6622746B2 (en) * | 2001-12-12 | 2003-09-23 | Eastman Kodak Company | Microfluidic system for controlled fluid mixing and delivery |
US20030148391A1 (en) * | 2002-01-24 | 2003-08-07 | Salafsky Joshua S. | Method using a nonlinear optical technique for detection of interactions involving a conformational change |
US8114349B2 (en) * | 2002-01-28 | 2012-02-14 | Qiagen Sciences, Llc | Bio-analysis cartridge tracking and protection mechanism |
AU2003216175A1 (en) * | 2002-02-04 | 2003-09-02 | Colorado School Of Mines | Laminar flow-based separations of colloidal and cellular particles |
US20060164533A1 (en) * | 2002-08-27 | 2006-07-27 | E-Phocus, Inc | Electronic image sensor |
US20040109793A1 (en) * | 2002-02-07 | 2004-06-10 | Mcneely Michael R | Three-dimensional microfluidics incorporating passive fluid control structures |
US7195986B1 (en) * | 2002-03-08 | 2007-03-27 | Caliper Life Sciences, Inc. | Microfluidic device with controlled substrate conductivity |
JP3722367B2 (en) * | 2002-03-19 | 2005-11-30 | ソニー株式会社 | Manufacturing method of solid-state imaging device |
US6639313B1 (en) | 2002-03-20 | 2003-10-28 | Analog Devices, Inc. | Hermetic seals for large optical packages and the like |
US7312085B2 (en) * | 2002-04-01 | 2007-12-25 | Fluidigm Corporation | Microfluidic particle-analysis systems |
US7156484B2 (en) * | 2002-04-12 | 2007-01-02 | Silverbrook Research Pty Ltd | Inkjet printhead with CMOS drive circuitry close to ink supply passage |
US6877528B2 (en) * | 2002-04-17 | 2005-04-12 | Cytonome, Inc. | Microfluidic system including a bubble valve for regulating fluid flow through a microchannel |
US7157274B2 (en) * | 2002-06-24 | 2007-01-02 | Cytonome, Inc. | Method and apparatus for sorting particles |
US20050033525A1 (en) * | 2002-05-21 | 2005-02-10 | Corson John F. | Method and system for computing and applying a user-defined, global, multi-channel background correction to a feature-based data set obtained from reading a microarray |
US7229838B2 (en) * | 2002-07-08 | 2007-06-12 | Innovative Micro Technology | MEMS actuator and method of manufacture for MEMS particle sorting device |
US7214348B2 (en) | 2002-07-26 | 2007-05-08 | Applera Corporation | Microfluidic size-exclusion devices, systems, and methods |
US20040018635A1 (en) * | 2002-07-26 | 2004-01-29 | Peck Bill J. | Fabricating arrays with drop velocity control |
US6777662B2 (en) * | 2002-07-30 | 2004-08-17 | Freescale Semiconductor, Inc. | System, circuit and method providing a dynamic range pixel cell with blooming protection |
US7118676B2 (en) * | 2003-09-04 | 2006-10-10 | Arryx, Inc. | Multiple laminar flow-based particle and cellular separation with laser steering |
US20040197845A1 (en) * | 2002-08-30 | 2004-10-07 | Arjang Hassibi | Methods and apparatus for pathogen detection, identification and/or quantification |
EP1545740A2 (en) * | 2002-09-07 | 2005-06-29 | Arizona Board of Regents | Integrated apparatus and methods for treating liquids |
US7595883B1 (en) * | 2002-09-16 | 2009-09-29 | The Board Of Trustees Of The Leland Stanford Junior University | Biological analysis arrangement and approach therefor |
ITTO20020808A1 (en) * | 2002-09-17 | 2004-03-18 | St Microelectronics Srl | INTEGRATED DNA ANALYSIS DEVICE. |
WO2004027379A2 (en) * | 2002-09-20 | 2004-04-01 | Novus Molecular, Inc. | Methods and devices for active bioassay |
US7521261B2 (en) * | 2002-09-26 | 2009-04-21 | Vanderbilt University | Method for screening molecular interactions |
WO2004037907A2 (en) * | 2002-10-21 | 2004-05-06 | Biosearch Technologies, Inc. | Luminescent metal ion complexes |
TWI324684B (en) * | 2002-10-25 | 2010-05-11 | Nat Univ Tsing Hua | Micro-array system for micro amount reaction |
US20040086872A1 (en) * | 2002-10-31 | 2004-05-06 | Childers Winthrop D. | Microfluidic system for analysis of nucleic acids |
US7932098B2 (en) * | 2002-10-31 | 2011-04-26 | Hewlett-Packard Development Company, L.P. | Microfluidic system utilizing thin-film layers to route fluid |
US7264723B2 (en) * | 2002-11-01 | 2007-09-04 | Sandia Corporation | Dialysis on microchips using thin porous polymer membranes |
EP1419818B1 (en) * | 2002-11-14 | 2013-10-30 | Boehringer Ingelheim microParts GmbH | Device for sequential transport of liquids by capillary forces |
US6755509B2 (en) * | 2002-11-23 | 2004-06-29 | Silverbrook Research Pty Ltd | Thermal ink jet printhead with suspended beam heater |
JP3624950B2 (en) * | 2002-11-26 | 2005-03-02 | セイコーエプソン株式会社 | ink cartridge |
US20040115794A1 (en) | 2002-12-12 | 2004-06-17 | Affymetrix, Inc. | Methods for detecting transcriptional factor binding sites |
EP1570043B1 (en) * | 2002-12-12 | 2013-07-24 | Novartis Vaccines and Diagnostics, Inc. | Device and method for in-line blood testing using biochips |
US20050042639A1 (en) * | 2002-12-20 | 2005-02-24 | Caliper Life Sciences, Inc. | Single molecule amplification and detection of DNA length |
CA2511389C (en) * | 2002-12-26 | 2016-10-18 | Meso Scale Technologies, Llc. | Assay cartridges and methods of using the same |
US20040188648A1 (en) * | 2003-01-15 | 2004-09-30 | California Institute Of Technology | Integrated surface-machined micro flow controller method and apparatus |
KR20050118668A (en) * | 2003-01-21 | 2005-12-19 | 마이크로닉스 인코포레이티드. | Method and system for microfluidic manipulation, amplification and analysis of fluids, for example, bacteria assays and antiglobulin testing |
US20060210984A1 (en) * | 2003-03-03 | 2006-09-21 | Jeremy Lambert | Use of nucleic acid mimics for internal reference and calibration in a flow cell microarray binding assay |
SE0300823D0 (en) * | 2003-03-23 | 2003-03-23 | Gyros Ab | Preloaded Microscale Devices |
US6986649B2 (en) * | 2003-04-09 | 2006-01-17 | Motorola, Inc. | Micropump with integrated pressure sensor |
US7435381B2 (en) * | 2003-05-29 | 2008-10-14 | Siemens Healthcare Diagnostics Inc. | Packaging of microfluidic devices |
US7309467B2 (en) * | 2003-06-24 | 2007-12-18 | Hewlett-Packard Development Company, L.P. | Fluidic MEMS device |
US20050064465A1 (en) * | 2003-07-02 | 2005-03-24 | Caliper Life Sciences, Inc. | Continuous and non-continuous flow bioreactor |
US20050019951A1 (en) * | 2003-07-14 | 2005-01-27 | Gjerde Douglas T. | Method and device for extracting an analyte |
GB0321158D0 (en) * | 2003-09-10 | 2003-10-08 | Central Research Lab Ltd | Apparatus and method for handling cells,embryos or oocytes |
US20050112634A1 (en) * | 2003-09-19 | 2005-05-26 | Woudenberg Timothy M. | High density sequence detection methods and apparatus |
US8277760B2 (en) * | 2003-09-19 | 2012-10-02 | Applied Biosystems, Llc | High density plate filler |
US7811443B2 (en) * | 2003-10-16 | 2010-10-12 | The Regents Of The University Of California | Microfluidic dynamic vapor control system |
NL1024578C2 (en) * | 2003-10-21 | 2005-04-22 | Univ Delft Tech | Device for carrying out a reaction. |
WO2005047545A2 (en) * | 2003-11-04 | 2005-05-26 | Applera Corporation | Microarray controls |
US20050095602A1 (en) * | 2003-11-04 | 2005-05-05 | West Jason A. | Microfluidic integrated microarrays for biological detection |
US7444005B2 (en) * | 2003-11-04 | 2008-10-28 | Becton, Dickinson And Company | Apparatus and method for using optical mouse engine to determine speed, direction, position of scanned device and to obtain quantitative or qualitative data from same |
US7695952B2 (en) * | 2003-11-07 | 2010-04-13 | Nanosphere, Inc. | Disposable sample processing module for detecting nucleic acids |
EP1716249A2 (en) * | 2003-12-31 | 2006-11-02 | Applera Corporation, Applied Biosystems Group | Quantitative amplification and detection of small numbers of target polynucleotides |
US7526944B2 (en) * | 2004-01-07 | 2009-05-05 | Ashok Sabata | Remote monitoring of pipelines using wireless sensor network |
US7448734B2 (en) * | 2004-01-21 | 2008-11-11 | Silverbrook Research Pty Ltd | Inkjet printer cartridge with pagewidth printhead |
US20050176135A1 (en) * | 2004-02-06 | 2005-08-11 | Brian Jones | Cassette for isolation, amplification and identification of DNA or protein and method of use |
US20060094046A1 (en) * | 2004-02-11 | 2006-05-04 | Arie Abo | Compositions and methods relating to angiogenesis and tumorigenesis |
US7461560B2 (en) * | 2005-03-28 | 2008-12-09 | Microstrain, Inc. | Strain gauge with moisture barrier and self-testing circuit |
US7796266B2 (en) * | 2004-04-30 | 2010-09-14 | Kimberly-Clark Worldwide, Inc. | Optical detection system using electromagnetic radiation to detect presence or quantity of analyte |
US9101302B2 (en) * | 2004-05-03 | 2015-08-11 | Abbott Diabetes Care Inc. | Analyte test device |
JP4683538B2 (en) * | 2004-05-06 | 2011-05-18 | セイコーインスツル株式会社 | Analysis system and analysis method including microchip for analysis |
US7694694B2 (en) * | 2004-05-10 | 2010-04-13 | The Aerospace Corporation | Phase-change valve apparatuses |
TWI291025B (en) | 2004-06-29 | 2007-12-11 | Univ Nat Cheng Kung | An integral micro-dialysis electrophoresis chip having on-line labeling function and the analysis method thereof |
US7134319B2 (en) * | 2004-08-12 | 2006-11-14 | Honeywell International Inc. | Acoustic wave sensor with reduced condensation and recovery time |
EP1817573A4 (en) * | 2004-10-18 | 2010-02-10 | Univ Macquarie | Fluorescence detection |
WO2006054238A2 (en) * | 2004-11-16 | 2006-05-26 | Koninklijke Philips Electronics N. V. | Microfluidic device |
US7785868B2 (en) * | 2004-12-02 | 2010-08-31 | Microfluidic Systems, Inc. | Apparatus to automatically lyse a sample |
JP4455306B2 (en) * | 2004-12-13 | 2010-04-21 | キヤノン株式会社 | Biochemical treatment method |
US20060153745A1 (en) * | 2005-01-11 | 2006-07-13 | Applera Corporation | Fluid processing device for oligonucleotide synthesis and analysis |
US8057756B2 (en) * | 2005-01-28 | 2011-11-15 | Parker-Hannifin Corporation | Sampling probe, gripper and interface for laboratory sample management systems |
GB2438768A (en) * | 2005-02-15 | 2007-12-05 | Univ Singapore | Microfluidics package and method of fabricating the same |
CA2598513A1 (en) * | 2005-02-25 | 2006-08-31 | Inverness Medical Switzerland Gmbh | Fluidic gating device |
EP2597472A3 (en) * | 2005-04-01 | 2014-03-05 | Konica Minolta Medical & Graphic, Inc. | Micro integrated analysis system, testing chip, and testing method |
US7887156B2 (en) * | 2005-04-25 | 2011-02-15 | Ulvac, Inc. | Integral printhead assembly |
GB0508983D0 (en) * | 2005-05-03 | 2005-06-08 | Oxford Gene Tech Ip Ltd | Cell analyser |
US7738086B2 (en) * | 2005-05-09 | 2010-06-15 | The Trustees Of Columbia University In The City Of New York | Active CMOS biosensor chip for fluorescent-based detection |
WO2006122311A2 (en) * | 2005-05-11 | 2006-11-16 | The Trustees Of The University Of Pennsylvania | Microfluidic chip |
EP1885646A1 (en) * | 2005-05-12 | 2008-02-13 | STMicroelectronics S.r.l. | Microfluidic device with integrated micropump, in particular biochemical microreactor, and manufacturing method thereof |
WO2006133101A2 (en) * | 2005-06-03 | 2006-12-14 | Trans-Dermal Patents Company, Llc | Agent delivery system |
WO2007002579A2 (en) * | 2005-06-23 | 2007-01-04 | Bioveris Corporation | Assay cartridges and methods for point of care instruments |
US8288151B2 (en) * | 2005-06-29 | 2012-10-16 | Canon Kabushiki Kaisha | Biochemical reaction cassette |
WO2007005974A2 (en) * | 2005-07-01 | 2007-01-11 | Honeywell International, Inc. | A flow metered analyzer |
KR100672690B1 (en) * | 2005-08-03 | 2007-01-22 | 동부일렉트로닉스 주식회사 | Method for manufacturing of cmos image sensor |
US7731910B2 (en) * | 2005-08-05 | 2010-06-08 | Hewlett-Packard Development Company, L.P. | Microfluidic mixing assembly |
WO2007033385A2 (en) * | 2005-09-13 | 2007-03-22 | Fluidigm Corporation | Microfluidic assay devices and methods |
ES2338368T3 (en) * | 2005-10-12 | 2010-05-06 | Allergan, Inc. | MOLECULAR OR SUBCELLULAR INTERACTIVITY TESTS USING DESPOLARIZATION AFTER RESONANCE TRANSFER (DARET). |
US20070081920A1 (en) * | 2005-10-12 | 2007-04-12 | Murphy R S | Semi-disposable optoelectronic rapid diagnostic test system |
US20070116607A1 (en) * | 2005-11-23 | 2007-05-24 | Pharmacom Microlelectronics, Inc. | Microsystems that integrate three-dimensional microarray and multi-layer microfluidics for combinatorial detection of bioagent at single molecule level |
US8068991B2 (en) * | 2005-11-30 | 2011-11-29 | The Invention Science Fund I, Llc | Systems and methods for transmitting pathogen related information and responding |
US20080241910A1 (en) * | 2007-03-27 | 2008-10-02 | Searete Llc, A Limited Liability Corporation Of The State Of Delaware | Devices for pathogen detection |
EP1971861A4 (en) * | 2005-12-21 | 2014-10-22 | Samsung Electronics Co Ltd | Bio memory disc and bio memory disk drive apparatus, and assay method using the same |
EP1963817A2 (en) * | 2005-12-22 | 2008-09-03 | Honeywell International Inc. | Portable sample analyzer cartridge |
US8703445B2 (en) * | 2005-12-29 | 2014-04-22 | Abbott Point Of Care Inc. | Molecular diagnostics amplification system and methods |
EP1987344A1 (en) | 2006-02-06 | 2008-11-05 | STMicroelectronics S.r.l. | Nucleic acid analysis chip integrating a waveguide and optical apparatus for the inspection of nucleic acid probes |
US7815868B1 (en) * | 2006-02-28 | 2010-10-19 | Fluidigm Corporation | Microfluidic reaction apparatus for high throughput screening |
JP4770530B2 (en) * | 2006-03-13 | 2011-09-14 | 株式会社デンソー | Capacitive humidity sensor |
EP2007905B1 (en) * | 2006-03-15 | 2012-08-22 | Micronics, Inc. | Integrated nucleic acid assays |
US20100120132A1 (en) * | 2006-03-31 | 2010-05-13 | Intel Corporation | Bioassays by direct optical detection of nanoparticles |
US8232091B2 (en) * | 2006-05-17 | 2012-07-31 | California Institute Of Technology | Thermal cycling system |
US20070280857A1 (en) * | 2006-06-02 | 2007-12-06 | Applera Corporation | Devices and Methods for Positioning Dried Reagent In Microfluidic Devices |
US8637436B2 (en) * | 2006-08-24 | 2014-01-28 | California Institute Of Technology | Integrated semiconductor bioarray |
EP2029772B1 (en) * | 2006-06-08 | 2020-03-18 | Koninklijke Philips N.V. | Microelectronic sensor device for dna detection |
WO2007148358A1 (en) * | 2006-06-23 | 2007-12-27 | Stmicroelectronics S.R.L. | Assembly of a microfluidic device for analysis of biological material |
US7629124B2 (en) * | 2006-06-30 | 2009-12-08 | Canon U.S. Life Sciences, Inc. | Real-time PCR in micro-channels |
US8048626B2 (en) | 2006-07-28 | 2011-11-01 | California Institute Of Technology | Multiplex Q-PCR arrays |
US7633606B2 (en) * | 2006-08-24 | 2009-12-15 | Microfluidic Systems, Inc. | Integrated airborne substance collection and detection system |
US8173071B2 (en) * | 2006-08-29 | 2012-05-08 | International Business Machines Corporation | Micro-fluidic test apparatus and method |
US8187541B2 (en) * | 2006-09-18 | 2012-05-29 | California Institute Of Technology | Apparatus for detecting target molecules and related methods |
WO2008147382A1 (en) * | 2006-09-27 | 2008-12-04 | Micronics, Inc. | Integrated microfluidic assay devices and methods |
EP2091647A2 (en) * | 2006-11-14 | 2009-08-26 | Handylab, Inc. | Microfluidic system for amplifying and detecting polynucleotides in parallel |
US20090186034A1 (en) * | 2006-12-19 | 2009-07-23 | Genetech, Inc. | Gene expression markers for inflammatory bowel disease |
WO2008089449A2 (en) * | 2007-01-19 | 2008-07-24 | Biodot, Inc. | Systems and methods for high speed array printing and hybridization |
EP2125219B1 (en) * | 2007-01-19 | 2016-08-10 | Fluidigm Corporation | High precision microfluidic devices and methods |
US7622783B2 (en) * | 2007-02-14 | 2009-11-24 | Innovative Micro Technology | MEMS thermal actuator and method of manufacture |
WO2008113112A1 (en) * | 2007-03-16 | 2008-09-25 | Cleveland Biosensors Pty Ltd | Stop structure for microfluidic device |
GB2447698A (en) * | 2007-03-23 | 2008-09-24 | Univ Exeter | Fabrication of photonic biosensor arrays |
CN101641150B (en) * | 2007-03-23 | 2014-05-07 | 皇家飞利浦电子股份有限公司 | Integrated microfluidic device with reduced peak power |
US20090227005A1 (en) * | 2007-03-27 | 2009-09-10 | Searete Llc, A Limited Liability Corporation Of The State Of Delaware | Methods for pathogen detection |
EP1977830A1 (en) * | 2007-03-30 | 2008-10-08 | Roche Diagnostics GmbH | Micro-fluidic temperature driven valve |
US8425861B2 (en) * | 2007-04-04 | 2013-04-23 | Netbio, Inc. | Methods for rapid multiplexed amplification of target nucleic acids |
WO2009014792A2 (en) * | 2007-05-11 | 2009-01-29 | The Government Of The United States Of America, As Represented By The Secretary Of The Navy | Electrical detection using confined fluids |
CN101855362A (en) * | 2007-05-18 | 2010-10-06 | 美国政府健康及人类服务部,疾病控制和预防中心 | Primers and probes for the detection of streptococcus pneumoniae |
US20100179069A1 (en) | 2007-07-30 | 2010-07-15 | Gn Biosystems Incorporated | Method and apparatus for conducting high-throughput micro-volume experiments |
WO2009015689A1 (en) * | 2007-07-31 | 2009-02-05 | Telefonaktiebolaget Lm Ericsson (Publ) | All optical batcher banyan switch, batcher switch, banyan switch and contention manager |
WO2009021215A1 (en) * | 2007-08-09 | 2009-02-12 | Celula, Inc. | Methods and devices for correlated, multi-parameter single cell measurements and recovery of remnant biological material |
WO2009035621A1 (en) * | 2007-09-13 | 2009-03-19 | Arryx, Inc. | Methods and apparatuses for sorting objects in forensic dna analysis and medical diagnostics |
US20090107907A1 (en) * | 2007-10-24 | 2009-04-30 | University Of Alaska Fairbanks | Droplet-based digital microdialysis |
CN101873885B (en) | 2007-11-22 | 2013-07-31 | 三星电子株式会社 | Thin film valve device and its controlling apparatus |
US8396701B2 (en) * | 2007-12-04 | 2013-03-12 | Lester F. Ludwig | Software systems for development, control, programming, simulation, and emulation of fixed and reconfigurable lab-on-a-chip devices |
EP2072133A1 (en) * | 2007-12-20 | 2009-06-24 | Koninklijke Philips Electronics N.V. | Multi-compartment device with magnetic particles |
KR101435522B1 (en) * | 2008-01-23 | 2014-09-02 | 삼성전자 주식회사 | Biochip |
DE102008009185A1 (en) * | 2008-02-15 | 2009-09-24 | Siemens Aktiengesellschaft | Apparatus and method for detecting liquids or substances from liquids and use of the apparatus |
EP2263299A2 (en) * | 2008-03-28 | 2010-12-22 | Koninklijke Philips Electronics N.V. | Microfluidic device and method |
ES2352581T3 (en) | 2008-06-02 | 2011-02-21 | Boehringer Ingelheim Microparts Gmbh | STRUCTURE OF MICROFLUIDIC SHEET FOR DOSAGE OF LIQUIDS. |
US7887756B2 (en) | 2008-06-20 | 2011-02-15 | Silverbrook Research Pty Ltd | Microfluidic system comprising mechanically-actuated microfluidic pinch valve |
EP2138587A1 (en) * | 2008-06-23 | 2009-12-30 | Koninklijke Philips Electronics N.V. | Amplification of nucleic acids using temperature zones |
US9724695B2 (en) * | 2008-06-23 | 2017-08-08 | Canon U.S. Life Sciences, Inc. | Systems and methods for amplifying nucleic acids |
US9103785B2 (en) * | 2008-06-25 | 2015-08-11 | Emergence Genomics, Llc | Method and apparatus for melting curve analysis of nucleic acids in microarray format |
US8133451B2 (en) * | 2008-08-28 | 2012-03-13 | Microfluidic Systems, Inc. | Sample preparation apparatus |
AU2009285551B2 (en) * | 2008-08-29 | 2015-10-29 | Lee H. Angros | Multiplexed microscope slide staining apparatus |
US20100056394A1 (en) * | 2008-09-04 | 2010-03-04 | Chung Yuan Christian University | Mini Bio-Reactor |
US8707781B2 (en) * | 2008-09-11 | 2014-04-29 | Nxp, B.V. | Sensor has combined in-plane and parallel-plane configuration |
US20100075340A1 (en) * | 2008-09-22 | 2010-03-25 | Mehdi Javanmard | Electrical Detection Of Biomarkers Using Bioactivated Microfluidic Channels |
US9156010B2 (en) * | 2008-09-23 | 2015-10-13 | Bio-Rad Laboratories, Inc. | Droplet-based assay system |
JP2010076380A (en) | 2008-09-29 | 2010-04-08 | Seiko Epson Corp | Liquid container |
CA2740113C (en) * | 2008-10-10 | 2019-12-24 | The Governing Council Of The University Of Toronto | Hybrid digital and channel microfluidic devices and methods of use thereof |
US20100089135A1 (en) * | 2008-10-10 | 2010-04-15 | Nxp B.V. | Device and method for measuring sensor chips |
US20110203700A1 (en) * | 2008-11-13 | 2011-08-25 | Koninklijke Philips Electronics N.V. | Interfacing an inlet to a capillary channel of a microfluidic system |
US8169006B2 (en) * | 2008-11-29 | 2012-05-01 | Electronics And Telecommunications Research Institute | Bio-sensor chip for detecting target material |
EP2194381B1 (en) * | 2008-12-03 | 2015-12-02 | Roche Diagnostics GmbH | Testing element with combined control and calibration zone |
US7964474B2 (en) * | 2008-12-31 | 2011-06-21 | Stmicroelectronics, Inc. | Use of field oxidation to simplify chamber fabrication in microfluidic devices |
WO2010088288A2 (en) * | 2009-01-28 | 2010-08-05 | Fluidigm Corporation | Determination of copy number differences by amplification |
WO2010101926A2 (en) * | 2009-03-02 | 2010-09-10 | The Johns Hopkins University | Microfluidic system for high-throughput, droplet-based single molecule analysis with low reagent consumption |
AU2010256429B2 (en) * | 2009-06-05 | 2015-09-17 | Integenx Inc. | Universal sample preparation system and use in an integrated analysis system |
US9376713B2 (en) * | 2009-09-23 | 2016-06-28 | The Board Of Trustees Of The University Of Illinois | Label free detection of nucleic acid amplification |
US9353407B2 (en) * | 2009-10-21 | 2016-05-31 | Brandeis University | Methods, kits and reaction mixtures for analyzing single-stranded nucleic acid sequences |
CA2994889C (en) * | 2009-12-07 | 2019-01-22 | Meso Scale Technologies, Llc | Assay cartridges and methods of using the same |
US8500979B2 (en) * | 2009-12-31 | 2013-08-06 | Intel Corporation | Nanogap chemical and biochemical sensors |
AU2011221244B2 (en) * | 2010-02-23 | 2014-02-13 | Rheonix, Inc. | Self-contained biological assay apparatus, methods, and applications |
CA2977845C (en) * | 2010-02-23 | 2020-08-04 | Luminex Corporation | Apparatus and methods for integrated sample preparation, reaction and detection |
US9188585B2 (en) * | 2010-05-13 | 2015-11-17 | Robert Bosch Gmbh | Device and method for indirect modulation of detection environment |
US20110312612A1 (en) * | 2010-06-17 | 2011-12-22 | Geneasys Pty Ltd | Loc device for electrochemiluminescent detection of target sequences with probes between a working electrode and a photosensor |
KR20120063162A (en) * | 2010-12-07 | 2012-06-15 | 삼성전자주식회사 | Gene analysis apparatus and method of analyzing gene using the same |
-
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- 2011-06-01 US US13/150,000 patent/US20110312690A1/en not_active Abandoned
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- 2011-06-01 US US13/149,952 patent/US20110312657A1/en not_active Abandoned
- 2011-06-01 US US13/150,094 patent/US20110312742A1/en not_active Abandoned
- 2011-06-01 US US13/150,214 patent/US20110311413A1/en not_active Abandoned
- 2011-06-01 US US13/150,219 patent/US20110311414A1/en not_active Abandoned
- 2011-06-01 US US13/150,114 patent/US20110312758A1/en not_active Abandoned
- 2011-06-01 US US13/150,170 patent/US20110312781A1/en not_active Abandoned
- 2011-06-01 US US13/150,003 patent/US20110312693A1/en not_active Abandoned
- 2011-06-01 US US13/150,238 patent/US20110312810A1/en not_active Abandoned
- 2011-06-01 US US13/150,045 patent/US20110312708A1/en not_active Abandoned
- 2011-06-01 US US13/149,985 patent/US20110312683A1/en not_active Abandoned
- 2011-06-01 US US13/149,993 patent/US20110312686A1/en not_active Abandoned
- 2011-06-01 US US13/149,894 patent/US20110312624A1/en active Pending
- 2011-06-01 US US13/150,130 patent/US20110312768A1/en not_active Abandoned
- 2011-06-01 US US13/150,175 patent/US20110312601A1/en not_active Abandoned
- 2011-06-01 US US13/149,950 patent/US20110312655A1/en not_active Abandoned
- 2011-06-01 US US13/149,907 patent/US20110312632A1/en not_active Abandoned
- 2011-06-01 US US13/150,266 patent/US20110312835A1/en not_active Abandoned
- 2011-06-01 US US13/150,137 patent/US20110312081A1/en not_active Abandoned
- 2011-06-01 US US13/150,073 patent/US20110312729A1/en not_active Abandoned
- 2011-06-01 US US13/149,929 patent/US20110311411A1/en not_active Abandoned
- 2011-06-01 US US13/149,898 patent/US8349277B2/en not_active Expired - Fee Related
- 2011-06-01 US US13/150,134 patent/US20110312770A1/en not_active Abandoned
- 2011-06-01 US US13/150,220 patent/US20110312610A1/en not_active Abandoned
- 2011-06-01 US US13/150,191 patent/US20110312606A1/en not_active Abandoned
- 2011-06-01 US US13/150,119 patent/US20110312763A1/en not_active Abandoned
- 2011-06-01 US US13/150,004 patent/US20110312694A1/en not_active Abandoned
- 2011-06-01 US US13/150,093 patent/US20110312741A1/en not_active Abandoned
- 2011-06-01 US US13/150,068 patent/US20110312577A1/en not_active Abandoned
- 2011-06-01 US US13/149,895 patent/US20110312625A1/en not_active Abandoned
- 2011-06-01 US US13/149,943 patent/US20110312076A1/en not_active Abandoned
- 2011-06-01 US US13/150,123 patent/US20110312765A1/en not_active Abandoned
- 2011-06-01 US US13/150,148 patent/US20110312843A1/en not_active Abandoned
- 2011-06-01 WO PCT/AU2011/000678 patent/WO2011156855A1/en active Application Filing
- 2011-06-01 US US13/150,080 patent/US20110312734A1/en not_active Abandoned
- 2011-06-01 WO PCT/AU2011/000680 patent/WO2011156857A1/en active Application Filing
- 2011-06-01 WO PCT/AU2011/000664 patent/WO2011156841A1/en active Application Filing
- 2011-06-01 US US13/150,056 patent/US20110312718A1/en not_active Abandoned
- 2011-06-01 US US13/150,216 patent/US20110312608A1/en not_active Abandoned
- 2011-06-01 US US13/150,088 patent/US20110312739A1/en not_active Abandoned
- 2011-06-01 US US13/150,131 patent/US8354074B2/en not_active Expired - Fee Related
- 2011-06-01 US US13/150,012 patent/US20110312696A1/en not_active Abandoned
- 2011-06-01 US US13/150,058 patent/US20110312720A1/en not_active Abandoned
- 2011-06-01 US US13/150,059 patent/US20110312575A1/en not_active Abandoned
- 2011-06-01 US US13/150,165 patent/US20110312778A1/en not_active Abandoned
- 2011-06-01 US US13/150,033 patent/US20110312077A1/en not_active Abandoned
- 2011-06-01 US US13/150,201 patent/US8398938B2/en not_active Expired - Fee Related
- 2011-06-01 US US13/150,018 patent/US20110312697A1/en not_active Abandoned
- 2011-06-01 US US13/150,146 patent/US20110311394A1/en not_active Abandoned
- 2011-06-01 US US13/150,051 patent/US20110312714A1/en not_active Abandoned
- 2011-06-01 US US13/150,108 patent/US20110312752A1/en not_active Abandoned
- 2011-06-01 US US13/149,971 patent/US20110312674A1/en not_active Abandoned
- 2011-06-01 US US13/150,237 patent/US20110312809A1/en not_active Abandoned
- 2011-06-01 US US13/150,213 patent/US20110312800A1/en not_active Abandoned
- 2011-06-01 WO PCT/AU2011/000666 patent/WO2011156843A1/en active Application Filing
- 2011-06-01 US US13/150,255 patent/US20110312825A1/en not_active Abandoned
- 2011-06-01 US US13/149,911 patent/US20110312634A1/en not_active Abandoned
- 2011-06-01 US US13/149,918 patent/US20110312638A1/en not_active Abandoned
- 2011-06-01 US US13/149,983 patent/US20110312681A1/en not_active Abandoned
- 2011-06-01 US US13/150,006 patent/US20110312538A1/en not_active Abandoned
- 2011-06-01 US US13/149,978 patent/US20110312678A1/en not_active Abandoned
- 2011-06-01 US US13/150,100 patent/US20110312078A1/en not_active Abandoned
- 2011-06-01 US US13/150,204 patent/US20110312795A1/en not_active Abandoned
- 2011-06-01 US US13/150,089 patent/US20110312740A1/en not_active Abandoned
- 2011-06-01 US US13/150,014 patent/US20110312561A1/en not_active Abandoned
- 2011-06-01 US US13/150,019 patent/US20110312698A1/en not_active Abandoned
- 2011-06-01 US US13/150,063 patent/US20110312724A1/en not_active Abandoned
- 2011-06-01 US US13/149,899 patent/US20110312546A1/en not_active Abandoned
- 2011-06-01 US US13/150,239 patent/US20110312811A1/en not_active Abandoned
- 2011-06-01 US US13/149,897 patent/US20110312626A1/en not_active Abandoned
- 2011-06-01 US US13/150,040 patent/US20110312705A1/en not_active Abandoned
- 2011-06-01 US US13/150,087 patent/US20110312615A1/en not_active Abandoned
- 2011-06-01 US US13/150,272 patent/US20110308945A1/en not_active Abandoned
- 2011-06-01 US US13/150,075 patent/US20110312731A1/en not_active Abandoned
- 2011-06-01 US US13/150,038 patent/US20110312540A1/en not_active Abandoned
- 2011-06-01 WO PCT/AU2011/000667 patent/WO2011156844A1/en active Application Filing
- 2011-06-01 US US13/150,096 patent/US20110312616A1/en not_active Abandoned
- 2011-06-01 US US13/150,253 patent/US20110312824A1/en not_active Abandoned
- 2011-06-01 US US13/149,931 patent/US20110312641A1/en not_active Abandoned
- 2011-06-01 US US13/150,112 patent/US20110312756A1/en not_active Abandoned
- 2011-06-01 US US13/150,036 patent/US20110312571A1/en not_active Abandoned
- 2011-06-01 US US13/150,240 patent/US20110312812A1/en not_active Abandoned
- 2011-06-01 US US13/149,913 patent/US20110312071A1/en not_active Abandoned
- 2011-06-01 WO PCT/AU2011/000672 patent/WO2011156849A1/en active Application Filing
- 2011-06-01 US US13/150,250 patent/US20110312821A1/en not_active Abandoned
- 2011-06-01 US US13/150,002 patent/US20110312692A1/en not_active Abandoned
- 2011-06-01 US US13/150,118 patent/US20110312762A1/en not_active Abandoned
- 2011-06-01 US US13/150,143 patent/US20110312842A1/en not_active Abandoned
- 2011-06-01 US US13/149,937 patent/US20110312647A1/en not_active Abandoned
- 2011-06-01 US US13/150,062 patent/US20110312723A1/en not_active Abandoned
- 2011-06-01 US US13/149,968 patent/US20110312671A1/en not_active Abandoned
- 2011-06-01 US US13/150,241 patent/US20110312813A1/en not_active Abandoned
- 2011-06-01 US US13/149,903 patent/US20110312547A1/en not_active Abandoned
- 2011-06-01 US US13/149,951 patent/US20110312656A1/en not_active Abandoned
- 2011-06-01 US US13/150,057 patent/US8383065B2/en not_active Expired - Fee Related
- 2011-06-01 US US13/150,262 patent/US20110312831A1/en not_active Abandoned
- 2011-06-01 US US13/149,908 patent/US20110312548A1/en not_active Abandoned
- 2011-06-01 US US13/150,097 patent/US20110312744A1/en not_active Abandoned
- 2011-06-01 US US13/149,960 patent/US20110312553A1/en not_active Abandoned
- 2011-06-01 US US13/149,981 patent/US20110312680A1/en not_active Abandoned
- 2011-06-01 US US13/149,975 patent/US20110312677A1/en not_active Abandoned
- 2011-06-01 US US13/150,193 patent/US20110312789A1/en not_active Abandoned
- 2011-06-01 US US13/150,039 patent/US20110312572A1/en not_active Abandoned
- 2011-06-01 TW TW100119255A patent/TW201213798A/en unknown
- 2011-06-01 US US13/150,181 patent/US20110312786A1/en not_active Abandoned
- 2011-06-01 WO PCT/AU2011/000663 patent/WO2011156840A1/en active Application Filing
- 2011-06-01 US US13/150,150 patent/US20110312773A1/en not_active Abandoned
- 2011-06-01 US US13/150,223 patent/US20110312611A1/en not_active Abandoned
- 2011-06-01 US US13/150,011 patent/US20110312695A1/en not_active Abandoned
- 2011-06-01 US US13/149,902 patent/US20110312629A1/en not_active Abandoned
- 2011-06-01 US US13/150,032 patent/US20110312702A1/en not_active Abandoned
- 2011-06-01 US US13/150,079 patent/US20110312733A1/en not_active Abandoned
- 2011-06-01 US US13/150,135 patent/US8383064B2/en not_active Expired - Fee Related
- 2011-06-01 US US13/149,995 patent/US20110312687A1/en not_active Abandoned
- 2011-06-01 US US13/150,229 patent/US20110312803A1/en not_active Abandoned
- 2011-06-01 US US13/149,904 patent/US20110312630A1/en not_active Abandoned
- 2011-06-01 US US13/149,942 patent/US20110312650A1/en not_active Abandoned
- 2011-06-01 US US13/150,132 patent/US8394339B2/en not_active Expired - Fee Related
- 2011-06-01 WO PCT/AU2011/000658 patent/WO2011156835A1/en active Application Filing
- 2011-06-01 US US13/150,234 patent/US20110312856A1/en not_active Abandoned
- 2011-06-01 US US13/150,235 patent/US20110312807A1/en not_active Abandoned
- 2011-06-01 US US13/150,179 patent/US20110312603A1/en not_active Abandoned
- 2011-06-01 US US13/150,125 patent/US20110312069A1/en not_active Abandoned
- 2011-06-01 US US13/150,086 patent/US20110312738A1/en not_active Abandoned
- 2011-06-01 US US13/149,992 patent/US20110312557A1/en not_active Abandoned
- 2011-06-01 US US13/150,072 patent/US20110312580A1/en not_active Abandoned
- 2011-06-01 US US13/150,184 patent/US8425845B2/en not_active Expired - Fee Related
- 2011-06-01 US US13/150,159 patent/US20110312775A1/en not_active Abandoned
- 2011-06-01 US US13/150,140 patent/US20110311393A1/en not_active Abandoned
- 2011-06-01 US US13/149,924 patent/US20110312551A1/en not_active Abandoned
- 2011-06-01 US US13/149,946 patent/US20110312652A1/en not_active Abandoned
- 2011-06-01 US US13/150,104 patent/US20110312749A1/en not_active Abandoned
- 2011-06-01 US US13/150,069 patent/US20110312578A1/en not_active Abandoned
- 2011-06-01 US US13/150,225 patent/US20110312801A1/en not_active Abandoned
- 2011-06-01 US US13/149,932 patent/US20110312642A1/en not_active Abandoned
- 2011-06-01 US US13/149,906 patent/US20110312631A1/en not_active Abandoned
- 2011-06-01 US US13/150,084 patent/US20110312737A1/en not_active Abandoned
- 2011-06-01 US US13/150,113 patent/US20110312757A1/en not_active Abandoned
- 2011-06-01 US US13/150,095 patent/US20110312743A1/en not_active Abandoned
- 2011-06-01 US US13/150,246 patent/US20110312817A1/en not_active Abandoned
- 2011-06-01 US US13/150,061 patent/US8388910B2/en not_active Expired - Fee Related
- 2011-06-01 WO PCT/AU2011/000660 patent/WO2011156837A1/en active Application Filing
- 2011-06-01 US US13/149,933 patent/US20110312643A1/en not_active Abandoned
- 2011-06-01 US US13/150,153 patent/US20120028842A1/en not_active Abandoned
- 2011-06-01 US US13/150,177 patent/US20110312602A1/en not_active Abandoned
- 2011-06-01 US US13/150,141 patent/US20110312082A1/en not_active Abandoned
- 2011-06-01 US US13/150,158 patent/US20110311395A1/en not_active Abandoned
- 2011-06-01 US US13/150,066 patent/US20110312576A1/en not_active Abandoned
- 2011-06-01 US US13/149,962 patent/US20110312666A1/en not_active Abandoned
- 2011-06-01 US US13/149,891 patent/US20110312841A1/en not_active Abandoned
- 2011-06-01 US US13/150,172 patent/US20110312782A1/en not_active Abandoned
- 2011-06-01 US US13/150,180 patent/US20110312785A1/en not_active Abandoned
- 2011-06-01 US US13/149,936 patent/US20110312646A1/en not_active Abandoned
- 2011-06-01 US US13/149,966 patent/US20110312670A1/en not_active Abandoned
- 2011-06-01 US US13/150,127 patent/US20120053088A1/en not_active Abandoned
- 2011-06-01 US US13/150,120 patent/US20110311418A1/en not_active Abandoned
- 2011-06-01 US US13/149,965 patent/US20110312669A1/en not_active Abandoned
- 2011-06-01 US US13/150,257 patent/US20110312827A1/en not_active Abandoned
- 2011-06-01 US US13/149,996 patent/US20110312688A1/en not_active Abandoned
- 2011-06-01 US US13/150,009 patent/US20110312560A1/en not_active Abandoned
- 2011-06-01 US US13/149,953 patent/US20110312658A1/en not_active Abandoned
- 2011-06-01 US US13/149,989 patent/US20110312684A1/en not_active Abandoned
- 2011-06-01 US US13/150,192 patent/US20110312607A1/en not_active Abandoned
- 2011-06-01 US US13/150,007 patent/US20110312559A1/en not_active Abandoned
- 2011-06-01 US US13/150,221 patent/US20120004145A1/en not_active Abandoned
- 2011-06-01 WO PCT/AU2011/000679 patent/WO2011156856A1/en active Application Filing
- 2011-06-01 US US13/150,248 patent/US20110312819A1/en not_active Abandoned
- 2011-06-01 US US13/149,991 patent/US20110312556A1/en not_active Abandoned
- 2011-06-01 US US13/150,106 patent/US20110312751A1/en not_active Abandoned
- 2011-06-01 US US13/150,166 patent/US20110312079A1/en not_active Abandoned
- 2011-06-01 US US13/150,209 patent/US20110309275A1/en not_active Abandoned
- 2011-06-01 US US13/150,210 patent/US20110312798A1/en not_active Abandoned
- 2011-06-01 US US13/149,928 patent/US20110312640A1/en not_active Abandoned
- 2011-06-01 US US13/150,169 patent/US20110312780A1/en not_active Abandoned
- 2011-06-01 US US13/150,162 patent/US20110312598A1/en not_active Abandoned
- 2011-06-01 US US13/150,174 patent/US20110312783A1/en not_active Abandoned
- 2011-06-01 US US13/149,997 patent/US20110312558A1/en not_active Abandoned
- 2011-06-01 US US13/150,020 patent/US20110312564A1/en not_active Abandoned
- 2011-06-01 US US13/149,935 patent/US20110312645A1/en not_active Abandoned
- 2011-06-01 US US13/150,115 patent/US20110312759A1/en not_active Abandoned
- 2011-06-01 US US13/150,233 patent/US20110312806A1/en not_active Abandoned
- 2011-06-01 WO PCT/AU2011/000676 patent/WO2011156853A1/en active Application Filing
- 2011-06-01 US US13/150,064 patent/US8398940B2/en not_active Expired - Fee Related
- 2011-06-01 US US13/150,024 patent/US20110312567A1/en not_active Abandoned
- 2011-06-01 US US13/150,083 patent/US20110312736A1/en not_active Abandoned
- 2011-06-01 US US13/150,185 patent/US20110312787A1/en not_active Abandoned
- 2011-06-01 US US13/150,090 patent/US20110312583A1/en not_active Abandoned
- 2011-06-01 US US13/150,081 patent/US20110312527A1/en not_active Abandoned
-
2012
- 2012-11-26 US US13/685,105 patent/US20130079254A1/en not_active Abandoned
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1995025116A1 (en) * | 1994-03-16 | 1995-09-21 | California Institute Of Technology | Method and apparatus for performing multiple sequential reactions on a matrix |
WO1998041531A2 (en) * | 1997-03-20 | 1998-09-24 | University Of Washington | Solvent for biopolymer synthesis, solvent microdroplets and methods of use |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102649931A (en) * | 2012-05-28 | 2012-08-29 | 上海理工大学 | Preparation method for microarray biochip |
WO2016062788A1 (en) | 2014-10-24 | 2016-04-28 | Ait Austrian Institute Of Technology Gmbh | Microfluidic chip for biological analysis |
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