WO1998053311A2 - Devices and methods for using centripetal acceleration to drive fluid movement in a microfluidics system - Google Patents

Devices and methods for using centripetal acceleration to drive fluid movement in a microfluidics system Download PDF

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Publication number
WO1998053311A2
WO1998053311A2 PCT/US1998/010520 US9810520W WO9853311A2 WO 1998053311 A2 WO1998053311 A2 WO 1998053311A2 US 9810520 W US9810520 W US 9810520W WO 9853311 A2 WO9853311 A2 WO 9853311A2
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WO
WIPO (PCT)
Prior art keywords
capillary
chamber
fluid
platform
rotational speed
Prior art date
Application number
PCT/US1998/010520
Other languages
French (fr)
Other versions
WO1998053311A9 (en
WO1998053311A3 (en
Inventor
Gregory Kellogg
Stephen G. Kieffer-Higgins
Bruce L. Carvalho
Gene A. Davis
John P. Willis
Ted Minior
Laura L. Chapman
Mikayla Kob
Sarah D. Oeltjen
Shari Ommert
Alec Mian
Original Assignee
Gamera Bioscience Corporation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Gamera Bioscience Corporation filed Critical Gamera Bioscience Corporation
Priority to EP98923688A priority Critical patent/EP0983504A2/en
Priority to AU75919/98A priority patent/AU7591998A/en
Priority to JP55068998A priority patent/JP3469585B2/en
Publication of WO1998053311A2 publication Critical patent/WO1998053311A2/en
Publication of WO1998053311A3 publication Critical patent/WO1998053311A3/en
Publication of WO1998053311A9 publication Critical patent/WO1998053311A9/en

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    • Y10T137/1797Heat destructible or fusible
    • Y10T137/1812In fluid flow path
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10TTECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T137/00Fluid handling
    • Y10T137/4456With liquid valves or liquid trap seals
    • Y10T137/4643Liquid valves

Definitions

  • This invention relates to methods and apparatus for performing microanalytic and microsynthetic analyses and procedures.
  • the invention relates to microminiaturization of genetic, biochemical and chemical processes related to analysis, synthesis and purification.
  • the invention provides a microsystem platform and a micromanipulation device to manipulate the platform by rotation, thereby utilizing the centripetal forces resulting from rotation of the platform to motivate fluid movement through microchannels embedded in the microplatform.
  • the microsystem platforms of the invention are also provided having microfluidics components, resistive heating elements, temperature sensing elements, mixing structures, capillary and sacrificial valves, and methods for using these microsystems platforms for performing biological, enzymatic, immunological and chemical assays.
  • a slip ring designed rotor capable of transferring electrical signals to and from the microsystem platforms of the invention is also provided.
  • U.S. Patent No. 4,515,889, issued May 7, 1985 to Klose et al. teach automated mixing and incubating reagents to perform analytical determinations.
  • U.S. Patent No. 4,676,952, issued June 30, 1987 to Edelmann et al. teach a photometric analysis apparatus.
  • U.S. Patent No.4,745,072, issued May 17, 1998 to Ekins discloses immunoassay in biological fluids.
  • U.S. Patent No. 5,061,381, issued October 29, 1991 to Burd discloses a centrifugal rotor for performing blood analyses.
  • U.S. Patent No. 5,122,284, issued June 16, 1992 to Braynin et al. discloses a centrifugal rotor comprising a plurality of peripheral cuvettes.
  • U.S. Patent No. 5,173,193, issued December 22, 1992 to Schembri discloses a centrifugal rotor for delivering a metered amount of a fluid to a receiving chamber on the rotor.
  • U.S. Patent No. 5,409,665 issued April 25, 1995 to Burd discloses a cuvette filling in a centrifuge rotor.
  • U.S. Patent No. 5,472,603, issued December 5, 1995 to Schembri discloses an analytical rotor comprising a capillary passage having an exit duct wherein capillary forces prevent fluid flow at a given rotational speed and permit flow at a higher rotational speed.
  • Renoe et al., 1974 Clin. Chem. 20: 955-960 teach a "minidisc” module for a centrifugal analyzer.
  • Burtis et al, 1975, Clin. Chem. 20: 932-941 teach a method for a dynamic introduction of liquids into a centrifugal analyzer.
  • Fritsche et al, 1975, Clin. Biochem. 8: 240-246 teach enzymatic analysis of blood sugar levels using a centrifugal analyzer.
  • U.S. Patent No. 5,006,749, issued April 9, 1991 to White disclose methods apparatus for using ultrasonic energy to move microminiature elements.
  • microanalytical methods and apparati has been the difficulty in designing systems for moving fluids on microchips through channels and reservoirs having diameters in the 10-100 ⁇ m range.
  • Micro fluidic systems require precise and accurate control of fluid flow and valving to control chemical reactions and analyte detection.
  • Conventional pumping and valving mechanisms have been difficult to incorporate into microscale structures due to inherent conflicts-of-scale. These conflicts of scale arise in part due to the fact that molecular interactions arising out of mechanical components of such components, which are negligible in large
  • This invention provides microsystems platforms as disclosed in co-owned and co-pending U.S. Serial No. 08/761,063, filed December 5, 1996 and incorporated by reference herein. Specifically, this invention provides microfluidics components, resistive heating elements, temperature sensing elements, mixing structures, capillary and sacrificial valves, and methods for using these microsystems platforms for performing biological, enzymatic, immunological and chemical assays.
  • centrifugal rotors and microsystems platforms of the invention that an imprecise amount of a fluid comprising a biological sample can be applied to the rotor or platform and a precise volumetric amount of the biological sample is delivered to a fluid reservoir comprising a reaction vessel or other component of the rotor of platform for performing chemical, biochemical, immunological or other analyses. It is an advantage of the centrifugal rotors and microsystems platforms of the invention that metering of said precise amount of a biological fluid sample, for example, a drop of blood, is provided as an intrinsic property of the metering capillary channel of the rotor or platform, thereby avoiding variability introduced by centripetal metering of the sample into a reaction reservoir.
  • centrifugal rotors and microsystems platforms of the invention that an operator can avoid having to precisely measure an amount of a fluid comprising a biological sample for application to the rotor or microsystem platform, thereby permitting end-users, including consumers, having a lower level of sophistication to use a medically diagnostic or other embodiment of the rotor or microsystem platform of the invention.
  • centrifugal rotors and microsystems platforms of the invention fluid movement into and out of fluid reservoirs on the rotor or platform is precisely determined by displacement of a first fluid, such as biological sample, from a fluid reservoir by a second fluid contained in a second reservoir on the rotor or platform. It is also an advantage of the centrifugal rotors and microsystems platforms of the invention that approximately complete replacement of the volumetric capacity of a first reservoir can be achieved by using fluid displacement as disclosed herein, thereby providing for maximum recovery of a first fluid sample upon displacement by a second fluid, or maximum delivery and replacement of the first fluid by the second fluid. This aspect of the invention is advantageous for providing sequential chemical or biochemical reaction steps wherein mixing of the reagents is not desired.
  • Figures 1 through 3 illustrate the microfluidics array of the microsystem platform described in Example 1.
  • Figures 4 through 6 illustrate the microfluidics array of the microsystem platform described in Example 2.
  • FIGS 7 through 9 illustrate the microfluidics array of the microsystem platform described in Example 3.
  • FIGS 10 through 12 illustrate the microfluidics array of the microsystem platform described in Example 4.
  • Figures 16 through 18 illustrate the microfluidics array of the microsystem platform described in Example 6.
  • Figures 19 through 21 illustrate the microfluidics array of the microsystem platform described in Example 7.
  • Figures 22 through 24 illustrate the microfluidics array of the microsystem platform described in Example 8.
  • FIGS 25 through 27 illustrate the microfluidics array of the microsystem platform described in Example 9.
  • Figure 28 illustrates a microfluidics array of a multiplicity of mixing chambers of the microsystem platform described in Example 8.
  • Figure 29 illustrates the electronic spindle of the invention.
  • Figure 30 illustrates screen-printing of a resistive heater element as described in
  • FIG. 31 illustrates screen-printing of a resistive heater element as described in Example 10.
  • Figure 32 is a graph showing the time dependence of temperature produced at a variety of voltages using a resistive heater element as described in Example 10.
  • Figure 33 is a graph showing the voltage dependence of temperature produced at a variety of voltages using a resistive heater element as described in Example 10.
  • Figure 34 is a graph showing the distance dependence on heating using a resistive heater element as described in Example 10.
  • Figure 35 illustrates screen-printing of a resistive heater element associated with wax valves as described in Example 11.
  • Figure 36 illustrates screen-printing of a resistive heater element associated with wax valves as described in Example 11.
  • Figure 37 illustrates screen-printing of a resistive heater element associated with wax valves as described in Example 11.
  • Figure 38 illustrates the use of screen-printed resistive heater elements to melt wax valves and control fluid flow in a microfluidics array as described in Example 11.
  • Figure 39 is a graph showing that temperature can be cycled using a screen- printed resistive heater element as described in Example 14.
  • the present invention provides centrifugal rotors and microsystems platforms for providing centripetally-motivated fluid micromanipulation.
  • sample will be understood to encompass any fluid, solution or mixture, either isolated or detected as a constituent of a more complex mixture, or synthesized from precursor species.
  • the term “in fluid communication” or “fluidly connected” is intended to define components that are operably interconnected to allow fluid flow between components.
  • the platform comprises a rotatable platform, more preferably a disk, whereby fluid movement on the disk is motivated by centripetal force upon rotation of the disk.
  • a centripetally motivated fluid micromanipulation apparatus is intended to include analytical centrifuges and rotors, microscale centrifugal separation apparati, and most particularly the microsystems platforms and disk handling apparati of International Application No. WO97/21090.
  • the term “microsystems platform” is intended to include centripetally-motivated microfluidics arrays as disclosed in International Application No. WO97/21090.
  • capillary For the purposes of this invention, the terms "capillary”, “microcapillary” and “microchannel” will be understood to be interchangeable and to be constructed of either wetting or non-wetting materials where appropriate.
  • fluid chamber will be understood to mean a defined volume on a rotor or microsystems platform of the invention comprising a fluid.
  • the term "entry port” will be understood to mean a defined volume on a rotor or microsystems platform of the invention comprising a means for applying a fluid to the rotor or platform.
  • capillary junction will be understood to mean a junction of two components wherein one or both of the lateral dimensions of the junction are larger than the corresponding dimensions the capillary.
  • wetting or wettable systems the such junctions are where capillary valving occurs, because fluid flow through the capillaries is stopped at such junctions.
  • non-wetting or non-wettable junctions the exit from the chamber or reservoir is where the capillary junction occurs.
  • capillary junctions are formed when the dimensions of the components change from a small diameter (such as a capillary) to a larger diameter (such as a chamber) in wetting systems, in contrast to non- wettable systems, where capillary junctions form when the dimensions of the components change from a larger diameter (such as a chamber) to a small diameter (such as a capillary).
  • biological sample or “biological fluid sample” will be understood to mean any biologically-derived analytical sample, including but not limited to blood, plasma, serum, lymph, saliva, tears, cerebrospinal fluid, urine, sweat, plant and vegetable extracts, semen, and ascites fluid.
  • air displacement channels will be understood to include ports in the surface of the platform that are contiguous with the components (such as chambers and reservoirs) on the platform, and that comprise vents and microchannels that permit displacement of air from components of the platforms and rotors by fluid movement.
  • capillary action will be understood to mean fluid flow in the absence of rotational motion or centripetal force applied to a fluid on a rotor or platform of the invention.
  • capillary micro valve will be understood to mean a capillary comprising a capillary junction whereby fluid flow is impeded and can be motivated by the application of pressure on a fluid, typically by centripetal force created by rotation of the rotor or platform of the invention.
  • microplatforms of the invention preferably and hereinafter collectively referred to as “disks”; for the purposes of this invention, the terms "microplatform”,
  • microsystems platform and “disk” are considered to be interchangeable), are provided to comprise one or a multiplicity of microsynthetic or microanalytic systems.
  • Such microsynthetic or microanalytic systems in turn comprise combinations of related components as described in further detail herein that are operably interconnected to allow fluid flow between components upon rotation of the disk.
  • These components can be fabricated as described below either integral to the disk or as modules attached to, placed upon, in contact with or embedded in the disk.
  • the invention also comprises a micromanipulation device for manipulating the disks of the invention, wherein the disk is rotated within the device to provide centripetal force to effect fluid flow on the disk.
  • the device provides means for rotating the disk at a controlled rotational velocity, for stopping and starting disk rotation, and advantageously for changing the direction of rotation of the disk.
  • electromechanical means and control means are provided as components of the devices of the invention.
  • User interface means (such as a keypad and a display) are also provided, as further described in International Application WO97/21090.
  • Fluid (including reagents, samples and other liquid components) movement is controlled by centripetal acceleration due to rotation of the platform.
  • the magnitude of centripetal acceleration required for fluid to flow at a rate and under a pressure appropriate for a particular microsystem is determined by factors including but not limited to the effective radius of the platform, the position angle of the structures on the platform with respect to the direction of rotation and the speed of rotation of the platform.
  • the capillary junctions and microvalves of the invention are based on the use of rotationally-induced fluid pressure to overcome capillary forces. Fluids which completely or partially wet the material of the microchannels (or reservoirs, reaction chambers, detection chambers, etc.) which contain them experience a resistance to flow when moving from a microchannel of narrow cross-section to one of larger cross- section, while those fluids which do not wet these materials resist flowing from microchannels (or reservoirs, reaction chambers, detection chambers, etc.) of large cross- section to those with smaller cross-section.
  • This capillary pressure varies inversely with the sizes of the two microchannels (or reservoirs, reaction chambers, detection chambers, etc., or combinations thereof), the surface tension of the fluid, and the contact angle of the fluid on the material of the microchannels (or reservoirs, reaction chambers, detection chambers, etc.) .
  • the details of the cross-sectional shape are not important, but the dependence on cross-sectional dimension results in microchannels of dimension less than 500 ⁇ m exhibit significant capillary pressure.
  • "valve" are fashioned that require the application of a particular pressure on the fluid to induce fluid flow.
  • This pressure is applied in the disks of the invention by rotation of the disk (which has been shown above to vary with the square of the rotational frequency, with the radial position and with the extent of the fluid in the radial direction).
  • capillary valve cross-sectional dimensions as well as the position and extent along the radial direction of the fluid handling components of the microsystem platforms of the invention, capillary valves are formed to release fluid flow in a rotation-dependent manner, using rotation rates of from lOOrpm to several thousand rpm. This arrangement allows complex, multistep fluid processes to be carried out using a pre-determined, mono tonic increase in rotational rate.
  • the theoretical principles underlying the use of capillary junctions and microvalves are disclosed in International Patent Application, Publication No. WO97/ .
  • the instant invention provides microsystems platforms comprising microfluidics components, heating elements, temperature sensing elements, capillary valves, sacrificial valves and a rotor design for transmitting electrical signals to and from the microsystems platforms of the invention.
  • the invention provides fluidics components for capillary metering of precise amounts of a volume of a fluid sample from the application of a less precise volume of a fluid sample at an entry port on the microsystem platform.
  • These embodiments of the invention provide for delivery of precise amounts of a sample such as a biological fluid sample without requiring a high degree of precision or accuracy by the operator or end-user in applying the fluid to the platform, and is advantageous in embodiments of the microsystems platforms of the invention that are used by consumers and other relatively unsophisticated users.
  • the invention also provides laminar flow- dependent replacement of a fluid in a first chamber by a second displacement fluid in a second chamber on the platform.
  • These embodiments of the invention provide approximately complete replacement of a fluid in one chamber on the platform with fluid from another, and thereby provide means for practicing sequential chemical reactions and other sequential processes on the platform under conditions wherein mixing of the two fluids is disadvantageous.
  • the invention also provides turbulent flow mixing components, which permit thorough mixing of different fluid components on the platform.
  • the invention provides mixing chambers fluidly connected with fluid reservoirs containing equal amounts of two or more different fluids or unequal amounts of two or more different fluids.
  • the invention provides fluid reservoirs fluidly connected with mixing chamber of the invention and shaped to determine the relative rate of flow of each of the different fluids into the mixing chamber.
  • gradients of two fluids differing in viscosity, solute concentration or concentration of suspended particulates can be produced using the mixing chambers of the invention.
  • Such gradients can be transferred to reservoirs on the platform for further analytical manipulations, and can form the basis for controlled testing of concentration-dependent effects of various catalysts, drugs, toxins or other biological or chemical agents.
  • Platforms of the invention such as disks and the components comprising such platforms are advantageously provided having a variety of composition and surface coatings appropriate for a particular application.
  • Platform composition will be a function of structural requirements, manufacturing processes, and reagent compatibility/chemical resistance properties.
  • platforms are provided that are made from inorganic crystalline or amorphous materials, e.g.
  • PMMA poly(methyl methacrylate)
  • ABS acetonitrile-butadiene-styrene
  • polycarbonate polyethylene
  • polystyrene polyolefms
  • polypropylene polypropylene and metallocene.
  • Microplatform disks of the invention are preferably fabricated from thermoplastics such as teflon, polyethylene, polypropylene, methylmethacrylates and polycarbonates, among others, due to their ease of molding, stamping and milling.
  • the disks can be made of silica, glass, quartz or inert metal.
  • a fluid handling system is built by sequential application of one or more of these materials laid down in stepwise fashion onto the thermoplastic substrate.
  • Disks of the invention are fabricated with an injection molded, optically-clear base layer having optical pits in the manner of a conventional compact disk (CD).
  • the disk is a round, polycarbonate disk 120mm in diameter and 100 pm thick.
  • the optical pits provide means for encoding instrument control programming, user interface information, graphics and sound specific to the application and driver configuration.
  • the driver configuration depends on whether the micromanipulation device is a handheld, benchtop or floor model, and also on the details of external communication and other specifics of the hardware configuration.
  • This layer is then overlaid with a reflective surface, with appropriate windows for external detectors, specifically optical detectors, being left clear on the disk.
  • Control devices such as integrated circuits, laser diodes, photodiodes and resistive networks that can form selective heating areas or flexible logic structures can be incorporated into appropriately wired recesses, either by direct fabrication of modular installation onto the disk.
  • Reagents that can be stored dry can be introduced into appropriate open chambers by spraying into reservoirs using means similar to inkjet printing heads, and then dried on the disk.
  • a top layer comprising access ports and air vents, ports or shafts is then applied.
  • Liquid reagents are then injected into the appropriate reservoirs, followed by application of a protective cover layer comprising a thin plastic film.
  • the platforms of the invention are preferably provided with a multiplicity of components, either fabricated directly onto the platform, or placed on the platform as prefabricated modules.
  • certain devices and elements can be located external to the platform, optimally positioned on a device of the invention in relation to the platform, or placed in contact with the platform either while rotating or when at rest.
  • Components optimally comprising the platforms of the invention or a controlling device in combination therewith include detection chambers, reservoirs, valving mechanisms, detectors, sensors, temperature control elements, filters, mixing elements, and control systems.
  • This invention provides microsystems platforms comprising the following components.
  • the platforms of the invention are provided comprising microfluidics handling structures in fluidic contract with one another.
  • fluidic contact is provided by capillary or microchannels comprising the surface of the platforms of the invention.
  • MicroChannel sizes are optimally determined by specific applications and by the amount of delivery rates required for each particular embodiment of the platforms and methods of the invention. MicroChannel sizes can range from 0.1 m to a value close to the 1mm thickness of the platform. MicroChannel shapes can be trapezoid, circular or other geometric shapes as required.
  • Microchannels preferably are embedded in a platform having a thickness of about 0.1 to 100mm, wherein the cross-sectional dimension of the microchannels across the thickness dimension of the platform is less than 500 ⁇ m and from 1 to 90 percent of said cross-sectional dimension of the platform.
  • fluid flow is dependent on the orientation of the surfaces of the components. Fluids which completely or partially wet the material of the microchannels, reservoirs, detection chambers, etc. (i.e., the components) of the platforms of the invention which contain them experience a resistance to flow when moving from a component of narrow cross-section to one of larger cross-section, while those fluids which do not wet these materials resist flowing from components of the platforms of the invention of large cross-section to those with smaller cross-section.
  • This capillary pressure varies inversely with the sizes of the two components, or combinations thereof, the surface tension of the fluid, and the contact angle of the fluid on the material of the components .
  • cross-sectional shape Generally, the details of the cross- sectional shape are not important, but the dependence on cross-sectional dimension results in microchannels of dimension less than 500 ⁇ m exhibit significant capillary pressure.
  • "valves" are fashioned that require the application of a particular pressure on the fluid to induce fluid flow. This pressure is applied in the disks of the invention by rotation of the disk (which has been shown above to vary with the square of the rotational frequency, with the radial position and with the extent of the fluid in the radial direction).
  • capillary valves are formed to release fluid flow in a rotation-dependent manner, using rotation rates of from lOOrpm to several thousand rpm. This arrangement allows complex, multistep fluid processes to be carried out using a pre-determined, monotonic increase in rotational rate.
  • FIG. 1 A first example of the microfluidics arrays provided by this invention is shown in Figures 1 through 3.
  • a microsystems platform is provided by the invention that is specifically designed for performing two-step assays. These Figures illustrate an array advantageously used for any two-step analytic process; an antibiotic detection assay is illustrated herein.
  • Such a microsystem platform is illustrated in Figure 1.
  • the arrangement of one assay array 12 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
  • FIG. 2 The components of the antibiotic assay array are shown in greater detail in Figure 2. It will be understood by a comparison of Figures 1 and 2 that the center of the platform 11 is at the top of Figure 2, and the edge or lateral extent of the platform is at the bottom of Figure 2, illustrated by a curved line.
  • Rotation of the antibiotic array on platform disks of the invention can be in either direction, although rotation in a consistent, particular direction is preferred.
  • Disc embodiments of the platforms of the invention were fashioned from machined acrylic and injection-molded polycarbonate.
  • the overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device.
  • the thickness of the disc ranged from about 0.9mm to about 1.5mm.
  • the working fluid volume for reaction with reagents was about l-150 ⁇ L.
  • An entry port 201 having a depth in the platform surface ranging from about 0.25mm to about 6mm and lateral dimensions of from about 0.2cm to about 2cm is constructed on the platform, and designed to accommodate a volume of about 5-100 ⁇ L.
  • This entry port is fluidly connected with an array of metering capillaries 202 having a square cross-sectional diameter ranging from about 0.02mm to about 1mm and proximal ends rounded with respect to entry port 201; the length of this metering capillary array was sufficient to contain a total volume of about 10-5 O ⁇ L.
  • the entry port is also constructed to be fluidly connected with an overflow capillary 203 having a cross-sectional diameter of from about 0.02mm to about 1mm and proximal ends rounded with respect to entry port 201.
  • the overflow capillary is fluidly connected with an overflow chamber 205 having a depth in the platform surface ranging from about 0.02mm to about 5mm, provided that the depth of the overflow chamber 205 is greater than the depth of the overflow capillary
  • Metering capillary 202 is fluidly connected to fluid chamber 204 having a depth in the platform surface ranging from about 0.02mm to about 1mm, provided that the depth of the fluid chamber 204 is greater than the depth of the metering capillary 202.
  • Each of the overflow and fluid chambers is also connected with airports or air channels, such as 211 , that have dimensions ranging from about 0.02mm to about 1mm and permit venting of air displaced by fluid movement on the platform.
  • a capillary junction 212 that is about 0.75mm to 1mm deep is present in the air channel to prevent fluid flow into the air channel.
  • a "capillary junction" is defined in a hydrophilic support material as a pocket, depression or chamber that has a greater depth (vertically within the platform) and/ or a greater width (horizontally within the platform) that the fluid channel to which it is fluidly connected.
  • a contact angle less than 90° such as aqueous solutions on platforms made with most plastics, glass and silica
  • flow is impeded as the channel cross-section increases at the interface of the capillary junction.
  • the force hindering flow is produced by capillary pressure, which is inversely proportional to the cross sectional dimensions of the channel and directly proportional to the surface tension of the liquid, multiplied by the cosine of the contact angle of the fluid in contact with the material comprising the channel.
  • Entry port 201 is positioned on the platform from 1cm to 20cm from the center of rotation.
  • Metering capillary 202 extends from entry port 201 from about 0.2cm to about 20cm.
  • the extent of the length of overflow capillary 203 is at least about 20% greater than the extent of the length of metering capillary 202.
  • the position of fluid chamber 204 is from about 0.5cm to about 10cm from the center of rotation, and the position of overflow chamber 205 is thus from about 1.5cm to about 11.5cm from the axis of rotation.
  • the fluid chamber 204 acts as a capillary barrier that prevents fluid flow from metering capillary 202 at a first, non-zero rotational speed f, sufficient to permit fluid flow comprising overflow from the entry port 201 through overflow capillary 203 and into overflow chamber 205.
  • the capillary boundary of fluid chamber 204 is constructed to be overcome at a second rotational speed f 2 (where f 2 > f,).
  • Fluid chamber 204 is fluidly connected to capillary 206 that is from about 0.02mm to about 1mm deep and has a cross-sectional diameter ranging from about 0.02mm to about 1mm.
  • Capillary 206 extends from about 0.1cm to about 20cm from fluid chamber 204 and is connected to holding chamber 207.
  • Holding chamber 207 has a depth in the platform surface from about 0.02mm to about 1mm and greater than the depth of capillary 206. Filling of fluid chamber 204 is motivated by centripetal acceleration and is accompanied by fluid flow through capillary 206 into holding chamber 207. Holding chamber 207 is fluidly connected by way of capillary 208, having a cross-sectional diameter of from about
  • Read chamber 210 has a depth in the platform surface of from about 0.02mm to about 5mm.
  • read chamber 210 is of sufficient optical quality to prevent excessive scattering for a diffusely-reflecting medium such as milk.
  • read chamber 210 comprises a reflector, for example. This may be a diffuse reflector on the side of the chamber opposite to that upon which light is incident or a mirror- like surface.
  • a diffuse reflector such as paint, a porous material, or any other material which causes diffuse reflection because of its rough surface or porous nature
  • light incident at a given angle is emitted over a hemisphere with a well-known cosine dependence on angle.
  • the detector may be aligned at an angle not equal to the incident angle, so that specular reflection from the plastic window of the chamber does not enter the detector.
  • the mirror-like surface reflects light at an angle equal to the incident angle.
  • a sacrificial valve 213 (as described herein below) is placed as shown in the channel 209. The use of a platform as disclosed in Figures 1 and 2 is illustrated in Figures 3 A through 3J.
  • an imprecise volume (ranging from l-150 ⁇ L of fluid) of a fluid is applied to the entry port 201 (Figure 3 A).
  • the fluid will wick into air channel 211 and be stopped by capillary junction 212.
  • Fluid also wicks into metering capillary 202 and overflow capillary 203.
  • Fluid flows through the metering capillary 202 and overflow capillary 203 at zero rotational speed until the fluid reaches capillary junctions at the junction between metering capillary 202 and fluid chamber 204 and overflow capillary 203 and overflow chamber 205 ( Figures 3B and 3C).
  • Metering capillary 202 is constructed to define a precise volume from about l-150 ⁇ L of fluid between entry port 201 and the capillary junction at fluid chamber 204, which is designed to be at least the amount of the fluid placed by the user in entry port 201.
  • the platform After sample loading by a user and filling of metering capillary 202 and overflow capillary 203 at zero rotational speed (which can be performed on the disc separately or with then disc engaged with a spindle of a centrifugal device), the platform is spun at a first rotational speed f,, ranging from 100-400rpm; the exact value is dependent on the position of the capillary junction components on the platform.
  • this first rotational speed f is equal to about 175rpm for either water or milk.
  • overflow capillary 203 Due to the greater distance of the end of overflow capillary 203 from the center of rotation than the end of metering capillary 202, fluid flows through overflow capillary 203 into overflow chamber 205 and does not flow from metering capillary 202 into fluid chamber 204 at rotational speed ⁇ ,.
  • the platform is spun until all excess fluid is evacuated from entry port 201 and into overflow chamber 205, except the fluid contained in metering capillary 202 ( Figure 3D).
  • a second rotational speed f 2 that is greater than the first rotational speed f l5 typically in the range of 400-520rpm
  • the precise amount of fluid contained in metering capillary 202 is delivered into fluid chamber 204 ( Figures 3E through 3H)
  • this second rotational speed is equal to 400rpm for either water or milk.
  • Fluid movement into fluid chamber 204 is accompanied by filling of capillary 206 and holding chamber 207.
  • sacrificial valves as described above are preferably made of a fungible material that can be removed from the fluid flow path.
  • said sacrificial valves are wax valves and are removed from the fluid flow path by heating, using any of a variety of heating means including infrared illumination and most preferably by activation of heating elements on or embedded in the platform surface as described below.
  • fluid flow is achieved at rotational speed f 2 with removal of the sacrificial valve.
  • capillary 208 preferably fills along with filling of holding chamber 207 until the fluid reaches a capillary junction 209 at the junction between capillary 208 and read chamber 210; in such embodiments, the capillary junction has a depth of from about 0.15mm to about lmm.
  • a third rotational speed f 3 that is greater than the second rotational speed f 2 , typically in the range of >520rpm, the fluid contained in holding chamber 207 is delivered into read chamber 210 ( Figures 31 and 3J).
  • this third rotational speed is equal to 500-800rpm for either water or milk. More generally, the third rotational speed is proportional to l/ (R 0Uter - R ⁇ nner ) x
  • antibiotic detection assay An example of the chemistries of the types of two-step chemical analyses that can be performed using this microfluidics array are illustrated by the antibiotic detection assay.
  • This antibiotic detection assay is performed using the platform design as follows. This chemistry is based on the selective poisoning of an enzyme, carboxypeptidase, by beta lactam antibiotics. Carboxypeptidase is present in a dried form with sugars , buffers or other stabilizers in the holding chamber 207 and a metered amount of milk or other fluid sample solution is introduced into the holding chamber as described above. The milk or other fluid sample solubilizes the enzyme, and the milk/enzyme mixture is incubated at 47° for 3-5 minutes to allow any beta lactam antibiotics present to bind to the carboxypeptidase. The enzyme catalyses cleavage of a D-alanine residue from L-
  • any analytic protocol using the microfluidics structures disclosed herein that requires elevated temperatures advantageously includes resistive heaters as disclosed herein, or other heating elements that are specifically encompassed by the platforms of the invention.
  • the holding chamber 207 contains dried reagent comprising a peptide having a D-amino acid at its carboxyl terminus; an example of such a peptide is L- Lysine-D-Ala-D-Ala.
  • this peptide is also contained in holding chamber 207 in dried form, and is reconstituted by introduction of milk or other fluid into the chamber as described above. It is an advantage of the microfluidics arrays of the invention that a precisely metered amount of fluid is introduced into holding chamber 207, permitting a standardized amount of carboxypeptidase, peptide substrate and buffers, stabilizers, etc. be contained in the holding chamber.
  • D-amino acid oxidase DAAO
  • flavine adenine dinucleotide FAD
  • horse radish peroxidase HRP
  • a chromogen such as syringalazine (4-hydroxy-3,5-dimethoxygenzaldehyde azine) or ortho-dianisidine (ODA) is contained in dried form in read chamber 210.
  • Displacement of the fluid sample from holding chamber 207 to read chamber 210 results in reconstitution of these reagents, which produce a colored product proportional with the amount of D-amino acid present in the fluid sample after incubation with carboxypeptidase and D-amino acid containing peptide.
  • the D-alanine residue produced by degradation of the peptide substrate by carboxypeptidase in holding chamber 207 is degraded into pyruvate in the presence of
  • FADH 2 combines with oxygen in the fluid sample to produce hydrogen peroxide and be reoxidized to FADH 2 to FAD.
  • the hydrogen peroxide then acts on a chromogen such as syringalazine in the presence of horse radish peroxidase, and the previously transparent chromogen becomes colored.
  • chromogen production is detected in the read chamber, and related to the presence of antibiotics in the sample by comparison with samples tested in the absence of antibiotic. Most preferably, a standard curve relating the decrease in chromogen production and the amount of antibiotic in the sample is prepared and used to determine the amount of antibiotic in an unknown test sample.
  • the buffers and reagents used for these chemistries on the platforms of the invention are constituted with appropriate buffers, stabilizers, preservatives, salts, cofactors, adjuvants and other necessary components of the reactions performed on the platform.
  • An alternative embodiment of the two-step assay microfluidics array is shown in Figures 4 through 6, again exemplified for an antibiotic assay disk of the invention. It will be understood that, as in Example 1 , in Figure 4, the arrangement of one assay array 13 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
  • Disk embodiments of the platforms of the invention were fashioned from machined acrylic.
  • the overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device.
  • the thickness of the disc ranged from about 0.9mm to about 1.5mm.
  • the working fluid volume for reaction with reagents was about 25-150 ⁇ L.
  • An entry port 301 having a depth in the platform surface of from about 0.25mm to about 5mm and lateral dimensions of from about 0.2cm to about 2cm is constructed on the platform, and designed to accommodate a volume of about l-100 ⁇ L.
  • This entry port is fluidly connected with one or a multiplicity of entry capillaries 302 having a square cross- sectional diameter ranging from about 0.02mm to about 1mm and having a depth of 0.1 to 1mm equal to the depth of the entry port, and proximal ends rounded with respect to entry port 301.
  • the length of this metering capillary array was sufficient to contain a total volume of about 20 ⁇ L .
  • the entry capillaries 302 are fluidly connected to fluid chamber 303 having a depth in the platform surface of from about 0.02mm to about lmm, wherein the depth is greater than the depth of the entry capillary 302.
  • Each of the fluid chambers of this aspect of the invention is also connected with air ports or air channels, such as 311, that have dimensions ranging from about 0.02mm to about 5mm that permit venting of air displaced by fluid movement on the platform.
  • a capillary junction 312 that is about 0.75mm deep is present in the air channel to prevent fluid flow into the air channel.
  • the fluid chamber 303 is also constructed to be fluidly connected with an overflow capillary 304 having a cross-sectional diameter of about 0.02mm to about 0.75mm and proximal ends rounded with respect to fluid chamber 304.
  • the overflow capillary is fluidly connected with an overflow chamber 306 having a depth in the platform surface of from about 0.02mm to about lmm, greater than the depth of the overflow capillary 304.
  • Entry port 301 is positioned on the platform from 1cm to 20cm from the center of rotation. Entry capillaries 302 extends from entry port 301 from about 0.5cm to about 10cm. The position of a first fluid chamber 303 is from about 0.5cm to about 10cm from the center of rotation.
  • the first fluid chamber 303 acts as a capillary barrier that prevents fluid flow from entry capillary 302 at zero rotational speed. Movement of fluid from entry port 301 through entry capillaries 302 and into the first fluid chamber 303 is achieved by rotation at a first, non-zero rotational speed f , . Displacement of fluid into the first fluid chamber
  • First fluid chamber 303 is also fluidly connected to overflow capillary 304 that is from about 0.05mm to about lmm deep and has a cross-sectional diameter of from about 0.05mm to about lmm and extends from about 0.2cm to about 20cm.
  • Overflow capillary 304 is connected to overflow chamber 306 that has a depth in the platform surface equal to that of overflow capillary 304, so that there is no capillary boundary between overflow capillary 304 and overflow chamber 306.
  • Overflow capillary 304 is positioned in the first fluid chamber 303 at a point radially less distant from entry port 301 than channel 305, thereby defining a volume in the fluid chamber between the position of the overflow capillary 304 and the most radially distant extent of the said first fluid chamber.
  • Second fluid chamber 307 is further fluidly connected through channel 308 to a small pocket or capillary junction 309, having a depth in the platform surface of from about 0.1mm to about 5mm and positioned about 0.2 to 20cm from the axis of rotation.
  • Channel 308 has a cross-sectional diameter ranging from about 0.02mm to about lmm and extends from about 0.2cm to about 10cm, and further extends to a third fluid chamber 310.
  • Third fluid chamber 310 has a depth in the platform surface of from about 0.1mm to about 5mm, that is greater than the depth of capillary 308.
  • Air recirculation channels 311 that have dimensions of from about 0.02mm to about lmm provide pathways for air displaced by fluid movement, while capillary junctions 312 that are about 0.75mm deep prevent fluid from entering the air channels.
  • a sacrificial valve 313 is placed as shown in the channel 309.
  • a valve 314 is placed in channel 305 to control fluid movement from the first fluid chamber 303 to the second fluid chamber 307.
  • FIG. 6A through 6J An imprecise volume (ranging from l-150 ⁇ L of fluid) of a fluid is applied to the entry port 301 ( Figure 6A). Fluid wicks into entry capillary 302 and stops at the capillary junction between entry capillary 302 and the first fluid chamber 303 ( Figures 6B and 6C). Fluid flows through the entry capillary B and into the first fluid chamber 303 at a first rotational speed f l5 ranging from 100-500rpm; the exact value is dependent on the position of the components on the platform ( Figures 6D and 6E). For example, for an entry port 301 having a depth of 0.75mm, entry capillary 302 having dimensions of
  • this first rotational speed f is equal to about 250rpm for either water or milk.
  • the fluid further enters capillary channel 305, stopping at the capillary junction with the second fluid chamber 307. As rotation continues, the fluid continues to fill the first fluid chamber 303, overflow capillary 304 fills (Figure 6F), and excess fluid fills overflow chamber 306 until the level of fluid in the first fluid chamber 303 falls below the position of overflow capillary 304 ( Figure 6G).
  • this second rotational speed is equal to 280rpm for either water or milk.
  • a sacrificial valve 314 is placed at the junction of channel 305 and the second fluid chamber 308, which sacrificial valve is released to permit fluid flow through channel 305 and into the second fluid chamber 308.
  • fluid flow can be achieved at either f, or f 2 rotational velocity.
  • Sacrificial valves are as described above are preferably made of a fungible material that can be removed from the fluid flow path.
  • said sacrificial valves are wax valves and are removed from the fluid flow path by heating, using any of a variety of heating means including infrared illumination and most preferably by activation of heating elements on or embedded in the platform surface as described below.
  • fluid flow is achieved at rotational speed f 2 with removal of the sacrificial valve.
  • capillary 309 preferably fills along with filling of the second chamber 308 until the fluid reaches a capillary junction 309 at the junction between capillary 308 and the third fluid chamber
  • the capillary junction has a depth of about 0.75mm (ranging from about 0.25mm to about lmm).
  • a third rotational speed f that is greater than the second rotational speed f 2 , typically in the range of >500rpm, the fluid contained in the second chamber 308 is delivered into the third fluid chamber 310 ( Figures 6H through 6K).
  • capillary 309 having dimensions of 0.25mm x 0.25mm in cross- section and a length of 3.36-3.7cm from the center of rotation, this third rotational speed is equal to 400rpm for either water or milk.
  • the second fluid chamber 308 contains a reagent, such as carboxypeptidase, and its substrate, for example L-lysine-D-alanine-D-alanine, and incubation is performed therein to produce D-Ala.
  • the remaining reagents are placed in the third fluid chamber 310, and the reaction including color development advantageously proceeds in situ whereby chamber 310 is a read chamber.
  • the extent of chromogen production is detected in the read chamber, and related to the presence of antibiotics in the sample by comparison with samples tested in the absence of antibiotic. Most preferably, a standard curve relating the decrease in chromogen production and the amount of antibiotic in the sample is prepared and used to determine the amount of antibiotic in an unknown test sample.
  • the invention also provides microfluidics arrays for separating the fluid component from a particulate suspension.
  • An example of such a parti culate suspension is blood, where red and white blood cells are suspended in plasma.
  • this aspect of the microfluidics embodiments of the invention is illustrated using separation of blood plasma from whole blood.
  • FIG. 7 A microsystems platform provided by the invention and specifically designed for separating vertebrate blood cells and components is illustrated in Figures 7 through 9.
  • Figure 7 the arrangement of one assay array 14 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
  • FIG. 8 The components of the blood separation array are shown in greater detail in Figure 8. It will be understood by a comparison of Figures 7 and 8 that the center of the platform 11 is at the top of Figure 8, and the edge or lateral extent of the platform is at the bottom of Figure 8, illustrated by a curved line.
  • Rotation of the blood separation array on platform disks of the invention can be in either direction, although rotation in a consistent, particular direction is preferred.
  • Disk embodiments of the platforms of the invention were fashioned from machined acrylic.
  • the overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device.
  • the thickness of the disc ranged from about 0.9mm to about 1.5mm.
  • the working fluid volume was about l-50 ⁇ L.
  • An entry port 401 having a depth in the platform surface ranging from about 0.1mm to about 5mm and lateral dimensions of about 0.1 to about 2cm is constructed on the platform, and designed to accommodate a volume of about 5 to about 50 ⁇ L.
  • This entry port is fluidly connected to an entry capillary 402, having a cross-sectional diameter of from about 0.02mm to about lmm and a depth of about 0.5 to lmm.
  • the length of this entry capillary was sufficient to contain a total volume of from about 1 to about 15 ⁇ L.
  • Entry capillary 402 is further fluidly connected to a separation column 403 having a cross- sectional diameter of from about 0.1mm to about 2mm, a depth of from about 0.25mm to about lmm, and a length sufficient to contain a total volume of 10 to about 20 ⁇ L.
  • This separation column is also fluidly connected with a passage 411 to overflow chamber 404.
  • Passage 411 has a cross-sectional diameter ranging from about 0.5mm to about 2mm, a depth of about 0.25mm to about lmm, and a length of 0.5mm to about 5mm).
  • Overflow chamber 404 has a depth of from about 0.25-lmm.
  • a small capillary exit 406 is also fluidly connected with separation chamber 403, having a cross-sectional diameter of from about 0.05mm to about0.25mm, a depth of about 0.025mm to about 0.125mm, and a length about 0.25mm to about 5mm.
  • This capillary is arranged to traverse a direction radially more proximal to the axis of rotation than the insertion point with separation column 403.
  • This small capillary 406 terminates in a capillary junction 407 that is fluidly connected with capillary 408, extending in a radial direction to decant chamber 405.
  • a sacrificial valve 413 is positioned in small capillary 406 at the juncture with the capillary junction.
  • Capillary 408 has a cross- sectional diameter ranging from about 0.05mm to about lmm, a depth of about 0.05mm to about lmm, and a length of from about lmm to about 100mm.
  • This capillary is arranged in a radially outward direction between capillary junction 407 and decant chamber 405.
  • Passage 411 is positioned on separation column 403 to be significantly more proximal to the axis of rotation than the insertion point of small capillary 406.
  • Air displacement channels 409 that have dimensions of from about 0.02mm to about lmm permit venting of air displaced by fluid movement on the platform.
  • Capillary junctions 410 that are about 0.75mm deep are present in the air channels to prevent fluid flow into the air channels.
  • FIGs 9A through 9H The use of this platform is illustrated in Figures 9A through 9H for separating plasma from whole blood.
  • An imprecise volume (ranging from l-150 ⁇ L of fluid) of blood is applied to the entry port 401 ( Figure 9A).
  • Blood enters the entry capillary 402 by capillary action, and stops at the capillary junction between entry capillary 402 and the separation chamber 403 ( Figures 9B and 9C).
  • a first rotational speed f ranging from 100-300rpm (the exact value is dependent on the position of the components on the platform), blood flows from the entry capillary 402 into separation chamber 403 ( Figure 9D).
  • small channel 406 has dimensions that prevent wicking of blood into the channel as blood flows past the insertion point of small channel 406 into separation column 403.
  • Fluid flow of plasma into decant chamber 405 results from fluid flow overcoming the capillary barrier 407 by rotation at a third rotational speed f 3 , that is greater than the second rotational speed f 2 , typically in the range of >1000-5000rpm (Figure 9H).
  • An alternative embodiment of the fluid separation platform is also provided by the invention, again illustrated by the separation of plasma from whole blood.
  • This embodiment of the blood separation microfluidics array is shown in Figures 10 through 12. It will be understood that, as above, in Figure 10, the arrangement of one separation array 15 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
  • Disk embodiments of the platforms of the invention are fashioned from machined acrylic.
  • the overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device.
  • the thickness of the disc ranged from about 0.9mm to about 1.5mm.
  • the working fluid volume was about 5-50 ⁇ L.
  • An entry port 501 having a depth in the platform surface of about 0.1mm to about lmm and lateral dimensions of about 0.1cm to about 2cm is constructed on the platform, and designed to accommodate a volume of about 5 to about 50 ⁇ L.
  • This entry port is fluidly connected with a first array of metering capillaries 502 and a second array of metering capillaries
  • each of the capillaries has a cross-sectional diameter of from about 0.02mm to about lmm.
  • the length of the second metering capillary array 503 is longer than that of the first metering capillary array 502.
  • the first metering capillary array 502 is fluidly connected with a ballast chamber 507, having a depth in the platform surface ranging radially from about 0.1mm to about 5mm and greater than the depth of the first metering capillary array 502, wherein the first metering capillary array 502 forms a capillary junction between the array and the ballast chamber.
  • the second capillary array 503 is fluidly connected with capillary junction 506.
  • the entry port is also constructed to be fluidly connected with an overflow capillary 504 having a cross-sectional diameter of about 0.02mm to about lmm and proximal ends rounded with respect to entry port 501.
  • the overflow capillary is fluidly connected with an overflow chamber 505 having a depth in the platform surface of from about 0. lmm to about 5mm, greater than the depth of the overflow capillary 504.
  • Each of the overflow and fluid chambers is also connected with airports or air channels, such as 514, that have dimensions ranging from about 0.1mm to about lmm and that permit venting of air displaced by fluid movement on the platform.
  • Capillary junctions 516 that are about 0.75mm deep are present in the air channels to prevent fluid flow into the air channels.
  • Entry port 501 is positioned on the platform from 0.5cm to 20cm from the center of rotation.
  • Metering capillary array 502 extends from about 0.6cm to about 10cm from entry port 501.
  • Metering capillary array 503 extends about 0.5cm to about 10cm from entry port 501.
  • the length of metering capillary array 503 is about at least about 20% longer than metering capillary array 502, and the extent of the length of overflow capillary 504 is at least about 20% greater than the extent of the length of either the first metering capillary array 502 or the second metering capillary array 503.
  • ballast chamber 507 is about 1cm to about 10cm from the center of rotation
  • the position of capillary junction 506 is about 1.5 to 15cm from the center of rotation
  • the position of overflow chamber 505 is thus about 2.5 to about 20cm from the axis of rotation.
  • the ballast chamber 507 acts as a capillary barrier that prevents fluid flow from the first metering capillary array 502 at a first, non-zero rotational speed fj sufficient to permit fluid flow comprising excess blood overflow from the entry port 501 through overflow capillary 504 and into overflow chamber 505.
  • Capillary junction 506 is a capillary barrier that prevents fluid flow from the second metering capillary array 503 at said first, non-zero rotational speed f, sufficient to permit fluid flow comprising excess blood overflow from the entry port 501 through overflow capillary 504 and into overflow chamber 505.
  • These capillary boundaries are constructed to be overcome at a second rotational speed f 2 (where f 2 > f,).
  • Ballast chamber 508 is fluidly connected to capillary 510 that is from about 0.02mm to about lmm deep and has a cross-sectional diameter of about 0.02mm to about lmm and that extends from about 0.1cm to about 5cm.
  • Capillary 510 is connected to capillary junction 511.
  • capillary 510 is fluidly connected with a sacrificial valve 515.
  • Sacrificial valves as described below are preferably made of a fungible material that can be removed from the fluid flow path.
  • said sacrificial valves are wax valves and are removed from the fluid flow path by heating, using any of a variety of heating means including infrared illumination and most preferably by activation of heating elements on or embedded in the platform surface.
  • fluid flow is achieved at rotational speed f 2 with removal of the sacrificial valve.
  • Sacrificial valve 515 or capillary junction 511 are further fluidly connected with channel 512 which is from about 0.1mm to about lmm deep and has a cross-sectional diameter of about 0.1mm to about lmm.
  • Channel 512 extends about 0.1cm to about 20cm and is fluidly connected with separation chamber 509 at a point most distal from the axis of rotation.
  • Second metering capillary array 503 is fluidly connected with capillary junction 506, which is overcome at a rotational speed f 2 > f,.
  • Capillary junction 506 is further fluidly connected to channel 508, which is further fluidly connected to separation chamber 509.
  • Channel 508 is from about 0.02mm to about lmm deep and has a cross- sectional diameter of from about 0.02mm to about lmm.
  • Channel 508 extends from about t _cm to about jkcm.
  • Separation chamber 509 is from about 0.2mm to about 5mm deep and has a cross-sectional dimension ranging from about lmm to about 20mm, and is positioned from about 10mm to about 100mm from the center of rotation.
  • Separation chamber 509 is fluidly connected with decant channel 517 at a point close to the chamber's most axis-proximal extent.
  • Decant channel 517 is about 0.02mm to about lmm deep and has a cross-sectional diameter ranging from about 0.02mm to about lmm.
  • Decant channel 517 extends from about 4.3cm to about 5cm and is fluidly connected with decant chamber 514.
  • Decant chamber 514 is from about 0.2mm to about 2mm deep and has a cross-sectional diameter of from about lmm to about 10mm. Decant chamber 514 is positioned about 5.2cm from the center of rotation..
  • FIG. 12A The use of this embodiment of the microfluidics separation arrays of the invention is illustrated in Figures 12A through 12J.
  • An imprecise volume (ranging from l-150 ⁇ L of fluid) of blood is applied to the entry port 501 ( Figure 12A).
  • Blood enters the each of the metering capillary arrays 502 and 503 and stops at the capillary junction between metering capillary array 502 and ballast chamber 507 and between metering capillary 503 and capillary junction 506 ( Figures 12B and 12C). Blood also enters and fills overflow capillary 504, stopping at the capillary junction with overflow chamber 505.
  • first rotational speed f ranging from 100-500rpm (the exact value is dependent on the position of the components on the platform)
  • blood flows from the entry port 501 through overflow capillary 504 and into overflow chamber 505 ( Figures 12D and 12E).
  • second rotational speed f 2 that is greater than the first rotational speed f, typically in the range of 300-800rpm
  • the capillary junction between the first metering capillary array 502 and ballast chamber 508 is overcome, and blood from the first metering capillary array fills ballast chamber 508 ( Figure 12F).
  • the separated plasma does not extend to decant capillary 517.
  • Decant channel 517 is advantageously provided with dimensions that retard passage of unfractionated blood, or plasma contaminated with greater than 0.1-1% of blood cells found in whole blood.
  • the invention also provides mixing chambers and arrays of mixing chambers for mixing two or more fluids that differ in viscosity, solute concentration or concentration of suspended particulates.
  • a first embodiment of microfluidics platform comprising the mixing chambers and arrays of the invention is shown in Figures 13- 15 for mixing equal volumes of different liquids.
  • Figure 13 the arrangement of one assay array 15 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
  • FIG. 11 is at the top of Figure 14, and the edge or lateral extent of the platform is at the bottom of Figure 14, illustrated by a curved line.
  • Rotation of the mixing array on platform disks of the invention can be in either direction, although rotation in a consistent, particular direction is preferred.
  • Disk embodiments of the platforms of the invention were fashioned from machined acrylic.
  • the overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device.
  • the thickness of the disc ranged from about 0.9mm to about 1.5mm.
  • the working fluid volume was about 50 ⁇ L.
  • Entry ports 601 having a depth in the platform surface ranging from about 0.1mm to about lmm and lateral dimensions of from about 1cm to about 5cm are constructed on the platform, and designed to accommodate a volume of about 5-50 ⁇ L.
  • Each entry port is fluidly connected with one of a paired array of metering capillaries 602 having a square cross- sectional diameter of from about 0.1mm to about 0.5mm and proximal ends rounded with respect to entry ports 601 ; the length of each metering capillary array was sufficient to contain a total volume of about 25 ⁇ L.
  • Metering capillaries 602 are fluidly connected to a curved capillary barrier 603 having a depth in the platform surface of about 0.25mm to about lmm that is greater than the depth of metering capillaries 602.
  • the capillary barrier 603 and other fluid components of the mixing array are also connected with air channels 608, that have dimensions ranging from about 0.25mm to about lmm and that permit venting of air displaced by fluid movement on the platform.
  • capillary junctions 609 that are about 0.75mm deep are present in the air channels to prevent fluid backflow into the air channel.
  • Capillary barrier 603 is fluidly connected by a na ⁇ ow capillary channel 604 to mixing chamber 605, which is fluidly connected with channel 610, which is further connected with mixed fluid receiving chamber 606.
  • capillary 604 comprises a sacrificial valve 612. Sacrificial valves used in this embodiment of the invention are as described below.
  • Capillary channel 604 ranges from about 0.1mm to about lmm in depth and has a cross-sectional diameter of from about 0.1mm to about lmm).
  • Capillary channel 604 extends from about 0.1 to about 10 cm.
  • Mixing chamber 605 is about 0. lmm to about lmm deep and has a cross-sectional diameter of from about
  • Capillary channel 610 ranges from about 0.1mm to about lmm deep and has a cross-sectional diameter of from about 0.1 mm to about 1 mm). Capillary channel 610 extends from about 0.2cm to about 30cm.
  • mixing chamber 605 is constructed such that the insertion point of capillary channel 604 and the insertion point of capillary channel 610 are offset at opposite ends of the mixing chamber. As a consequence, fluid flowing through capillary channel 604 is forced to encounter the opposite wall of mixing chamber 605 before fluid flow can proceed through capillary channel 610.
  • the turbulence created by the structure of mixing chamber 605 is sufficient to disrupt laminar flow and cause fluid mixing in the chamber prior to continued fluid flow through capillary channel 610 and into mixed fluid receiving chamber 606.
  • the positions of the capillaries 604 and 610 can be at any convenient position in mixing chamber 605, wherein the Coriolis forces of fluid flow are sufficient to disrupt laminar flow and provide turbulence that results in efficient mixing.
  • Mixed fluid receiving chamber 606 is about 0.1mm to about 5mm deep, has a cross-sectional diameter of from about lmm to about 20mm, and is positioned from about 1cm to about 30cm from the center of rotation.
  • the fluids from each capillary a ⁇ ay flows into and fills the capillary barrier 603 ( Figure 15B).
  • the valve prevents fluid flow into channel 604; otherwise, fluid flow proceeds into channel 604 at rotational speed f 1#
  • fluid flow proceeds from capillary junction 603 through channel 604 and into mixing chamber 605 ( Figure 15C).
  • Fluid flow within mixing chamber 605 is turbulent, in contrast to fluid flow through capillary barrier 603 or channel 604, which is primarily laminar, so that mixing occurs predominantly in mixing chamber 605. Fluid flow proceeds through channel 610 and the mixed fluid solution is displaced into mixed fluid receiving chamber 606 ( Figure 15D).
  • the invention also provides mixing arrays for mixing unequal volumes of fluid.
  • FIG. 19 One example of such an additional embodiment of the microsystems platform provided by the invention and specifically designed for performing mixing of unequal volumes of different liquid samples is illustrated in Figures 19 through 21.
  • Figure 19 the arrangement of one assay array 17 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
  • the components of the mixing array are shown in greater detail in Figure 20. It will be understood by a comparison of Figures 19 and 20 that the center of the platform 11 is at the top of Figure 20, and the edge or lateral extent of the platform is at the bottom of Figure 20, illustrated by a curved line.
  • Rotation of the mixing array on platform disks of the invention can be in either direction, although rotation in a consistent, particular direction is preferred.
  • Disk embodiments of the platforms of the invention are fashioned from machined acrylic.
  • the overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device.
  • the thickness of the disc ranged from about 0.9mm to about 1.5mm.
  • the working fluid volume is about 2-200 ⁇ L.
  • Fluid reservoirs 701 and 702 each containing one of a pair of liquids to be mixed, are constructed on the platform, having a depth in the platform surface ranging from about 0.1mm to about 5mm and lateral dimensions of from about 0.2cm to about 10cm).
  • fluid reservoir 701 is designed to accommodate a volume of about 1 to about 500 ⁇ L
  • fluid reservoir 702 is designed to accommodate a volume of about ranging from about 1 to about 500 ⁇ L, wherein the volume of fluid reservoir 702 is less than the volume of fluid reservoir 701.
  • the viscosity of the fluid in the fluid reservoirs may differ, so that mixing produces a mixed fluid of intermediate viscosity.
  • each fluid reservoir is fluidly connected with a capillary channel 703 or 704 to capillary junction 705.
  • Each capillary channel is from about 0.0.2mm to about lmm deep, has a cross-sectional diameter ranging from about 0.1mm to about lmm, and extends about 2cm to about 100cm.
  • Capillary junction 705 has a depth in the platform surface ranging from about 0.02mm to about 1 mm that is greater than the depth of capillaries 703 to 704.
  • capillaries 703 or 704 comprise a sacrificial valve 712. Sacrificial valves used in this embodiment of the invention are as described below. Use of said sacrificial valves can be used in addition to or in place of capillary junction 705.
  • the fluid components of the mixing array are also connected with air channels
  • capillary junctions 711 that are about 0.75mm deep are present in the air channels to prevent fluid backflow into the air channel.
  • Capillary junction 705 is fluidly connected by a narrow capillary channel 706 to mixing chamber 707, which is fluidly connected with channel 708, which is further connected with mixed fluid receiving chamber 709.
  • capillary 706 comprises a sacrificial valve 712, as described below.
  • Capillary channel 706 ranges from about 0.1mm to about lmm, has a cross-sectional diameter of from about 0.1mm to about lmm and extends from about 0.2cm to about 30cm.
  • Mixing chamber 707 is from about 0.1mm to about lmm deep, has a cross-sectional diameter ranging from about 0. lmm to about lmm, and is positioned from about 0.2cm to about 30cm from the center of rotation.
  • Capillary channel 708 ranges from about 0.1mm to about lmm, has a cross-sectional diameter ranging from about lmm to about 20mm and extends from about 0.2cm to about 30cm.
  • Capillary channel 706 and capillary channel 708 are advantageously offset in their connection with the mixing chamber as described above, or are positioned at any convenient position in the mixing chamber for those embodiments relying on Coriolis forces to create mixing.
  • Capillary 708 is fluidly connected with mixed fluid receiving chamber 709.
  • Mixed fluid receiving chamber 709 is about 0.1mm to about lmm, has a cross-sectional diameter of about lmm to about 20mm, and is positioned from about 1cm to about 30cm from the center of rotation.
  • FIG. 21A through 21E The use of this embodiment of the microfluidics components of the invention is illustrated in Figures 21A through 21E.
  • a volume (ranging from l-150 ⁇ L of fluid) of each of the fluids to be mixed is applied to the fluid reservoirs 701 and 702 ( Figure 21 A). Fluid enters the each of the capillaries 703 and 704 and stops at capillary junction
  • the platforms of the invention are provided containing the fluids to be mixed already in fluid reservoirs 701 and 702.
  • sacrificial valves 712 be provided in capillaries 703 and 704, to prevent evaporation, wetting or leakage of fluid from the reservoirs prior to use.
  • f first rotational speed
  • the fluids from each capillary flows past capillary junction 605 and through mixing chamber 707 ( Figures 21B and 21C).
  • the valve prevents fluid flow into channels 703 and 704.
  • mixing chamber 707 Upon release of sacrificial valve 712, fluid flow proceeds from capillary junction 705 through channel 706 and into mixing chamber 707 ( Figure 21C). Fluid flow within mixing chamber 707 is turbulent, in contrast to fluid flow through capillary barrier 705 or channel 706, which is primarily laminar, so that mixing occurs predominantly in mixing chamber 707. Fluid flow proceed through channel 708 and the mixed fluid solution is displaced into mixed fluid receiving chamber 709 ( Figures 2 ID and 2 IE).
  • platforms capable of forming a gradient of two or more liquids that differ in viscosity, solute concentration or concentration of suspended particulates.
  • FIGs 22-24 Such an additional embodiment of the microsystems platform provided by the invention and specifically designed for performing mixing of different volumes of liquid samples to form a gradient in the concentration of a species in which the two fluids differ is illustrated in Figures 22-24.
  • Figure 22 the arrangement of one assay array 18 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
  • the components of the mixing array are shown in greater detail in Figure 23. It will be understood by a comparison of Figures 22 and 23 that the center of the platform 11 is at the top of Figure 23, and the edge or lateral extent of the platform is at the bottom of Figure 23, illustrated by a curved line.
  • Rotation of the mixing array on platform disks of the invention can be in either direction, although rotation in a consistent, particular direction is preferred.
  • Disk embodiments of the platforms of the invention are fashioned from machined acrylic.
  • the overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device.
  • the thickness of the disc ranged from about 0.9mm to about 1.5mm.
  • the working fluid volume is about 40 ⁇ L.
  • the components of the mixing array are as follows. Fluid reservoirs 801 and
  • Fluid reservoir 801 is designed to accommodate a volume of from about 1 to about 500 ⁇ L
  • fluid reservoir 802 is designed to accommodate a volume of about 1 to about 500 ⁇ L, wherein the shape of fluid reservoir 802 is different than the shape of fluid reservoir 801.
  • fluid reservoirs 801 and 802 are shaped so that the rate of fluid output in the two reservoirs differs between the reservoirs at a particular rotational speed, due to a change in the pressure "head" (related to the cross-sectional area of the fluid at each point in the reservoir), so that the proportion of fluid in the mixture from one of the reservoirs is at a maximum at the beginning of rotation and is at a minimum when the fluids from the reservoirs are completely mixed at the end of rotation, thus forming a gradient.
  • Gradients produced according to this aspect of the invention can consist of salt gradients, including sodium chloride and cesium chloride or sulfate gradients, gradients of low molecular weight species such as sucrose, synthetic polymer gradients such as Ficoll or Hypaque, or gradients of a drug, toxin, enzyme substrate or other small molecule of interest.
  • Each fluid reservoir is fluidly connected with a capillary channel 803 or 804 to capillary junction 805.
  • Each capillary channel ranges from about 0.1mm to about lmm deep, has a cross-sectional diameter of from about 0.1mm to about lmm and extends about 2cm to about 100cm.
  • Capillary junction 805 has a depth in the platform surface of from about 0.1mm to about lmm that is greater than the depth of capillaries 803 to 804.
  • capillaries 803 or 804 comprise a sacrificial valve 812, as described below. Use of said sacrificial valves can be used in addition to or in place of capillary junction 805.
  • the fluid components of the mixing array are also connected with air channels
  • capillary junction 810 that have dimensions of from about 0. lmm to about lmm and permit venting of air displaced by fluid movement on the platform.
  • capillary junctions 811 that are about 0.75mm deep are present in the air channels to prevent fluid backflow into the air channel.
  • Capillary junction 805 is fluidly connected by a narrow capillary channel 806 to mixing chamber 807, which is fluidly connected with channel 808, which is further connected with mixed fluid receiving chamber 809.
  • capillary 806 comprises a sacrificial valve 812.
  • Capillary channel 806 is from about 0.1mm to about lmm, has a cross-sectional diameter of from about 0.1mm to about lmm and extends from about 0.2cm to about 30cm.
  • Mixing chamber 807 is from about 0.1mm to about lmm, has a cross-sectional diameter of from about 0.1mm to about lmm, and is positioned from about 0.2cm to about 30cm from the center of rotation.
  • Capillary channel 808 is from about 0. lmm to about lmm, has a cross-sectional diameter of about lmm to about 20mm and extends from about 0.2cm to about 30cm.
  • Capillary channel 806 and capillary channel 808 are advantageously offset in their connection with the mixing chamber as described above, or are positioned at any convenient position in the mixing chamber for those embodiments relying on Coriolis forces to create mixing.
  • Capillary 808 is fluidly connected with mixed fluid receiving chamber 809.
  • Mixed fluid receiving chamber 809 is about 0.75mm deep (ranging from about 0.1mm to about lmm) and having a cross-sectional diameter of about 5mm (ranging from about lmm to about 20mm) from about, and is positioned from about 1 cm to about 30cm from the center of rotation.
  • FIGS 24A through 24E Use of this microfluidics platform to produce a gradient as described herein is illustrated in Figures 24A through 24E.
  • a volume (ranging from l-150 ⁇ L of fluid) of each of the fluids to be mixed is applied to the fluid reservoirs 801 and 802 ( Figure 24 A). Fluid enters the each of the capillaries 803 and 804 and stops at capillary junction
  • the platforms of the invention are provided containing the fluids to be mixed already in fluid reservoirs 801 and 802.
  • sacrificial valves 812 be provided in capillaries 803 and 804, to prevent evaporation, wetting or leakage of fluid from the reservoirs prior to use.
  • a first rotational speed f ranging from 100-lOOOrpm (the exact value is dependent on the position of the components on the platform)
  • the fluids from each capillary flows past capillary junction 805 and through mixing chamber 807 ( Figures 24B and 24C).
  • the valve prevents fluid flow into channels 803 and 804.
  • fluid flow proceeds from capillary j unction 805 through channel 806 and into mixing chamber 807
  • FIG. 24D Fluid flow within mixing chamber 807 is turbulent, in contrast to fluid flow through capillary barrier 805 or channel 806, which is primarily laminar, so that mixing occurs predominantly in mixing chamber 807. Fluid flow proceed through channel 808 and the mixed fluid solution is displaced into mixed fluid receiving chamber 809 ( Figures 24D and 24E).
  • the invention provides microfluidics arrays comprising a multiplicity of mixing chambers fluidly connected in series to one another. Such an arrangement is illustrated in Figure 28.
  • mixing cascades as shown in Figure 28 are useful for mixing liquids of dramatically different volumes or viscosities, such as mixing a low volume, high viscosity liquid with a high volume, low viscosity liquid.
  • the mixing array comprises a multiplicity of mixing chambers a ⁇ ayed radially across the platform surface, each mixing chamber having an inlet capillary extending radially from a position more proximal to the center of rotation than the mixing chamber, and an outlet capillary extending radially to a position more distal to the center of rotation than the mixing chamber.
  • the capillaries are connected with the mixing chamber so that their positions in the mixing chamber are offset from one another, wherein fluid flow from the inlet capillary impinges on a wall of the mixing chamber at a position other than the position occupied by the outlet capillary, thereby producing turbulent flow within the mixing chamber that mixes the fluids.
  • the capillaries can be positioned in the mixing chamber at any convenient position, and Coriolis forces have be relied upon to facilitate mixing.
  • the inlet capillary of the first mixing chamber is fluidly connected with the fluid reservoirs (either directly or through a capillary junction), and the inlet capillary of the other mixing chamber in the array is the outlet channel of the mixing chamber immediately more proximal to the center of rotation; similarly, the outlet capillary of each mixing chamber is the inlet capillary of the mixing chamber immediately more distal to the center of rotation, and wherein the outlet capillary of the mixing chamber positioned the most distal from the center of rotation is fluidly connected with a mixed fluid receiving chamber.
  • the fluid is repetitively mixed in a multiplicity of mixing chambers arrayed more and more distally from the center of rotation, ending with a reservoir or chamber sufficient in volume to accommodate the volume of the mixed fluid.
  • the dimensions of the capillaries, fluid reservoirs, mixing chambers and mixed fluid receiving chamber in these arrays are as described above.
  • Figure 28 also illustrates another embodiment of the mixing arrays of the invention.
  • a specialized mixed fluid receiving chamber termed a gradient chamber, is provided; one embodiment of this receiving chamber is shown in
  • This chamber permits the gradient fluid stream to be aliquotted into the individual compartments of the chamber, wherein the concentration of the gradient decreases with increasing distance from the gradient chamber inlet capillary.
  • the chamber can be modified to contain a detection means in each compartment, so that the concentration effect of the changing component of the gradient can be determined.
  • microfluidics platforms of this invention are provided a microsystems platform specifically designed for performing a specific binding assay. These embodiments are exemplified using an immunoassay as illustrated in Figures 25 through 27.
  • Figure 25 the arrangement of one assay array 19 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
  • the components of the mixing array are shown in greater detail in Figure 26.
  • an incubation chamber that comprises a specific binding reagent.
  • specific binding reagent is intended to encompass biomolecules having a specific binding affinity between pairs thereof providing a specific molecular binding interaction with a binding affinity constant of between about 10 ⁇ 4 and 10 15 M.
  • pairs of specific binding reagents include but are not limited to antigen and antibody, including antisera, polyclonal antibodies and most preferably monoclonal antibodies; receptor and ligands, including cell-surface receptors; integrins and adhesion proteins, including ICAM-I and ICAM-II; and carbohydrates and lectins, including phytohemagglutinin.
  • specific binding reagents comprising a first member of a specific binding pair is provided in an incubation chamber as a coating on the surface of the chamber, as a dried or lyophilized reagent contained in the chamber and reconstituted upon addition of a fluid or fluid sample to the chamber; contained on a support media, such as latex or other beads, or in a gel or other support media.
  • Said first member of a specific binding pair is designed or intended to detect the presence of an analyte, for example, a cell expressing a cognate antigen, receptor or adhesion protein or having a carbohydrate moiety at the cell surface specific for a particular lectin.
  • Said specific binding reagent is applied to the incubation chamber of the platform by depositing the reagent on the surface using any appropriate means, including inkjet printing, computer-positioned syringes, microetching and microlithographic methods, including photolithography, screen and airbrush printing methods, solution coating, dipping, and conventional microtitre-well techniques.
  • the surface or detection chamber can be treated to provide a two-dimensional array or pattern, wherein certain areas on the surface or detection chamber are treated with said specific binding reagent and others are not in a recognizable manner.
  • the components of the specific binding assay array are as follows.
  • metering capillary 902 having a depth in the platform surface ranging from about 0.1mm to about lmm deep and lateral dimensions of about 0.1cm to about 2cm is constructed on the platform, and designed to accommodate a volume of about from 2 ⁇ L to lOO ⁇ L.
  • This entry port is fluidly connected with a metering capillary 902 having cross-sectional diameter of from about 0.1mm to about lmm, and having a depth of about 0.25mm to lmm; the length of this metering capillary was sufficient to contain a total volume of about 2 to about 1 OO ⁇ L.
  • the metering capillary 902 is fluidly connected to capillary junction 904.
  • the entry port is also constructed to be fluidly connected with an overflow capillary 903 having a cross-sectional diameter of about 0.1mm to about lmm and proximal ends rounded with respect to entry port 901.
  • the overflow capillary is fluidly connected with an overflow chamber 905 having a depth in the platform surface of about
  • Each of the overflow and fluid chambers is also connected with air ports or air channels, such as 523, that have dimensions of about 0.1mm to about lmm and that permit venting of air displaced by fluid movement on the platform.
  • Capillary junctions 524 that are about 0.75mm deep are present in the air channels to prevent fluid flow into the air channels.
  • Entry port 901 is positioned on the platform from 1cm to 20cm from the center of rotation.
  • Metering capillary 902 extends from entry port 901 from about 0.5cm to about 5cm.
  • Overflow capillary 902 extends from entry port 901 from about 1cm to about 20cm.
  • the extent of the length of overflow capillary 903 is 20% longer than metering capillary 902.
  • the position of overflow chamber 905 is from about 1cm to about 20cm from the center of rotation, and the position of capillary junction 904 is from about 1cm to about 20cm from the center of rotation.
  • Capillary junction 904 is fluidly connected with capillary channel 906, which in turn is fluidly connected with incubation chamber 910.
  • Capillary channel 906 has a cross-sectional diameter of about 0.1mm to about lmm and extends from about 0.2cm to about 10cm from the capillary junction.
  • Incubation chamber 910 has a depth in the platform surface ranging from about 0. lmm to about lmm, that is greater than the depth of capillary channel 906.
  • Capillary channel 906 is also fluidly connected with channel 909 through capillary junction 907.
  • Capillary junction 907 is constructed to prevent fluid flow backwards through the junction.
  • Channel 909 has a cross-sectional diameter of about 0.1mm to about lmm and extends from about 0.2cm to about 5cm from the capillary junction.
  • Capillary junction 907 has a depth in the platform surface of about 0.1 mm to about 1 mm, greater than the depth of the channel 909 or capillary channel 906.
  • Incubation chamber 910 also contains a specific binding species, most preferably an antibody, specific for a component of the sample. This species is advantageously contained within incubation chamber 910 as a coating on the surface of the chamber, or attached to beads or other carrier within the chamber, or to a functionalized inner surface of the chamber or otherwise as described above.
  • Capillary junction 907 is further fluidly connected with wash buffer reservoir 516, having a depth in the platform surface of from about 0.1mm to about lmm and positioned at a distance from about 10mm to about 200mm from the axis of rotation.
  • Capillary junction 907 is further fluidly connected with reagent capillary 920, which is further fluidly connected with capillary junction 914, which is further fluidly connected with channel 926, and which is fluidly connected with reagent reservoir 917.
  • Reagent capillary 920 has a cross-sectional diameter of about 0.1mm to about lmm and extends from about 0.2cm to about 20cm reagent reservoir 917.
  • Capillary junction 914 has a depth in the platform surface ranging from about 0.1mm to about lmm and is positioned at a distance from about 10mm to about 200mm from the axis of rotation.
  • Reagent capillary 926 has a cross-sectional diameter of from about 0.1mm to about lmm and extends from about 0.2cm to about 20cm from the capillary junction.
  • Reagent reservoir 917 has a depth in the platform surface of about 0.1mm to about lmm, and is positioned at a distance from about 10mm to about 200mm from the axis of rotation.
  • Incubation chamber 910 is fluidly connected at a point most distal to the axis of rotation to U-shaped capillary 921.
  • U-shaped capillary 921 has a cross-sectional diameter of about 0.1mm to about lmm and extends from about 0.2cm to about 20cm between incubation chamber 910 and waste reservoir 915 . This capillary extends in a
  • U-shape to a point that is at least as proximal to the axis of rotation than the most axis- proximal extent of incubation chamber 910. This positioning of the U-shaped channel relative to incubation chamber 910 ensures that additional fluids flowing into incubation chamber 910 and displacing fluid therefrom will displace said fluid homogeneously, i. e. , the first fluid in the chamber will be pushed out of the chamber whilst being replaced by the second fluid.
  • Waste reservoir 915 has a depth in the platform surface of about 0.1mm to about 5mm and is positioned at a distance from about 10mm to about 200mm from the axis of rotation; as shown in Figure 26, the waste reservoir is typically positioned at the farthest distance from the axis of rotation of any of the components of the array.
  • sacrificial valves 922 can be positioned at the junction of capillary junction 904 and capillary channel 906, at the junction of capillary junction 907 and wash buffer capillary 908, or at the junction of reagent reservoir 918 and capillary junction 919. The use of this platform for performing an immunoassay is illustrated in Figures
  • reagent reservoir 916 and wash reservoir 915 are pre-loaded on the disk, and most preferably the disk contains sacrificial valves 922 at the junction of capillary junction 907 and wash buffer capillary 908, and at the junction of reagent reservoir 918 and capillary junction 919.
  • An imprecise volume (ranging from 1-15 O ⁇ L of fluid) of a fluid is applied to the entry port 901 ( Figure 27A).
  • Fluid wicks into metering capillary 902 and stops at the capillary junction between metering capillary 902 and capillary junction 904 ( Figures 27B and 27C).
  • the platform is spun at a first rotational speed f,, ranging from 100- 500rpm; the exact value is dependent on the position of the components on the platform.
  • overflow capillary 903 Due to the greater distance of the end of overflow capillary 903 from the center of rotation than the end of metering capillary 902, fluid flows through overflow capillary
  • a second rotational speed f 2 that is greater than the first rotational speed f, typically in the range of 100-1 OOOrpm
  • the capillary junction 904 at the distal end of the metering capillary 902 is overcome, and sample from metering capillary 902 fills incubation chamber 910 ( Figures 27F and 27G). A portion of the sample wicks into U- shaped capillary 914 to the level of the sample in incubation chamber 910 ( Figure 27G).
  • the sample is incubated for a time sufficient for maximum saturation binding of the component in the sample that specifically bind to the specific binding species.
  • a third rotational speed f 3 that is greater than the second rotational speed f 2 , typically in the range of 100-1500rpm, the capillary junction 908 is overcome, and wash buffer from reservoir 916 flows through capillary 909, capillary 906, and into incubation chamber 910. Wash buffer fluid flow forces the sample through U-shaped capillary 914 and into waste reservoir 915 ( Figures 27 H through 27J).
  • sacrificial valves 922 are released to permit wash buffer fluid flow.
  • a fourth rotational speed f 4 that is greater than the third rotational speed f 3 , typically in the range of 100-2000rpm
  • the capillary junction 919 is overcome, and reagent buffer from reservoir 917 flows through capillary 918, capillary 920, capillary junction 908, capillary 906, and into incubation chamber 910.
  • Reagent buffer fluid flow forces the wash buffer through U-shaped capillary 914 and into waste reservoir 515 ( Figures 27 H through 27J).
  • sacrificial valves 922 are released to permit reagent buffer fluid flow.
  • the reagent buffer contains a chromogen or other developing agent for detection of specific binding in incubation chamber 910.
  • Temperature control elements are provided to control the temperature of the platform.
  • the invention provides heating elements, specifically resistive heating elements, and elements for detecting temperature at specific positions on the platform. Heating devices are preferably arrayed to control the temperature of the platform over a particular and defined area, and are provided having a steep temperature gradient with distance on the platform from the heater.
  • Certain resistors including commercially- available resistive inks (available from Dupont) exhibit a positive temperature coefficient (PTC), i.e., an increase in resistance with increasing temperature.
  • PTC positive temperature coefficient
  • connection to a power source is made by first printing parallel silver conductors followed by printing the PTC ink between the conductors as shown in Figure 29.
  • a resistive heating element comprises a conductive ink connected with electrical contacts for activation of the heater, and resistive inks applied between the conductive ink and in electrical contact therewith, wherein application of a voltage (direct or alternating cu ⁇ ent) between the conductive inks results in current flow through the resistive inks and production of heat.
  • resistive inks used in the resistive heating elements of this invention The first is a standard polymer thick film ink, such as Dupont 7082 or Dupont 7102 ink. These inks produce a surface temperature that is not self-limiting, and the temperature resulting from the use of these inks is dependent primarily on the magnitude of the applied voltage.
  • PTC inks show increase resistivity with increasing voltage, so that surface temperature is self-limiting because the amount of heat-producing current goes down as the applied voltage goes up.
  • PTC inks are characterized as having a particular temperature where this self-limiting property is first exhibited; at voltages that produce temperatures less than the critical temperature, the amount of heat is dependent on the magnitude of the applied voltage.
  • Resistive inks useful according to the invention include Dupont 7082, 7102,
  • Conductive inks useful according to the invention include Dupont 5028, 5025, Acheson 423SS, 426SS and SS24890, and other equivalent commercially available conductive inks.
  • Dielectric layers advantageously comprise dielectric inks such as Dupont
  • Insulation can also be achieved using pressure sensitive transfer adhesive such as 7952MP (3M Co.), or a pressure sensitive transfer adhesive deposited onto a polyester carrier layer such as 7953MP (3M Co.) or thermoplastic bonding films such as 3M 406, 560 or 615.
  • Resistive heaters of the invention are advantageously used to incubate fluids at a stable temperature and for melting sacrificial valves as described below, and also for thermal cyclic.
  • Resistive and conductive inks are preferably screen-printed using methods and techniques well known in the art. See Gilleo, 1995, Polymer Thick Film (Van Nostrand Reinhold). Inks are typically screen printed to a thickness of about 10 microns; however, repetitive screen printing of resistive inks can be used to deposit thicker layers having reduced resistances. Both conductive and resistive inks are heat cured, typically at between 110 °C and 120 °C for about 10 minutes. The outline of this printing process is shown in Figure 30. Importantly, each of the layers must be correctly registered with one another for resistive heating to be provided. Heaters can be screen printed to any required size; a minimum area for a screen-printed heater has been determined to be about 0.25mm 2 (0.5mm x 0.5mm).
  • the ability to tailor the resistance (and hence the temperature profile) of the resistive heaters using choice of ink formulation and reprinting of heater circuits provides control of the final electrical and thermal properties of the resistive heating elements of the invention.
  • the resistance can also be controlled through connection of series and parallel configurations of resistive elements.
  • the particular circuits shown in Figure 31 allow for many parallel resistive elements per unit area; other configurations can be chosen for other applications.
  • sacrificial valve is intended to encompass materials comprising waxes, plastics, and other material that can form a solid or semi-solid fluid-tight obstruction in a microchannel, capillary, chamber, reservoir or other microfluidics component of the platforms of the invention, and that can be melted or deformed to remove the obstruction with the application of heat.
  • Sacrificial valves are preferably made of a fungible material that can be removed from the fluid flow path.
  • said sacrificial valves are wax valves and are removed from the fluid flow path by heating, using any of a variety of heating means including infrared illumination and most preferably by activation of resistive heating elements on or embedded in the platform surface as described herein.
  • heating any of a variety of heating means including infrared illumination and most preferably by activation of resistive heating elements on or embedded in the platform surface as described herein.
  • the term "wax" is intended to encompass any solid, semi-solid or viscous liquid hydrocarbon, or a plastic. Examples include mondisperse hydrocarbons such as eicosane, tetracosane and octasone, and polydisperse hydrocarbons such as paraffin.
  • the wax recrystallization chamber is fluidly connected with the microchannel, capillary or other fluidic component of the microsystems platforms of the invention that was occluded by the wax sacrificial valve.
  • the wax recrystallization chamber is a widening of the microchannel, capillary or other fluidic component of the microsystems platforms of the invention so that recrystallized wax can harden on the walls of the microchannel, capillary or other fluidic component of the microsystems platforms of the invention with enough distance between said walls that the recrystallized wax does not re-occlude the microchannel, capillary or other fluidic component of the microsystems platforms of the invention.
  • the heating element most preferably the resistive heating element of the invention, extends past the site of the wax valve and overlaps at least a portion of the wax recrystallization chamber, thereby retarding the propensity of the wax valve to recrystallize.
  • wax valves to recrystallize can be exploited to create a wax valve at a particular location in a microchannel, capillary or other fluidic component of the microsystems platforms of the invention.
  • a particular location can be kept below a threshold temperature by failing to apply heat at that location, and a wax valve material can be mobilized from a storage area on a platform by heating and them allowed to flow under centripetal acceleration to a particularly "cold" site where a wax valve is desired.
  • wax valves in this regard is that the proper positioning an activation of resistive heater elements enables flexibility in choosing when and whether a particular microchannel, capillary or other fluidic component of the microsystems platforms of the invention is to be occluded by a wax sacrificial valve.
  • the sacrificial valves of the invention comprise a cross-linked polymer that displays thermal recover, most preferably a cross- linked, prestressed, semicrystalline polymer; an example of a commercially available embodiment of such a polymer is heat recoverable tubing (#FP301H, 3M Co.,
  • the sacrificial valves of the invention comprise a thin polymeric layer or barrier dividing two liquid-containing microchannel, capillary or other fluidic component of the microsystems platforms of the invention, that can burst when sufficient temperature and/ or pressure is applied.
  • a screen-printed resistive heater element is itself a valve.
  • the resistive heater element is screen-printed on a substrate such as polyester that divides two liquid-containing microchannel, capillary or other fluidic component of the microsystems platforms of the invention.
  • a substrate such as polyester that divides two liquid-containing microchannel, capillary or other fluidic component of the microsystems platforms of the invention.
  • localized application of heat using a resistive heating element is used to melt the substrate dividing the liquid- containing microchannel, capillary or other fluidic component of the microsystems platforms of the invention.
  • the two liquid-containing microchannel, capillary or other fluidic component of the microsystems platforms of the invention are positioned in adjacent layers through the vertical thickness of the platform.
  • the screen-printed resistive heater elements of this invention provide localized application of heat to a microsystems platform.
  • the degree of localization achieved using these resistive heating elements is sufficient to provide for the placement of two adjacent sacrificial valves separated by a distance of 0.15cm.
  • the invention also provides a specialized rotor spindle for making electrical connection through a spindle to a rotating structure.
  • An example of the specialized spindle structure of the invention is shown in Figure 1.
  • the spindle is provided with a series of electrically conductive rings, each of which are electrically isolated from the others. Each conductive ring is led to contacts that impinge upon circuits resident on the rotating structure. Electrical power or data signals are delivered to and from the apparatus by means of conductive brushes that contact the conductive rings. Signals may be conducted while the apparatus is rotating or when stationary.
  • Rotating structures such as centrifugal rotors and the microsystems platforms of this invention containing internal channels and reservoirs are used to control fluid movement.
  • centrifugal analyzers Such disks and rotors are known in the art as centrifugal analyzers and have been used to separation chemical and biochemical species as well as enabling the synthesis of chemical and biochemicals.
  • a limitation of conventional centrifugal analyzers is that those analytical or synthetic procedures that required electrical input on the rotating structure itself were not enabled.
  • mechanical and non-mechanical processes were used to regulate fluid movement in centrifuge rotors and other centripetally-motivated devices. Mechanical processes included valve actuation and pumping liquid toward the center of rotation; non-mechanical processes included heating, cooling, electrophoresis, and sensing through combinations of optical, electrical, chemical and biological means.
  • the mechanical and non-mechanical processes known in the prior art were not capable of precisely regulating fluid movement when compared with equivalent, non-rotating devices that used electrical power.
  • Rotating structures permit precise control of fluid movement.
  • Conventional centrifugal fluid control is restricted to outward flow in a radial direction that is primarily regulated through liquid placement (such as volume, positional radius, and liquid height), channel geometry (including considerations of channel radius) and rotational rate.
  • Temperature control is achieved over the entire rotor, by refrigerating the centrifugal compartment, for example.
  • many chemical and biological processes require precisely controlled elevated temperature for optimal performance; for example, enzymatic reactions such as those used in immunoassays often have temperature optima of about 37 °C.
  • In vitro amplification reactions (such as polymerase chain reaction) require even more elevated temperatures, such as cycling between 70 °C and 95°C.
  • a rotating structure presents unique difficulties in terms of thermal control, particularly heating.
  • Conventional options included placement of the centrifuge rotor inside of an oven, subjecting one area to infrared radiation, and clamping the device between thermally controlled platens.
  • Each of these approaches present difficulties.
  • Ovens have unpredictable thermal gradients that prevent tight control of temperature 'in the interior of a rotating structure.
  • Infrared presents similar difficulties, while clamping precludes spinning while heating unless the entire heating apparatus is also spun. All of these approaches suffer from an inability to directly measure the temperature inside the rotating structure.
  • This invention provides a solution to this problem, specifically by enabling the transmission of electrical signals between a rotating structure such as a centrifuge rotor or microsystem platform of this invention and a fixed position electrical source.
  • Electrical devices that can be advantageously controlled on a spinning rotor include temperature regulation devices, sensors, electrophoretic devices, integrated circuits and mechanical valves. These structures in turn permits a wider array of chemical processes to be performed on a disk than those permitted with current technology.
  • the invention provides a specialized rotor spindle for transmitting electrical signals between a rotating centrifuge rotor and a stationary electrical source as follows; the structure of the spindle will be best appreciated with reference to Figure 1.
  • a plurality of electrically conductive posts Into an electrically non-conductive plate is embedded a plurality of electrically conductive posts.
  • the non-conductive plate is made of any insulating, electrically non-conductive material, including most preferably hardened plastics, rubber, WHAT ELSE??, and other electrical insulators and non-conductors.
  • the electrically conductive posts are made from any electrically conducting metal, including copper, aluminum, WHAT ELSE???, and are preferably spring-loaded from the bottom of the non-conductive plate.
  • a mechanical spindle is positioned on the side of the non-conductive plate opposite to the side of the plate containing the conductive posts.
  • This plate is referred to as a spindle plate.
  • a number of conductive disks equal to the number of conductive posts are layered on the spindle plate.
  • the electrical contacts in the conductive plate are arranged so that each conducive plate is in electrical contact with a single conductive post.
  • the term "in electrical contact” is intended to mean that an electric current can be produced through the electrical contacts at voltages (direct or alternative current) that may be effectively achieved in a centrifugal apparatus as described.
  • Each conductive disk is similarly electrically isolated from other disks with interspersed sheets or disks of nonconducting material, preferably the same non- conductive material used to make the non-conductive plate.
  • the last layer positioned farthest way from the spindle plate is a nonconductive layer and is arranged so that the tips of the conductive posts are at the surface or emerge from this plate.
  • This final plate containing the ends of the conductive posts is referred to as the contact plate.
  • An assembled unit, consisting of baseplate, posts, conductive element within the stack, is referred to as a signal channel. Typically one channel will be reserved as a common or ground channel.
  • the contact stack is held within a chassis that permits a series of electrically conductive brushes to impinge upon each of the conductive disks of the contact stack.
  • These brushes are arranged such that an electric signal can be propagated from one brush to one contact on the contact plate independently of electrical signals conducted from any other brush.
  • the spindle is held in a spring-loaded bearing assembly such that the contact stack can rotate freely and present resistive pressure when pressed from below.
  • the assembled unit is referred to as an electronic spindle.
  • a centrifugal rotor or microsystems platform disk of this invention containing electrical contacts is aligned with the contacts on the contact plate of the electronic spindle in such a fashion that the electrical contact on the rotor are in electrical contact with the conductive posts on the contact plate of the electronic spindle.
  • An electric signal can be propagated from the brushes of the electronic spindle to the contacts on the disk using the electronic spindle, most importantly when the rotor is spinning. Contact between the contact plate and the rotor is maintained by positive pressure provided by the spring-loaded bearing assembly and conductive posts. Electrical devices resident on the disk can then be monitored or controlled by signals through the electronic rotor.
  • the rotor and the spindle contain complementary mechanical components that precisely align the rotor on the spindle to ensure proper electrical contact between the contacts on the rotor and the conductive posts on the spindle.
  • thermal control structures resident on a rotating disk may be controlled through the electronic spindle.
  • a resistive heating element prepared in a rotor as described herein is prepared having electrical contact leads positioned to be brought into electrical contact with one signal channel and the common channel of the electronic spindle.
  • a thermistor element as described herein is placed on the disk in close proximity to the resistive heating element to enable thermistor response to be proportional to the temperature of the resistive heating element.
  • the temperature of the heating element will be a function of the voltage and current applied to the disk through the electronic rotor and the speed at which the disk is being rotated (which will promote convective cooling).
  • a thermal profile may be accurately monitored through the thermistor response and temperature controlled through adjustment of heating voltage and disk speed using the electronic spindle of the invention.
  • the polymerase chain reaction (“PCR"') can be carried out on a rotating disk through thermal control using the electronic spindle.
  • a reaction chamber on a rotating disk is provided with the reagents necessary for PCR to provide a reaction mixture (see Saiki et al. , 1985 , Science : - ).
  • This reaction chamber is in close proximity to a heater and thermistor as described herein to enable thermistor output to be proportional to the temperature of the reaction mixture.
  • the reaction chamber is subjected to a cycling temperature profile sufficient to permit amplification of the template. Since the rate and precision at which a temperature change can be accomplished is a determining factor in the success of PCR amplification, control of voltage and speed of rotation as enabled using the electronic spindle of the invention permits PCR to be performed on a rotating platform.
  • enzyme-requiring assays such as enzyme linked immunosorbant assay (“ELISA")
  • ELISA assays consist of antibody/antigen binding interactions followed by the conversion of a colorimetric or radioactive substrate into a detectable product. Detection is carried out through electrochemical or optical processes, depending the nature of the detectable product. Both antibody/antigen binding and enzymatic reactions have optimal temperatures (typically 37 °C) that can be achieved on a rotating platform using the resistive heater/ thermistor pair described above using the electronic spindle.
  • Electromechanical valves known in the prior art can also be incorporated in a rotating platform of centrifuge rotor using the electronic spindle of the invention.
  • Mechanical, electrolytic or thermal valves on a rotating disk have been described (for example, in co-owned and co-pending U.S. Serial No. 08/761,063).
  • the fractionation of complex mixtures can be carried out on a rotating platform.
  • complex biological mixtures such as milk or blood are subjected to centrifugal force to effect separation of constituent parts.
  • Electromechanical, electrolytic or thermal valves can be opened, for example, to withdraw a fractionated supernatant away from a sedimented layer and into an adjacent chamber.
  • Sample fractionation can be accomplished using repetitive sedimentation at different rotational rates, with appropriate valve activation used to partition portions of the fractionated to separate reservoirs or other compartments on the rotor. Upon fractionation the constituents may then be subjected to other processes such as PCR, immunoassay or electrophoresis.
  • Another application of the electronic spindle of the invention is to activate sensors on the rotating platform.
  • sensors for detecting pH are controlled and monitored through the electronic spindle.
  • FIG. 1 A microsystems platform provided by the invention and specifically designed for performing antibiotic assays is illustrated in Figures 1 and 2.
  • Disk embodiments of the platforms of the invention were fashioned from machined acrylic and injection-molded polycarbonate.
  • the overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk was mounted on the spindle of a rotary device.
  • the thickness of the disc ranged from about 0.9mm to about 1.5mm.
  • the components of the antibiotic array were prepared as follows.
  • An entry port 201 having a depth in the platform surface from about 0.75mm and lateral dimensions of from about 0.2cm to about 2cm were constructed on the platform, and designed to accommodate a volume of about 60 ⁇ L.
  • This entry port was fluidly connected with an array of eight metering capillaries 202 having a square cross-sectional diameter of about 0.5mm and proximal ends rounded with respect to entry port 201; the length of this metering capillary array was sufficient to contain a total volume of about 20 ⁇ L.
  • the entry port was also constructed to be fluidly connected with an overflow capillary 203 having a cross-sectional diameter of from about 0.02mm to about 0.75mm and proximal ends rounded with respect to entry port 201.
  • the overflow capillary was fluidly connected with an overflow chamber 205 having a depth in the platform surface of about 0.75mm, greater than the depth of the overflow capillary 203.
  • Metering capillary 202 was fluidly connected to fluid chamber 204 having a depth in the platform surface of about 0.63mm and greater than the depth of the metering capillary 202.
  • Each of the overflow and fluid chambers was also connected with air ports or air channels, such as 211, that have dimensions of about 0.25mm deep and permitted venting of air displaced by fluid movement on the platform.
  • a capillary junction 212 that is about 0.75mm deep is present in the air channel to prevent fluid flow into the air channel.
  • Entry port 201 was positioned on the platform from about 2cm from the center of rotation.
  • Metering capillary 202 extended about 1 cm from entry port 201.
  • the extent of the length of overflow capillary 203 was at least about 20%o greater than the extent of the length of metering capillary 202.
  • the position of fluid chamber 204 was from about 3.2cm from the center of rotation, and the position of overflow chamber 205 was thus from about 5 cm from the axis of rotation.
  • the fluid chamber 204 acted as a capillary barrier that prevents fluid flow from metering capillary 202 at a first, non-zero rotational speed f, sufficient to permit fluid flow comprising overflow from the entry port 201 through overflow capillary 203 and into overflow chamber 205.
  • This capillary boundary was constructed to be overcome at a second rotational speed f 2 (where f 2 > fi).
  • Fluid chamber 204 was fluidly connected to capillary 206 that was 0.25mm deep and had a cross-sectional diameter of about 0.5mm and was connected to holding chamber 207. Holding chamber 207 had a depth in the platform surface of 0.75mm, greater than the depth of capillary 206.
  • Fluid chamber 204 is accompanied by fluid flow through capillary 206 into holding chamber 207.
  • Holding chamber 207 was fluidly connected by way of capillary 208, having a square cross-sectional diameter of about 0.25mm and was connected with read chamber 210, having a depth in the platform surface of about 0.75mm, greater than the depth of capillary 208.
  • a sacrificial valve 213 was placed as shown in the channel 209. As illustrated in Figures 3 A through 3J, in the use of this platform an imprecise volume (ranging from 20-60 ⁇ L of fluid) of a fluid was applied to the entry port 201 ( Figure 3 A).
  • the fluid wicked into air channel 211 and was stopped by capillary junction 212. Fluid also wicked into metering capillary 202 and overflow capillary 203. Fluid flowed through the metering capillary 202 and overflow capillary 203 at no rotational speed until the fluid reached capillary junctions at the junction between metering capillary 202 and fluid chamber 204 and overflow capillary 203 and overflow chamber 205 ( Figures 3B and 3C).
  • Metering capillary 202 was constructed to define a precise volume from about 20- 60 ⁇ L of fluid between entry port 201 and the capillary junction at fluid chamber 204, which was designed to be at least the amount of the fluid placed by the user in entry port 201.
  • the platform was spun at a first rotational speed f ]5 ranging from of 175rpm, which was sufficient for this microfluidics array having an entry port 201 with a depth of 0.6mm, metering capillary 202 with dimensions of 0.5mm x 0.5mm in cross-section and a length of 2.2-3.8cm from the center of rotation and an overflow capillary 203 with dimensions of 0.5mm x 0.5mm in cross-section and a length of 5.4cm from the center of.
  • This embodiment of the microfluidics platforms of the invention was designed to use the carboxypeptidase inhibition assay described above for detecting beta-lactam antibiotics.
  • the extent of chromogen production was detected in the read chamber, and related to the presence of antibiotics in the sample by comparison with samples tested in the absence of antibiotic.
  • the amount of antibiotic in a test sample was determined using this platform of the invention.
  • a two-step assay disk of the invention was provided as shown in Figures 4 and 5.
  • Disk embodiments of the platforms of the invention were fashioned from machined acrylic.
  • the overall disc dimensions included an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk was mounted on the spindle of a rotary device.
  • the thickness of the disc ranged from about
  • the working fluid volume for reaction with reagents was about 20 ⁇ L.
  • This two-step assay array were prepared as follows.
  • An entry port 301 having a depth in the platform surface of about 0.75mm and lateral dimensions of from about 0.2cm to about 2cm was constructed on the platform, and designed to accommodate a volume of about 60 ⁇ L.
  • This entry port was fluidly connected with a multiplicity of entry capillaries 302 having a square cross-sectional diameter of about 0.5mm, having a depth of 0.5mm and proximal ends rounded with respect to entry port 301; the length of this entry capillary array was sufficient to contain a total volume of about 20 ⁇ L.
  • the entry capillaries 302 were fluidly connected to fluid chamber 303 having a depth in the platform surface of about 0.6mm, wherein the depth was greater than the depth of the entry capillary 302.
  • Each of the fluid chambers of this aspect of the invention was also connected with air ports or air channels, such as 311, that have dimensions of about 0.25mm deep and permitted venting of air displaced by fluid movement on the platform.
  • a capillary junction 312 that was about 0.75mm deep was present in the air channel to prevent fluid flow into the air channel.
  • the fluid chamber 303 was also constructed to be fluidly connected with an overflow capillary 304 having a cross-sectional diameter of from about 0.02mm to about
  • the overflow capillary was fluidly connected with an overflow chamber 306 having a depth in the platform surface of about 0.75mm, greater than the depth of the overflow capillary 304.
  • Entry port 301 was positioned on the platform about 1cm from the center of rotation. Entry capillaries 302 extends about 2cm from entry port 301. The position of a first fluid chamber 303 was from about 3cm from the center of rotation. The first fluid chamber 303 acted as a capillary barrier that prevented fluid flow from entry capillary 302 at zero rotational speed. Movement of fluid from entry port 301 through entry capillaries 302 and into the first fluid chamber 303 was achieved by rotation at a first, non-zero rotational speed f,. Displacement of fluid into the first fluid chamber 303 was accompanied by fluid filling of channel 305 that was fluidly connected with the first fluid chamber 303 and was positioned at the most radially distal point of the first fluid chamber.
  • Channel 305 was fluidly connected with a second fluid chamber 307 and formed a capillary boundary between channel 305 and chamber 307. This capillary boundary was constructed to be overcome at second rotational speed f 2 (where f 2 > f j ).
  • First fluid chamber 303 was also fluidly connected to overflow capillary 304 that was 0.25mm deep and had a cross-sectional diameter of about 0.5mm and that extended from about 1cm to about 5cm and was connected to overflow chamber 306.
  • Overflow chamber 306 had a depth in the platform surface equal to that of overflow capillary 304, so that there was no capillary boundary between overflow capillary 304 and overflow chamber 306.
  • Overflow capillary 304 was positioned in the first fluid chamber 303 at a point radially less distant from entry port 301 than channel 305, thereby defining a volume in the fluid chamber between the position of the overflow capillary 304 and the most radially distant extent of the said first fluid chamber.
  • Second fluid chamber 307 was further fluidly connected through channel 308 to a small pocket or capillary junction 309.
  • Channel 308 having a cross-sectional diameter of about 0.25mm and that extended from about 0.2cm to about 20cm, was fluidly connected to a third fluid chamber 310, having a depth in the platform surface of about 0.75mm, that was greater than the depth of capillary 308.
  • Air recirculation channels 311 that had dimensions of about 0.25mm deep provided pathways for air displaced by fluid movement, while capillary junctions 312 that were about 0.75mm deep prevent fluid from entering the air channels.
  • a sacrificial valve 313 was placed as shown in Figure 5 in the channel 309.
  • a valve 314 was placed in channel 305 to control fluid movement from the first fluid chamber 303 to the second fluid chamber 307.
  • a sacrificial valve 314 was placed at the junction of channel 305 and the second fluid chamber 308, which sacrificial valve was released to permit fluid flow through channel 305 and into the second fluid chamber 308.
  • fluid flow can be achieved at either f, or f 2 rotational velocity.
  • a microsystems platform provided by the invention and specifically designed for separating vertebrate blood cells and components is illustrated in Figures 7 through 9.
  • FIG. 8 It will be understood by a comparison of Figures 7 and 8 that the center of the platform 11 is at the top of Figure 8, and the edge or lateral extent of the platform is at the bottom of Figure 8, illustrated by a curved line.
  • Rotation of the blood separation array on platform disks of the invention can be in either direction, although rotation in a consistent, particular direction is preferred.
  • Disk embodiments of the platforms of the invention were fashioned from machined acrylic.
  • the overall disc dimensions included an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device.
  • the thickness of the disc ranged from about 0.9mm to about 1.5mm.
  • the working fluid volume for reaction with reagents was about
  • An entry port 401 having a depth in the platform surface of about 0.5mm and lateral dimensions of about 0.5cm was constructed on the platform, and designed to accommodate a volume of about 20 ⁇ L.
  • This entry port was fluidly connected to an entry capillary 402, having a cross-sectional diameter of about 0.5mm and having a depth of 0.5mm; the length of this entry capillary was sufficient to contain a total volume of about 20 ⁇ L.
  • Entry capillary 402 was further fluidly connected to a separation column 403 having a cross- sectional diameter of about 1.25mm, a depth of about 0.75mm, and a length of this separation column was sufficient to contain a total volume of about 15 ⁇ L.
  • This separation column was also fluidly connected with a passage 411 to overflow chamber 404.
  • Passage 411 had a cross-sectional diameter of about lmm, a depth of from about 0.5mm, and a length of 2mm.
  • Overflow chamber 404 has a depth of about 0.5mm.
  • a small capillary exit 406 was also fluidly connected with separation chamber 403, having a cross-sectional diameter of about 0.125mm, a depth of about 0.125mm, and a length of about 0.75mm. This capillary was arranged to traverse a direction radially more proximal to the axis of rotation than the insertion point with separation column 403. This small capillary 406 terminated in a capillary junction 407 that was fluidly connected with capillary 408, extending in a radial direction to decant chamber
  • a sacrificial valve 413 is positioned in capillary 406 at the juncture with capillary junction 407.
  • Capillary 408 had a cross-sectional diameter of about 0.25mm, a depth of about 0.25mm, and a length of about 3.5mm. This capillary was arranged in a radially outward direction between capillary junction 407 and decant chamber 405.
  • Passage 411 was positioned on separation column 403 to be significantly more proximal to the axis of rotation than the insertion point of small capillary 406.
  • Air displacement channels 409 that have dimensions of about 0.25mm deep permitted venting of air displaced by fluid movement on the platform.
  • Capillary junctions 410 that were about 0.75mm deep were present in the air channels to prevent fluid flow into the air channels.
  • a blood separation disk of the invention is provided as shown in Figures 10 and 11. It will be understood that, as in Example 1, in Figure 10, the arrangement of one separation array 15 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
  • Disk embodiments of the platforms of the invention are fashioned from machined acrylic.
  • the overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device.
  • the thickness of the disc ranged from about 0.9mm to about 1.5mm.
  • the working fluid volume for reaction with reagents is about 25 ⁇ L.
  • An entry port 501 having a depth in the platform surface from about 0.75mm and lateral dimensions of from about 0.5cm is constructed on the platform, and designed to accommodate a volume of about 20 ⁇ L.
  • This entry port is fluidly connected with a first array of metering capillaries 502 and a second array of metering capillaries 503, wherein each of the capillaries has a cross-sectional diameter of about 0.5mm.
  • the length of the second metering capillary array 503 is longer than that of the first metering capillary array 502.
  • the first metering capillary array 502 is fluidly connected with a ballast chamber 507, having a depth in the platform surface of about 0.75mm and greater than the depth of the first metering capillary array 502, wherein the first metering capillary array 502 forms a capillary junction between the array and the ballast chamber.
  • the second capillary a ⁇ ay 503 is fluidly connected with capillary junction 506.
  • the entry port is also constructed to be fluidly connected with an overflow capillary 504 having a cross-sectional diameter of about 0.5mm and proximal ends rounded with respect to entry port 501.
  • the overflow capillary is fluidly connected with an overflow chamber 505 having a depth in the platform surface of about 0.75mm, greater than the depth of the overflow capillary 504.
  • Each of the overflow and fluid chambers is also connected with air ports or air channels, such as 514, that have dimensions of about 0.25mm deep and permit venting of air displaced by fluid movement on the platform. Capillary junctions 516 that is about 0.75mm deep are present in the air channels to prevent fluid flow into the air channels.
  • Entry port 501 is positioned on the platform from 1cm to 20cm from the center of rotation.
  • Metering capillary array 502 extends from entry port 501 from about 0.6cm.
  • Metering capillary array 503 extends from entry port 501 from about 1.9cm.
  • the extent of the length of metering capillary array 503 is about 20%> longer than metering capillary array 502, and the extent of the length of overflow capillary 504 is at least about 20% greater than the extent of the length of either the first metering capillary array 502 or the second metering capillary array 503.
  • ballast chamber 507 is about 2.8cm from the center of rotation
  • the position of capillary junction 506 is about 3.8cm from the center of rotation
  • the position of overflow chamber 505 is thus about 5cm from the axis of rotation.
  • the ballast chamber 507 acts as a capillary barrier that prevents fluid flow from the first metering capillary array 502 at a first, non-zero rotational speed fj sufficient to permit fluid flow comprising excess blood overflow from the entry port 501 through overflow capillary 504 and into overflow chamber 505.
  • Capillary junction 506 is a capillary barrier that prevents fluid flow from the second metering capillary array 503 at said first, non-zero rotational speed f, sufficient to permit fluid flow comprising excess blood overflow from the entry port 501 through overflow capillary 504 and into overflow chamber 505.
  • These capillary boundaries are constructed to be overcome at a second rotational speed f 2 (where f 2 > fj).
  • Ballast chamber 507 is fluidly connected to capillary 510 that is 0.25mm deep and has a cross-sectional diameter of about 0.65mm and that extends about 5cm and is connected to capillary junction 511.
  • a sacrificial valve 518 is positioned at the exit of ballast chamber 507 at the juncture with capillary junction 511.
  • capillary 510 is fluidly connected with a sacrificial valve 515.
  • fluid flow is achieved at rotational speed f 2 with removal of the sacrificial valve.
  • capillary junction 511 are further fluidly connected with channel 512 which is about 0.25mm deep and has a cross-sectional diameter of about 0.25mm and that extends about 3cm.
  • Channel 512 is fluidly connected with separation chamber 509 at a point most distal from the axis of rotation.
  • Second metering capillary array 503 is fluidly connected with capillary junction
  • Capillary junction 506 is further fluidly connected to channel 508, which is further fluidly connected to separation chamber 509.
  • Channel 508 is about 0.25mm deep and has a cross-sectional diameter of about 0.25mm.
  • Separation chamber 509 is about 0.75mm deep and has a cross-sectional diameter of about 5mm, and is positioned from about 40cm from the center of rotation.
  • Separation chamber 509 is fluidly connected with decant channel 517 at a point close to the chamber's most axis-proximal extent.
  • Decant channel 517 is about 0.25mm deep and has a cross-sectional diameter of about 0.25mm and extends about 4.3cm to about 5cm.
  • Decant channel 517 is fluidly connected with decant chamber 514, which is about 0.75mm deep and has a cross-sectional diameter of about 5mm, and is positioned from about 50cm to about 80cm from the center of rotation..
  • an imprecise volume (about 25 ⁇ L of fluid) of blood is applied to the entry port 501 ( Figure 12 A).
  • Blood enters the each of the metering capillary arrays 502 and 503 and stops at the capillary junction between metering capillary array 502 and ballast chamber 507 and between metering capillary 503 and capillary junction 506 ( Figures 12B and 12C). Blood also enters and fills overflow capillary 504, stopping at the capillary junction with overflow chamber 505.
  • the capillary junction between the first metering capillary array 502 and ballast chamber 508 is overcome, and blood from the first metering capillary array fills ballast chamber 508 ( Figure 12F).
  • capillary junction 506 is overcome, and blood from second metering capillary array 503 enters separation chamber 509 ( Figure 12F).
  • the volume of blood in second metering capillary a ⁇ ay 503 is insufficient to fill separation chamber 509 to the level of insertion of decant channel 517.
  • EXAMPLE 5 Mixing Arrays
  • a microsystems platform provided by the invention and specifically designed for performing mixing of equal volumes of different liquid samples is illustrated in Figure 13.
  • the arrangement of one assay a ⁇ ay 15 on a disk 11 is shown; a multiplicity of such a ⁇ ays can be advantageously a ⁇ anged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
  • the components of the mixing array are shown in greater detail in Figure 14.
  • Disk embodiments of the platforms of the invention were fashioned from machined acrylic.
  • the overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device.
  • the thickness of the disc ranged from about 0.9mm to about 1.5mm.
  • the working fluid volume was about 50 ⁇ L.
  • the components of the mixing array were prepared as follows. Entry ports 601 having a depth in the platform surface of about 0.5mm and lateral dimensions of about lcm to about 5cm were constructed on the platform, and designed to accommodate a volume of about 5-50 ⁇ L.
  • Each entry port was fluidly connected with one of a paired a ⁇ ay of metering capillaries 602 having a square cross-sectional diameter of about 0.5mm and proximal ends rounded with respect to entry port 601; the length of each metering capillary a ⁇ ay was sufficient to contain a total volume of about 25 ⁇ L.
  • Metering capillaries 602 were fluidly connected to a curved capillary ba ⁇ ier 603 having a depth in the platform surface of about 0.25mm that was greater than the depth of metering capillaries 602.
  • the capillary ba ⁇ ier 603 and other fluid components of the mixing a ⁇ ay were also connected with air channels 608, that have dimensions of about
  • capillary junctions 609 that were about 0.75mm deep were present in the air channels to prevent fluid backflow into the air channel.
  • Capillary ba ⁇ ier 603 was fluidly connected by a na ⁇ ow capillary channel 604 to mixing chamber 605, which was fluidly connected with channel 610, which was further connected with mixed fluid receiving chamber 606.
  • capillary 604 comprises a sacrificial valve 612.
  • Capillary channel 604 was about 0.25mm deep and had a cross-sectional diameter of about 0.25mm and that extends about 0.2cm.
  • Mixing chamber 605 was about 0.75mm deep and had a cross-sectional diameter of about 2mm, and was positioned about 4cm from the center of rotation.
  • Capillary channel 610 was about 0.25mm deep and had a cross-sectional diameter of about 0.25mm and that extended from about 0.2cm to about 30cm.
  • Mixing chamber 605 was constructed such that the insertion point of capillary channel 604 and the insertion point of capillary channel 610 were offset at opposite ends of the mixing chamber. As a consequence, fluid flowing through capillary channel 604 was forced to encounter the opposite wall of mixing chamber 605 before fluid flow can proceed through capillary channel 610. This results in the creation of turbulence in the mixed laminar fluid stream in capillary channel 604 caused by the conjoint flow of fluid from the first and second metering channels without appreciable mixing. The turbulence created by the structure of mixing chamber 605 was sufficient to disrupt laminar flow and cause fluid mixing in the chamber prior to continued fluid flow through capillary channel 610 and into mixed fluid receiving chamber 606.
  • Mixed fluid receiving chamber 606 was about 0.75mm deep and had a cross-sectional diameter of about 5mm, and was positioned from about lcm to about 30cm from the center of rotation.
  • FIG. 17 An additional embodiment of the microsystems platform provided by the invention and specifically designed for performing mixing of equal volumes ( Figures 17 through 18) or unequal volumes (Figures 19 through 21) of different liquid samples.
  • the a ⁇ angement of one assay a ⁇ ay 17 on a disk 11 is shown; a multiplicity of such a ⁇ ays can be advantageously a ⁇ anged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
  • the components of the mixing a ⁇ ay are shown in greater detail in Figures 17 and 20.
  • Disk embodiments of the platforms of the invention are fashioned from machined acrylic.
  • the overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device.
  • the thickness of the disc ranged from about 0.9mm to about 1.5mm.
  • the working fluid volume is about 40 ⁇ L in each fluid reservoir.
  • Fluid reservoirs 651 and 652 (701 and 702) are constructed on the platform, having a depth in the platform surface from about 0.75mm and lateral dimensions of lcm, and wherein fluid reservoirs 651 and 652 are designed to accommodate equal volumes of fluid (about 50 ⁇ L); (for unequal volumesn fluid reservoir 701 is designed to accommodate a volume of about 45 ⁇ L, and fluid reservoir 702 is designed to accommodate a volume of about 5 ⁇ L, wherein the volume of fluid reservoir 702 is less than the volume of fluid reservoir 701.)
  • the viscosity of the fluid in the fluid reservoirs may differ, so that mixing produces a mixed fluid of intermediate viscosity.
  • Each fluid reservoir is fluidly connected with a capillary channel 653 and 654 (703 or 704) to capillary junction 655 (705).
  • Each capillary channel is about 0.5mm deep and having a cross-sectional diameter of about 0.5mm and that extends about 5cm.
  • Capillary junction junction 655 (705) has a depth in the platform surface of about 0.75mm that is greater than the depth of capillaries 652 and 653 (703 and 704).
  • capillaries 652 and 653 (703 and 704) comprise a sacrificial valve 662 (712). Use of said sacrificial valves can be used in addition to or in place of capillary junction 655 (705).
  • the fluid components of the mixing a ⁇ ay are also connected with air channels 660 (710), that have dimensions of about 0.25mm deep and permit venting of air displaced by fluid movement on the platform.
  • air channels 660 (710) that have dimensions of about 0.25mm deep and permit venting of air displaced by fluid movement on the platform.
  • capillary junctions 661 (711) that are about 0.75mm deep are present in the air channels to prevent fluid backflow into the air channel.
  • Capillary junction 655 (705) is fluidly connected by a na ⁇ ow capillary channel 656 (706) to mixing chamber 657 (707), which is fluidly connected with channel 658 (708) which is further connected with mixed fluid receiving chamber 659 (709).
  • capillary 656 (706) comprises a sacrificial valve 662 (712).
  • Capillary channel 656 (706) is about 0.25mm deep and having a cross-sectional diameter of from about 0.5mm and that extends from about 0.2cm to about 30cm.
  • Mixing chamber 657 (707) is about 0.25mm deep and having a cross-sectional diameter of from about 0.75mm, and is positioned from about 0.2cm to about 30cm from the center of rotation.
  • Capillary channel 658 (708) is about 0.5mm deep and has a cross-sectional diameter of about 5mm and that extends from about 0.2cm to about 30cm.
  • Capillary channel 656 (706) and capillary channel 658 (708) may be offset in their connection with the mixing chamber as previously described in Example 5 , or the capillaries can be positioned in the mixing chamber at any convenient position, and Coriolis forces have be relied upon to facilitate mixing.
  • Capillary 658 (708) is fluidly connected with mixed fluid receiving chamber 659 (709).
  • Mixed fluid receiving chamber 659 (709) is about 0.75mm deep and having a cross-sectional diameter of about 5mm from about, and is positioned from about lcm to about 30cm from the center of rotation.
  • the platforms of the invention are provided containing the fluids to be mixed already in fluid reservoirs 651 and 652 (701 and 702).
  • sacrificial valves 712 be provided in capillaries 653 and 654 (703 or 704), to prevent evaporation, wetting or leakage of fluid from the reservoirs prior to use.
  • the fluids from each capillary flows past capillary junction 655 (705) and through mixing chamber 657 (707) ( Figures 18B and 18C and Figures 21B and 21C).
  • the valve prevents fluid flow into channels 653 and 654 (703 or 704).
  • fluid flow proceeds from capillary junction 655 (705) through channel 656 (706) and into mixing chamber 707 ( Figure s 18C and 21C).
  • Fluid flow within mixing chamber 657 (707) is turbulent, in contrast to fluid flow through capillary barrier 655 (705) or channel 656 (706), which is primarily laminar, so that mixing occurs predominantly in mixing chamber 657 (707). Fluid flow proceed through channel 658 (708) and the mixed fluid solution is displaced into mixed fluid receiving chamber 659 (709) ( Figures 18D and 18 E and Figures 21D and 21E).
  • EXAMPLE 7 Mixing Arrays: Second Alternative
  • FIG. 22 An additional embodiment of the microsystems platform provided by the invention and specifically designed for performing mixing of different volumes of liquid samples to form a gradient in the concentration of a species in which the two fluids differ; this embodiment is illustrated in Figure 22.
  • the a ⁇ angement of one assay a ⁇ ay 18 on a disk 11 is shown; a multiplicity of such a ⁇ ays can be advantageously a ⁇ anged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
  • the components of the mixing a ⁇ ay are shown in greater detail in Figure 23.
  • Disk embodiments of the platforms of the invention are fashioned from machined acrylic.
  • the overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device.
  • the thickness of the disc ranged from about 0.9mm to about 1.5mm.
  • the working fluid volume is about 40 ⁇ L for each fluid reservoir.
  • Fluid reservoirs 801 and 802 each containing one of a pair of liquids to be mixed, are constructed on the platform, having a depth in the platform surface from about 0.75mm and lateral dimensions of lcm, and wherein fluid reservoir 801 is designed to accommodate a volume of about 40 ⁇ L, and fluid reservoir 802 is designed to accommodate a volume of about 40 ⁇ L, wherein the volume of fluid reservoir 802 is different than the volume of fluid reservoir 801.
  • fluid reservoirs 801 and 802 are shaped so that the rate of fluid output in the two reservoirs differs between the reservoirs at a particular rotational speed, due to a change in the pressure "head" (related to the cross-sectional area of the fluid at each point in the reservoir), so that the proportion of fluid in the mixture from one of the reservoirs is at a maximum at the beginning of rotation and is at a minimum when the fluids from the reservoirs are completely mixed at the end of rotation, thus forming a gradient.
  • Gradient should be approximately 40% of left reservoir and 60% of right reservoir fluid at beginning and reversed at end; aliquoting structure of other mixing a ⁇ ay should provide a way to preserve this gradient.
  • Each fluid reservoir is fluidly connected with a capillary channel 803 or 804 to capillary junction 805.
  • Each capillary channel is about 0.5mm deep and having a cross- sectional diameter of about 0.5mm and that extends about 5cm.
  • Capillary junction 805 has a depth in the platform surface of about 0.75mm that is greater than the depth of capillaries 803 to 804.
  • capillaries 803 or 804 comprise a sacrificial valve
  • the fluid components of the mixing a ⁇ ay are also connected with air channels 810, that have dimensions of about 0.25mm deep and permit venting of air displaced by fluid movement on the platform.
  • air channels 810 that have dimensions of about 0.25mm deep and permit venting of air displaced by fluid movement on the platform.
  • capillary junctions 811 that are about
  • 0.75mm deep are present in the air channels to prevent fluid backflow into the air channel.
  • Capillary junction 805 is fluidly connected by a na ⁇ ow capillary channel 806 to mixing chamber 807, which is fluidly connected with channel 808, which is further connected with mixed fluid receiving chamber 809.
  • capillary 806 comprises a sacrificial valve 812.
  • Capillary channel 806 is about 0.5mm deep and having a cross-sectional diameter of from about 0.5mm and that extends from about 0.2cm to about 30cm.
  • Mixing chamber 807 is about 0.75mm deep and having a cross- sectional diameter of from about 0.75mm, and is positioned from about 0.2cm to about 30cm from the center of rotation.
  • Capillary channel 808 is about 0.5mm deep and has a cross-sectional diameter of about 5mm and that extends from about 0.2cm to about 30cm.
  • Capillary channel 806 and capillary channel 808 may be offset in their connection with the mixing chamber as previously described in Example 5, or they can be positioned in the mixing chamber at any convenient position, and Coriolis forces have be relied upon to facilitate mixing.
  • Capillary 808 is fluidly connected with mixed fluid receiving chamber 809.
  • Mixed fluid receiving chamber 809 is about 0.75mm deep and having a cross-sectional diameter of about 5mm from about, and is positioned from about lcm to about 30cm from the center of rotation. As illustrated in Figures 24A through 24E, in the use of this platform a volume
  • each of the fluids to be mixed is applied to the fluid reservoirs 801 and 802 ( Figure 24A). Fluid enters the each of the capillaries 803 and 804 and stops at capillary junction 805.
  • the platforms of the invention are provided containing the fluids to be mixed already in fluid reservoirs 801 and 802.
  • sacrificial valves 812 be provided in capillaries 803 and 804, to prevent evaporation, wetting or leakage of fluid from the reservoirs prior to use.
  • the fluids from each capillary flows past capillary junction 805 and through mixing chamber 807 ( Figures 24B and 24C).
  • the valve prevents fluid flow into channels 803 and 804.
  • fluid flow proceeds from capillary junction 805 through channel 806 and into mixing chamber 807 ( Figure 24D).
  • Fluid flow within mixing chamber 807 is turbulent, in contrast to fluid flow through capillary ba ⁇ ier 805 or channel 806, which is primarily laminar, so that mixing occurs predominantly in mixing chamber 807. Fluid flow proceed through channel 808 and the mixed fluid solution is displaced into mixed fluid receiving chamber 809 ( Figures 24D and 24E).
  • FIG. 25 A microsystems platform provided by the invention and specifically designed for performing immunoassay is illustrated in Figure 25.
  • the a ⁇ angement of one assay a ⁇ ay 19 on a disk 11 is shown; a multiplicity of such a ⁇ ays can be advantageously a ⁇ anged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
  • the components of the mixing a ⁇ ay are shown in greater detail in Figure 26.
  • Disk embodiments of the platforms of the invention were fashioned from machined acrylic.
  • the overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device.
  • the thickness of the disc ranged from about 0.9mm to about 1.5mm.
  • the working fluid volume for reaction was about 10 ⁇ L.
  • the components of the mixing a ⁇ ay were as follows.
  • An entry port 901 having a depth in the platform surface of about 0.75m and lateral dimensions of about 0.5cm was constructed on the platform, and designed to accommodate a volume of about 1 O ⁇ L, ranging from 2 ⁇ L to 20 ⁇ L.
  • This entry port was fluidly connected with a metering capillary 902 having cross-sectional diameter of about 0.5mm and having a depth of about 0.75mm; the length of this metering capillary was sufficient to contain a total volume of about lO ⁇ L.
  • the metering capillary 902 was fluidly connected to capillary junction 904.
  • the entry port was also constructed to be fluidly connected with an overflow capillary 903 having a cross-sectional diameter of about 0.5mm and proximal ends rounded with respect to entry port 901.
  • the overflow capillary was fluidly connected with an overflow chamber 905 having a depth in the platform surface of about 0.75mm, greater than the depth of the overflow capillary 903.
  • Each of the overflow and fluid chambers was also connected with air ports or air channels, such as 523, that have dimensions of about 0.25mm deep and permit venting of air displaced by fluid movement on the platform. Capillary junctions 524 that were about 0.75mm deep were present in the air channels to prevent fluid flow into the air channels.
  • Entry port 901 was positioned on the platform 1.3cm from the center of rotation.
  • Metering capillary 902 extends 4.2cm from entry port 9015.
  • Overflow capillary 902 extends from entry port 901 from about lcm to about 20cm.
  • the extent of the length of overflow capillary 903 was 20%> longer than metering capillary 902.
  • the position of overflow chamber 905 was from about lcm to about 20cm from the center of rotation, and the position of capillary junction 904 was 5cm from the center of rotation.
  • Capillary junction 904 was fluidly connected with capillary channel 906, which in turn was fluidly connected with incubation chamber 910.
  • Capillary channel 906 has a cross-sectional diameter of about 0.5mm and extends about lcm.
  • Capillary channel 910 has a depth in the platform surface of about 0.75mm, that was greater than the depth of capillary channel 906.
  • Capillary channel 906 was also fluidly connected with channel 909 through capillary junction 907.
  • Capillary junction 907 was constructed to prevent fluid flow of the sample backwards through the junction and into the wash buffer.
  • Channel 909 has a cross-sectional diameter of about 0.5mm and extends about 5cm.
  • Capillary junction 907 has a depth in the platform surface of about 0.75mm, greater than the depth of the channel 909 or capillary channel 906.
  • Incubation chamber 910 also contains a specific binding species, most preferably an antibody, specific for a component of the sample. This species was advantageously contained within incubation chamber 910 as a coating on the surface of the chamber, or attached to beads or other carrier within the chamber, or to a functionalized inner surface of the chamber.
  • Capillary junction 907 was further fluidly connected with wash buffer reservoir 516, having a depth in the platform surface of about 0.75mm and positioned at a distance 3.7cm from the axis of rotation.
  • Capillary junction 907 was further fluidly connected with reagent capillary 920, which was further fluidly connected with capillary junction 914, which was further fluidly connected with channel 926, and which was fluidly connected with reagent reservoir 917.
  • Reagent capillary 920 has a cross-sectional diameter of about 0.25mm and extended about lcm.
  • Capillary junction 914 has a depth in the platform surface of about 0.25mm and positioned at a distance of about 2.7cm from the axis of rotation.
  • Reagent capillary 926 has a cross-sectional diameter of about 0.25mm and extends from about 0.2cm to about 20cm.
  • Reagent reservoir 917 has a depth in the platform surface of about 0.75mm and positioned at a distance of about 2.3cm from the axis of rotation.
  • Incubation chamber 910 was fluidly connected at a point most distal to the axis of rotation to U-shaped capillary 921.
  • U-shaped capillary 921 has a cross-sectional diameter of about 0.5mm and extended about lcm. This capillary extends in a U-shape to a point that was at least as proximal to the axis of rotation than the most axis-proximal extent of incubation chamber 910.
  • Waste reservoir 915 has a depth in the platform surface of about 0.75mm and positioned at a distance of about 4.5-5.7cm from the axis of rotation.
  • sacrificial valves 922 can be positioned at the junction of capillary junction 904 and capillary channel 906, at the junction of capillary junction 907 and wash buffer capillary 908, or at the junction of reagent reservoir 918 and capillary junction 919.
  • reagent reservoir 916 and wash reservoir 915 were pre-loaded on the disk, and most preferably the disk contains sacrificial valves 922 at the junction of capillary junction 907 and wash buffer capillary 908, and at the junction of reagent reservoir 918 and capillary junction
  • wash buffer fluid flow forces the sample through
  • sacrificial valves 922 were released to permit wash buffer fluid flow.
  • Reagent buffer fluid flow forces the wash buffer through U-shaped capillary 914 and into waste reservoir 515 ( Figures 27K through 27L).
  • sacrificial valves 922 were released to permit reagent buffer fluid flow.
  • the reagent buffer contained a chromogen or other developing agent for detection of specific binding in incubation chamber 910.
  • the extent of specific immunoassay being was determined relative to the amount of chromogen produced in the assay.
  • Capture MAB was immobilized on 8- ⁇ m NC by application of 2 ⁇ L spots at a concentration of 10 mg/mL. These spots spread to circles about " in diameter. After brief incubation, the field surface of the NC was blocked in a bath of 1%> BSA in PBS, followed by extensive washing with 0.1 % BSA-PBS. The membrane was allowed to air dry prior to use. The spotted region was then cut into small squares and introduced into glass test tubes followed by 200 ⁇ L aliquots of serially diluted TSH standards in 50/50 horse serum PBS-BSA buffer. After a brief incubation to wet the NC, 10 ⁇ L of a 0.309 micron diameter blue latex suspension coated with a complimentary MAB was added and mixed. After 5 min of gentle shaking, the disk was removed, washed under a PBS-BSA stream, and allowed to dry. By visual inspection, the color intensity was seen to vary in the expected way with TSH concentration.
  • Additional antibody labeled groups such as colloidal gold, fluorescent or colored latex beads, can be used to detect antibody binding and accumulation.
  • Thyroxine A competitive assay for the monovalent thyroid hormone Thyroxine has been developed analogous to and complimenting the sandwich assay described above for TSH.
  • Antigen was immobilized on 5 ⁇ m NC by application of 1.5 ⁇ L volumes of BS A- T4 (12.5 mg/mL) followed by blocking with 1 % BSA in PBS and final washing with 0.1 % BSA in PBS.
  • Anti-thyroxine monoclonal antibody (MAB) was immobilized on
  • Antibody-coated beads were briefly incubated in test tubes with T4 in buffer over a wide hormone concentration range. Square pieces of nitrocellulose containing adsorbed capture antigen were then added to the test tubes and incubated for 10 minutes, during which time color developed on the NC. The squares were then washed with a small volume of buffer and color intensity visually noted as a function of T4 level.
  • Resistive heater elements were prepared on the platforms of the invention as follows. Portions of the surface of a microsystems platform on which a resistive heater element are desired are screen printed with a resistive ink electrically coupled with a conductive ink to conduct electricity across the resistive ink in the presence of a direct cu ⁇ ent (DC) voltage drop across the extent of the resistive ink. Both the resistive ink and the conductive ink are screened printed using methods well known in the art (Gilleo, ibid.)
  • a conductive ink pattern such as those illustrated in Figure 30, was screened onto a heat stabilized polyester sheet substrate (ICI ST505), and cured at 110- 120°C for 10 minutes.
  • a resistive ink was then screened over the cured conductive ink pattern and the composite cured for an additional 10 min at 110-120°C.
  • the ink is laid down in a layer that is about 10 microns thick, over an area at least 0.5mm x
  • the circuit materials consisted of Dupont 5028 conductive ink and Dupont 7082 resistive ink; the resistive ink comprised the solid back patterned areas and the conductive inks comprised the light, thin lined patterns on the disc.
  • Dupont 7082 resistive ink 400 Ohm per square per mil was blended with Dupont 7102 resistive ink (20 Ohm per square per mil).
  • resistive Polymer Thick Films As Heater Elements
  • the use of resistive polymer thick films as heater elements depends on the capacity of resistive polymer thick films to produce heat when an electrical potential is applied across a resistive polymer thick film element produced as described in Example 10.
  • the capacity of a resistive polymer thick film element to be used as a heater is shown in Figure 32. Circuits in the pattern shown in Figures 30 and 31 were screen- printed using Dupont 5028 silver paste as the conductive electrode combined with a 50
  • FIG. 32 is a graph of the temperature produced as a function of time and applied (DC) voltage (voltage applied was 2V, 3 V, 4V, 5 V, and 6V, read bottom to top from the voltage that produced the lowest steady-state voltage through the highest steady-state voltage). This graph shows that the maximum temperature produced increased with both time and applied voltage, and that the maximum voltage reached a steady state within between 100-150 seconds, and that the
  • Heat-activated valves include valves prepared by depositing a "wax" in a channel or capillary of the fluidics structures disclosed herein.
  • a small amount of molten wax is taken up in a preheated plastic pipette sip, and when the tip is applied to a capillary channel, the channel takes up some of the molten wax by capillary action.
  • the wax valve forms when the wax cools and solidifies in the channel or capillary.
  • a resistive heater layer is prepared on a polyester substrate and bonded to the platform substrate with tape. Heater positioning is important to ensure that the wax plug is fully melted, is capable of even flow and will not re-crystallize and clog the channel or capillary downstream of the location of the resistive heater.
  • the channels and capillaries of the invention comprising wax valves also comprise wax recrystallization chambers, illustrated in Figures 35 and 37, positioned "downstream" (that is, more proximal to the edge of the platform) from the resistive heater.
  • the resistive heating element be positioned over the entire extent of the wax valve and over at least the interior 25% of the wax recrystallization chamber.
  • Figures 36 A through 36C provide an illustration of how a platform comprising the wax valves, fluidics structures and resistive polymer thick films is prepared.
  • heater circuits are screen printed on polyester films bonded to a polycarbonate disc with double-sided tape.
  • Fluidics structures are machined onto an acrylic disc as described herein and shown in Figure 36B.
  • Wax valves positioned manually at appropriate regions of the fluidics channels and capillaries of the platforms of the invention. These two components are then bonded with double-sided tape to form the assembled disc (shown in Figure 36C).
  • Figure 37 is a more detailed and expanded view of one of the fluid handling a ⁇ ays of the invention.
  • the screen-printed lead, prepared according to Example 10 is placed on the platform so that the wax valve is completely covered with the screen printed heater element, which also extends over a portion of the wax recrystallization chamber.
  • Figure 38 illustrates how the heat-activated wax valves can be used to sequentially open the wax valves and permit fluid flow from a fluid chamber into a fluid receptacle. Aqueous dye was loaded into each of the fluid chambers shown in Figure
  • the disc was rotated at about 700 rpm, a speed above the capillary burst rpm.
  • Figure illustrates that sequential application of 10V DC across these resistive heaters is sufficient to completely melt the wax valves without reclosure of the wax valves. Also, the heating and melting of any one of these wax valves was insufficient to cause melting of any of the adjacent wax valves, which are separated on the platform by aboutl -2mm.
  • heat-recoverable polymers were used as heat activated valves.
  • heat recoverable tubing FP301H obtained from
  • the resistive heater itself was demonstrated as a sacrificial valve. It was observed that certain configurations of the resistive heaters of the invention were capable of melting the platform substrate when sufficient voltages were placed across the resistive heater. In this experiment, 15V (DC) was applied across the heaters shown in Figure 31. After less than 1 second at this voltage, a hole developed within the resistive element itself, typically at its center, having an area of about lmm 2 . These results suggested that the heater could be used as a sacrificial valve.
  • a platform comprising a chamber is prepared connected to a channel that terminates at a radius R from the center of rotation. This terminal part of the channel is then bonded to a heater circuit positioned just below the channel termination point.
  • a second fluidics disk is prepared having a channel positioned at precisely the termination point of the first channel, and is then bonded to this heater circuit so that the beginning point of this second channel is positioned just below the resistive element.
  • the resistive element will be positioned between the two channels in different layers of the platform, so that heating through the element will conjoin the two channels and permit fluid flow from one chamber to the other at a different layer of the platform.
  • the resistive heaters of the invention are also used to heat a fluid in an incubation chamber or other microfluidics component of a platform of the invention.
  • a DC voltage of 40V was applied between contacts 31 and 33 in a resistive heater produced as shown in Figure 31 , in which a fluid chamber containing about 65 ⁇ L of water was placed in thermal contact.
  • a thermistor was also placed in the fluid chamber to monitor the temperature of the water, and the platform was rotated at about 500rpm. These results are shown in Figure 39.
  • a steady-state temperature was obtained after about 200 seconds. After about 500 seconds, the heater was turned off while rotation of the platform continued.
  • screen-printed PTC inks display a larger change in resistance with temperature than other inks, these inks can be used to measure temperature on a screen- printed circuit.
  • the temperature-sensing device is screen-printed identically as a resistive heater element; however, rather than applying a voltage across the element to produce heat, the resistance of the element is sampled to determine whether resistance has increase as a result of heating.
  • the resistive heater element itself could be used both as a heater and a temperature sensing device.
  • the heater circuit is coupled to an external circuit that is switched between constant voltage and constant (small) cu ⁇ ent modes. In the constant voltage mode the element would heat up, and in the constant cu ⁇ ent mode the voltage drop would be measured and the temperature determined thereby.
  • a PTC ink is screen-printed over a portion to a resistive heater element.
  • the resistive heater and PTC ink/the ⁇ nosensor elements are joined to external circuitry using different sets of conductive leads; the external circuit delivers voltage to the resistive heater element and a small constant cu ⁇ ent to the thermosensor element.
  • the temperature of the resistive element is infe ⁇ ed. Coordination of these two circuits could be provided so that the thermosensor is used to regulate the temperature of the resistive heating element.

Abstract

This invention provides methods and apparatus for performing microanalytic and microsynthetic analyses and procedures. Specifically, the invention provides a microsystem platform for use with a micromanipulation device to manipulate the platform by rotation, thereby utilizing the centripetal force resulting from rotation of the platform to motivate fluid movement through microchannels embedded in the microplatform. The microsystem platforms of the invention are also provided having microfluidics components, resistive heating elements, temperature sensing elements, mixing structures, capillary and sacrificial valves, and methods for using these microsystems platforms for performing biological, enzymatic, immunological and chemical assays. An electronic spindle designed rotor capable of transferring electrical signals to and from the microsystem platforms of the invention is also provided.

Description

Devices and Methods for Using Centripetal Acceleration to Drive Fluid Movement in a Microfluidics System
BACKGROUND OF THE INVENTION
1. Field of the Invention This invention relates to methods and apparatus for performing microanalytic and microsynthetic analyses and procedures. In particular, the invention relates to microminiaturization of genetic, biochemical and chemical processes related to analysis, synthesis and purification. Specifically, the invention provides a microsystem platform and a micromanipulation device to manipulate the platform by rotation, thereby utilizing the centripetal forces resulting from rotation of the platform to motivate fluid movement through microchannels embedded in the microplatform. The microsystem platforms of the invention are also provided having microfluidics components, resistive heating elements, temperature sensing elements, mixing structures, capillary and sacrificial valves, and methods for using these microsystems platforms for performing biological, enzymatic, immunological and chemical assays. A slip ring designed rotor capable of transferring electrical signals to and from the microsystem platforms of the invention is also provided.
2. Summary of the Related Art In the field of medical, biological and chemical assays, mechanical and automated fluid handling systems and instruments are known in the prior art.
U.S. Patent No. 4,279,862, issued July 21, 1981 to Bertaudiere et al. disclose a centrifugal photometric analyzer.
U.S. Patent No. 4,381,291, issued April 26, 1983 to Ekins teach analytic measurement of free ligands.
U.S. Patent No. 4,515,889, issued May 7, 1985 to Klose et al. teach automated mixing and incubating reagents to perform analytical determinations. U.S. Patent No. 4,676,952, issued June 30, 1987 to Edelmann et al. teach a photometric analysis apparatus.
U.S. Patent No.4,745,072, issued May 17, 1998 to Ekins discloses immunoassay in biological fluids. U.S. Patent No. 5,061,381, issued October 29, 1991 to Burd discloses a centrifugal rotor for performing blood analyses.
U.S. Patent No. 5,122,284, issued June 16, 1992 to Braynin et al. discloses a centrifugal rotor comprising a plurality of peripheral cuvettes.
U.S. Patent No. 5,160,702, issued November 3, 1993 to Kopf-Sill and Zuk discloses rotational frequency-dependent " valves " using capillary forces and siphons, dependent on "wettablility" of liquids used to prime said siphon.
U.S. Patent No. 5,171,695, issued December 15, 1992 to Ekins discloses determination of analyte concentration using two labelling markers.
U.S. Patent No. 5,173,193, issued December 22, 1992 to Schembri discloses a centrifugal rotor for delivering a metered amount of a fluid to a receiving chamber on the rotor.
U.S. Patent No. 5,242,803, issued September 7,1993 to Burtis et al. disclose a rotor assembly for carrying out an assay.
U.S. Patent No. 5,409,665, issued April 25, 1995 to Burd discloses a cuvette filling in a centrifuge rotor.
U.S. Patent No. 5,413,009, issued July 11, 1995 to Ekins discloses a method for analyzing analytes in a liquid.
U.S. Patent No. 5,472,603, issued December 5, 1995 to Schembri discloses an analytical rotor comprising a capillary passage having an exit duct wherein capillary forces prevent fluid flow at a given rotational speed and permit flow at a higher rotational speed.
Anderson, 1968, Anal. Biochem. 28: 545-562 teach a multiple cuvette rotor for cell fractionation.
Renoe et al., 1974 Clin. Chem. 20: 955-960 teach a "minidisc" module for a centrifugal analyzer.
Burtis et al, 1975, Clin. Chem. 20: 932-941 teach a method for a dynamic introduction of liquids into a centrifugal analyzer.
Fritsche et al, 1975, Clin. Biochem. 8: 240-246 teach enzymatic analysis of blood sugar levels using a centrifugal analyzer.
Burtis et al, 1975, Clin Chem. 21_: 1225-1233 teach a multipurpose optical system for use with a centrifugal analyzer.
Hadjiioannou et al., 1976, Clin. Chem. 22: 802-805 teach automated enzymatic ethanol determination in biological fluids using a miniature centrifugal analyzer.
Lee et al, 1978, Clin. Chem. 2A: 1361-1365 teach a automated blood fractionation system. Cho et al, 1982, Clin. Chem. 28: 1956-1961 teach a multichannel electrochemical centrifugal analyzer.
Bertrand et al, 1982, Clinica Chimica Acta 119: 275-284 teach automated determination of serum 5'-nucleotidase using a centrifugal analyzer.
Schembri et al, 1992, Clin Chem. 38: 1665-1670 teach a portable whole blood analyzer.
Walters et al, 1995, Basic Medical Laboratory Technologies. 3rd ed., Delmar Publishers: Boston teach a variety of automated medical laboratory analytic techniques.
Recently, microanalytical devices for performing select reaction pathways have been developed. U.S. Patent No. 5,006,749, issued April 9, 1991 to White disclose methods apparatus for using ultrasonic energy to move microminiature elements.
U.S. Patent No. 5,252,294, issued October 12, 1993 to Kroy et al. teach a micromechanical structure for performing certain chemical microanalyses.
U.S. Patent 5,304,487, issued April 19, 1994 to Wilding et al. teach fluid handling on microscale analytical devices.
U.S. Patent 5,368,704, issued November 29, 1994 to Madou et al. teach microelectrochemical valves.
International Application, Publication No. W093/22053, published 11 November 1993 to University of Pennsylvania disclose microfabricated detection structures. International Application, Publication No. W093/22058, published 11 November
1993 to University of Pennsylvania disclose microfabricated structures for performing polynucleotide amplification.
Columbus et al, 1987, Clin. Chem. _33: 1531-1537 teach fluid management of biological fluids.
Ekins et al, 1994 Ann. Biol. Clin. 50: 337-353 teach a multianalytic microspot immunoassay.
Wilding et al, 1994, Clin. Chem. 40: 43-47 disclose manipulation of fluids on straight channels micromachined into silicon.
One drawback in the prior art microanalytical methods and apparati has been the difficulty in designing systems for moving fluids on microchips through channels and reservoirs having diameters in the 10-100μm range. Micro fluidic systems require precise and accurate control of fluid flow and valving to control chemical reactions and analyte detection. Conventional pumping and valving mechanisms have been difficult to incorporate into microscale structures due to inherent conflicts-of-scale. These conflicts of scale arise in part due to the fact that molecular interactions arising out of mechanical components of such components, which are negligible in large
(macroscopic) scale devices, become very significant for devices built on a microscopic scale.
Systems that use centripetal force to effect fluid movement in microstructures address the need for a pumping mechanism to effect fluid flow, but cannot alone solve these scale-related drawbacks of conventional fluidics reduced to microfluidics scale.
There remains a need for a simple, flexible, reliable, rapid and economical microanalytic and microsynthetic reaction platform for performing biological, biochemical and chemical analyses and syntheses that can move fluids within the structural components of a microsystems platform. Such a platform should be able to move nanoliter-to microliter amounts of fluid, including reagents and reactants, at rapid rates to effect the proper mixing of reaction components, removal of reaction side products, and isolation of desired reaction products and intermediates. There remains a need in the art for centripetally-motivated microfluidics platforms capable of precise and accurate control of flow and metering of fluids in both microchip-based and centrifugal microplatform-based technologies. SUMMARY OF THE INVENTION
This invention provides microsystems platforms as disclosed in co-owned and co-pending U.S. Serial No. 08/761,063, filed December 5, 1996 and incorporated by reference herein. Specifically, this invention provides microfluidics components, resistive heating elements, temperature sensing elements, mixing structures, capillary and sacrificial valves, and methods for using these microsystems platforms for performing biological, enzymatic, immunological and chemical assays.
It is an advantage of the centrifugal rotors and microsystems platforms of the invention that an imprecise amount of a fluid comprising a biological sample can be applied to the rotor or platform and a precise volumetric amount of the biological sample is delivered to a fluid reservoir comprising a reaction vessel or other component of the rotor of platform for performing chemical, biochemical, immunological or other analyses. It is an advantage of the centrifugal rotors and microsystems platforms of the invention that metering of said precise amount of a biological fluid sample, for example, a drop of blood, is provided as an intrinsic property of the metering capillary channel of the rotor or platform, thereby avoiding variability introduced by centripetal metering of the sample into a reaction reservoir. It is a further advantage of the centrifugal rotors and microsystems platforms of the invention that an operator can avoid having to precisely measure an amount of a fluid comprising a biological sample for application to the rotor or microsystem platform, thereby permitting end-users, including consumers, having a lower level of sophistication to use a medically diagnostic or other embodiment of the rotor or microsystem platform of the invention.
It is an advantage of the centrifugal rotors and microsystems platforms of the invention that fluid movement into and out of fluid reservoirs on the rotor or platform is precisely determined by displacement of a first fluid, such as biological sample, from a fluid reservoir by a second fluid contained in a second reservoir on the rotor or platform. It is also an advantage of the centrifugal rotors and microsystems platforms of the invention that approximately complete replacement of the volumetric capacity of a first reservoir can be achieved by using fluid displacement as disclosed herein, thereby providing for maximum recovery of a first fluid sample upon displacement by a second fluid, or maximum delivery and replacement of the first fluid by the second fluid. This aspect of the invention is advantageous for providing sequential chemical or biochemical reaction steps wherein mixing of the reagents is not desired.
Specific preferred embodiments of the present invention will become evident from the following more detailed description of certain preferred embodiments and the claims.
DESCRIPTION OF THE DRAWINGS
Figures 1 through 3 illustrate the microfluidics array of the microsystem platform described in Example 1. Figures 4 through 6 illustrate the microfluidics array of the microsystem platform described in Example 2.
Figures 7 through 9 illustrate the microfluidics array of the microsystem platform described in Example 3.
Figures 10 through 12 illustrate the microfluidics array of the microsystem platform described in Example 4.
Figures 13 through 1511ustrate the microfluidics array of the microsystem platform described in Example 5.
Figures 16 through 18 illustrate the microfluidics array of the microsystem platform described in Example 6. Figures 19 through 21 illustrate the microfluidics array of the microsystem platform described in Example 7.
Figures 22 through 24 illustrate the microfluidics array of the microsystem platform described in Example 8.
Figures 25 through 27 illustrate the microfluidics array of the microsystem platform described in Example 9.
Figure 28 illustrates a microfluidics array of a multiplicity of mixing chambers of the microsystem platform described in Example 8.
Figure 29 illustrates the electronic spindle of the invention.
Figure 30 illustrates screen-printing of a resistive heater element as described in
Example 10. Figure 31 illustrates screen-printing of a resistive heater element as described in Example 10.
Figure 32 is a graph showing the time dependence of temperature produced at a variety of voltages using a resistive heater element as described in Example 10. Figure 33 is a graph showing the voltage dependence of temperature produced at a variety of voltages using a resistive heater element as described in Example 10.
Figure 34 is a graph showing the distance dependence on heating using a resistive heater element as described in Example 10.
Figure 35 illustrates screen-printing of a resistive heater element associated with wax valves as described in Example 11.
Figure 36 illustrates screen-printing of a resistive heater element associated with wax valves as described in Example 11.
Figure 37 illustrates screen-printing of a resistive heater element associated with wax valves as described in Example 11. Figure 38 illustrates the use of screen-printed resistive heater elements to melt wax valves and control fluid flow in a microfluidics array as described in Example 11.
Figure 39 is a graph showing that temperature can be cycled using a screen- printed resistive heater element as described in Example 14.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
The present invention provides centrifugal rotors and microsystems platforms for providing centripetally-motivated fluid micromanipulation.
For the purposes of this invention, the term "sample" will be understood to encompass any fluid, solution or mixture, either isolated or detected as a constituent of a more complex mixture, or synthesized from precursor species.
For the purposes of this invention, the term "in fluid communication" or "fluidly connected" is intended to define components that are operably interconnected to allow fluid flow between components. In preferred embodiments, the platform comprises a rotatable platform, more preferably a disk, whereby fluid movement on the disk is motivated by centripetal force upon rotation of the disk. For the purposes of this invention, the term "a centripetally motivated fluid micromanipulation apparatus" is intended to include analytical centrifuges and rotors, microscale centrifugal separation apparati, and most particularly the microsystems platforms and disk handling apparati of International Application No. WO97/21090. For the purposes of this invention, the term "microsystems platform" is intended to include centripetally-motivated microfluidics arrays as disclosed in International Application No. WO97/21090.
For the purposes of this invention, the terms "capillary", "microcapillary" and "microchannel" will be understood to be interchangeable and to be constructed of either wetting or non-wetting materials where appropriate.
For the purposes of this invention, the term "fluid chamber" will be understood to mean a defined volume on a rotor or microsystems platform of the invention comprising a fluid.
For the purposes of this invention, the term "entry port" will be understood to mean a defined volume on a rotor or microsystems platform of the invention comprising a means for applying a fluid to the rotor or platform.
For the purposes of this invention, the term "capillary junction" will be understood to mean a junction of two components wherein one or both of the lateral dimensions of the junction are larger than the corresponding dimensions the capillary. In wetting or wettable systems, the such junctions are where capillary valving occurs, because fluid flow through the capillaries is stopped at such junctions. In non-wetting or non-wettable junctions, the exit from the chamber or reservoir is where the capillary junction occurs. In general, it will be understood that capillary junctions are formed when the dimensions of the components change from a small diameter (such as a capillary) to a larger diameter (such as a chamber) in wetting systems, in contrast to non- wettable systems, where capillary junctions form when the dimensions of the components change from a larger diameter (such as a chamber) to a small diameter (such as a capillary).
For the purposes of this invention, the term "biological sample" or "biological fluid sample" will be understood to mean any biologically-derived analytical sample, including but not limited to blood, plasma, serum, lymph, saliva, tears, cerebrospinal fluid, urine, sweat, plant and vegetable extracts, semen, and ascites fluid.
For the purposes of this invention, the term "air displacement channels " will be understood to include ports in the surface of the platform that are contiguous with the components (such as chambers and reservoirs) on the platform, and that comprise vents and microchannels that permit displacement of air from components of the platforms and rotors by fluid movement.
For the purposes of this invention, the term "capillary action" will be understood to mean fluid flow in the absence of rotational motion or centripetal force applied to a fluid on a rotor or platform of the invention. For the purposes of this invention, the term "capillary micro valve" will be understood to mean a capillary comprising a capillary junction whereby fluid flow is impeded and can be motivated by the application of pressure on a fluid, typically by centripetal force created by rotation of the rotor or platform of the invention.
The microplatforms of the invention (preferably and hereinafter collectively referred to as "disks"; for the purposes of this invention, the terms "microplatform",
"microsystems platform" and "disk" are considered to be interchangeable), are provided to comprise one or a multiplicity of microsynthetic or microanalytic systems. Such microsynthetic or microanalytic systems in turn comprise combinations of related components as described in further detail herein that are operably interconnected to allow fluid flow between components upon rotation of the disk. These components can be fabricated as described below either integral to the disk or as modules attached to, placed upon, in contact with or embedded in the disk. The invention also comprises a micromanipulation device for manipulating the disks of the invention, wherein the disk is rotated within the device to provide centripetal force to effect fluid flow on the disk. Accordingly, the device provides means for rotating the disk at a controlled rotational velocity, for stopping and starting disk rotation, and advantageously for changing the direction of rotation of the disk. Both electromechanical means and control means, as further described herein, are provided as components of the devices of the invention. User interface means (such as a keypad and a display) are also provided, as further described in International Application WO97/21090.
Fluid (including reagents, samples and other liquid components) movement is controlled by centripetal acceleration due to rotation of the platform. The magnitude of centripetal acceleration required for fluid to flow at a rate and under a pressure appropriate for a particular microsystem is determined by factors including but not limited to the effective radius of the platform, the position angle of the structures on the platform with respect to the direction of rotation and the speed of rotation of the platform.
The capillary junctions and microvalves of the invention are based on the use of rotationally-induced fluid pressure to overcome capillary forces. Fluids which completely or partially wet the material of the microchannels (or reservoirs, reaction chambers, detection chambers, etc.) which contain them experience a resistance to flow when moving from a microchannel of narrow cross-section to one of larger cross- section, while those fluids which do not wet these materials resist flowing from microchannels (or reservoirs, reaction chambers, detection chambers, etc.) of large cross- section to those with smaller cross-section. This capillary pressure varies inversely with the sizes of the two microchannels (or reservoirs, reaction chambers, detection chambers, etc., or combinations thereof), the surface tension of the fluid, and the contact angle of the fluid on the material of the microchannels (or reservoirs, reaction chambers, detection chambers, etc.) . Generally, the details of the cross-sectional shape are not important, but the dependence on cross-sectional dimension results in microchannels of dimension less than 500μm exhibit significant capillary pressure. By varying the intersection shapes, materials and cross-sectional areas of the components of the microsystems platform of the invention, "valve" are fashioned that require the application of a particular pressure on the fluid to induce fluid flow. This pressure is applied in the disks of the invention by rotation of the disk (which has been shown above to vary with the square of the rotational frequency, with the radial position and with the extent of the fluid in the radial direction). By varying capillary valve cross-sectional dimensions as well as the position and extent along the radial direction of the fluid handling components of the microsystem platforms of the invention, capillary valves are formed to release fluid flow in a rotation-dependent manner, using rotation rates of from lOOrpm to several thousand rpm. This arrangement allows complex, multistep fluid processes to be carried out using a pre-determined, mono tonic increase in rotational rate. The theoretical principles underlying the use of capillary junctions and microvalves are disclosed in International Patent Application, Publication No. WO97/ .
The instant invention provides microsystems platforms comprising microfluidics components, heating elements, temperature sensing elements, capillary valves, sacrificial valves and a rotor design for transmitting electrical signals to and from the microsystems platforms of the invention. The invention provides fluidics components for capillary metering of precise amounts of a volume of a fluid sample from the application of a less precise volume of a fluid sample at an entry port on the microsystem platform. These embodiments of the invention provide for delivery of precise amounts of a sample such as a biological fluid sample without requiring a high degree of precision or accuracy by the operator or end-user in applying the fluid to the platform, and is advantageous in embodiments of the microsystems platforms of the invention that are used by consumers and other relatively unsophisticated users. The invention also provides laminar flow- dependent replacement of a fluid in a first chamber by a second displacement fluid in a second chamber on the platform. These embodiments of the invention provide approximately complete replacement of a fluid in one chamber on the platform with fluid from another, and thereby provide means for practicing sequential chemical reactions and other sequential processes on the platform under conditions wherein mixing of the two fluids is disadvantageous. The invention also provides turbulent flow mixing components, which permit thorough mixing of different fluid components on the platform. In particular, the invention provides mixing chambers fluidly connected with fluid reservoirs containing equal amounts of two or more different fluids or unequal amounts of two or more different fluids. In addition, the invention provides fluid reservoirs fluidly connected with mixing chamber of the invention and shaped to determine the relative rate of flow of each of the different fluids into the mixing chamber. In these embodiments, gradients of two fluids differing in viscosity, solute concentration or concentration of suspended particulates can be produced using the mixing chambers of the invention. Such gradients can be transferred to reservoirs on the platform for further analytical manipulations, and can form the basis for controlled testing of concentration-dependent effects of various catalysts, drugs, toxins or other biological or chemical agents. Platforms of the invention such as disks and the components comprising such platforms are advantageously provided having a variety of composition and surface coatings appropriate for a particular application. Platform composition will be a function of structural requirements, manufacturing processes, and reagent compatibility/chemical resistance properties. Specifically, platforms are provided that are made from inorganic crystalline or amorphous materials, e.g. silicon, silica, quartz, inert metals, or from organic materials such as plastics, for example, poly(methyl methacrylate) (PMMA), acetonitrile-butadiene-styrene (ABS), polycarbonate, polyethylene, polystyrene, polyolefms, polypropylene and metallocene. These may be used with unmodified or modified surfaces. Surface properties of these materials may be modified for specific applications. Surface modification can be achieved by silanization, ion implantation and chemical treatment with inert-gas plasmas (i.e., gases through which electrical currents are passed to create ionization). Also provided by the invention are platforms made of composites or combinations of these materials,/or example, platforms manufactures of a plastic material having embedded therein an optically transparent glass surface comprising for example the detection chamber of the platform. Microplatform disks of the invention are preferably fabricated from thermoplastics such as teflon, polyethylene, polypropylene, methylmethacrylates and polycarbonates, among others, due to their ease of molding, stamping and milling. Alternatively, the disks can be made of silica, glass, quartz or inert metal. A fluid handling system is built by sequential application of one or more of these materials laid down in stepwise fashion onto the thermoplastic substrate. Disks of the invention are fabricated with an injection molded, optically-clear base layer having optical pits in the manner of a conventional compact disk (CD). The disk is a round, polycarbonate disk 120mm in diameter and 100 pm thick. The optical pits provide means for encoding instrument control programming, user interface information, graphics and sound specific to the application and driver configuration. The driver configuration depends on whether the micromanipulation device is a handheld, benchtop or floor model, and also on the details of external communication and other specifics of the hardware configuration. This layer is then overlaid with a reflective surface, with appropriate windows for external detectors, specifically optical detectors, being left clear on the disk. Other layers of polycarbonate of varying thickness are laid down on the disk in the form of channels, reservoirs, reaction chambers and other structures, including provisions on the disk for valves and other control elements. These layers can be pre-fabricated and cut with the appropriate geometries for a given application and assembled on the disk. Layers comprising materials other than polycarbonate can also be incorporated into the disk. The composition of the layers on the disk depend in large part on the specific application and the requirements of chemical compatibility with the reagents to be used with the disk. Electrical layers can be incorporated in disks requiring electric circuits, such as electrophoresis applications and electrically-controlled valves. Control devices, such as integrated circuits, laser diodes, photodiodes and resistive networks that can form selective heating areas or flexible logic structures can be incorporated into appropriately wired recesses, either by direct fabrication of modular installation onto the disk. Reagents that can be stored dry can be introduced into appropriate open chambers by spraying into reservoirs using means similar to inkjet printing heads, and then dried on the disk. A top layer comprising access ports and air vents, ports or shafts is then applied. Liquid reagents are then injected into the appropriate reservoirs, followed by application of a protective cover layer comprising a thin plastic film.
The platforms of the invention are preferably provided with a multiplicity of components, either fabricated directly onto the platform, or placed on the platform as prefabricated modules. In addition to the integral components, certain devices and elements can be located external to the platform, optimally positioned on a device of the invention in relation to the platform, or placed in contact with the platform either while rotating or when at rest. Components optimally comprising the platforms of the invention or a controlling device in combination therewith include detection chambers, reservoirs, valving mechanisms, detectors, sensors, temperature control elements, filters, mixing elements, and control systems.
This invention provides microsystems platforms comprising the following components.
1. Microfluidics Components
The platforms of the invention are provided comprising microfluidics handling structures in fluidic contract with one another. In preferred embodiments, fluidic contact is provided by capillary or microchannels comprising the surface of the platforms of the invention. MicroChannel sizes are optimally determined by specific applications and by the amount of delivery rates required for each particular embodiment of the platforms and methods of the invention. MicroChannel sizes can range from 0.1 m to a value close to the 1mm thickness of the platform. MicroChannel shapes can be trapezoid, circular or other geometric shapes as required. Microchannels preferably are embedded in a platform having a thickness of about 0.1 to 100mm, wherein the cross-sectional dimension of the microchannels across the thickness dimension of the platform is less than 500μm and from 1 to 90 percent of said cross-sectional dimension of the platform.
In these embodiments, which is based on the use of rotationally-induced fluid pressure to overcome capillary forces, it is recognized that fluid flow is dependent on the orientation of the surfaces of the components. Fluids which completely or partially wet the material of the microchannels, reservoirs, detection chambers, etc. (i.e., the components) of the platforms of the invention which contain them experience a resistance to flow when moving from a component of narrow cross-section to one of larger cross-section, while those fluids which do not wet these materials resist flowing from components of the platforms of the invention of large cross-section to those with smaller cross-section. This capillary pressure varies inversely with the sizes of the two components, or combinations thereof, the surface tension of the fluid, and the contact angle of the fluid on the material of the components . Generally, the details of the cross- sectional shape are not important, but the dependence on cross-sectional dimension results in microchannels of dimension less than 500μm exhibit significant capillary pressure. By varying the intersection shapes, materials and cross-sectional areas of the components of the platform of the invention, "valves" are fashioned that require the application of a particular pressure on the fluid to induce fluid flow. This pressure is applied in the disks of the invention by rotation of the disk (which has been shown above to vary with the square of the rotational frequency, with the radial position and with the extent of the fluid in the radial direction). By varying capillary valve cross-sectional dimensions as well as the position and extent along the radial direction of the fluid handling components of the platforms of the invention, capillary valves are formed to release fluid flow in a rotation-dependent manner, using rotation rates of from lOOrpm to several thousand rpm. This arrangement allows complex, multistep fluid processes to be carried out using a pre-determined, monotonic increase in rotational rate.
A first example of the microfluidics arrays provided by this invention is shown in Figures 1 through 3. A microsystems platform is provided by the invention that is specifically designed for performing two-step assays. These Figures illustrate an array advantageously used for any two-step analytic process; an antibiotic detection assay is illustrated herein.
Such a microsystem platform is illustrated in Figure 1. In the Figure, the arrangement of one assay array 12 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
The components of the antibiotic assay array are shown in greater detail in Figure 2. It will be understood by a comparison of Figures 1 and 2 that the center of the platform 11 is at the top of Figure 2, and the edge or lateral extent of the platform is at the bottom of Figure 2, illustrated by a curved line. Rotation of the antibiotic array on platform disks of the invention can be in either direction, although rotation in a consistent, particular direction is preferred. Disc embodiments of the platforms of the invention were fashioned from machined acrylic and injection-molded polycarbonate. The overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device. The thickness of the disc ranged from about 0.9mm to about 1.5mm. The working fluid volume for reaction with reagents was about l-150μL.
The components of the antibiotic array are as follows. An entry port 201 having a depth in the platform surface ranging from about 0.25mm to about 6mm and lateral dimensions of from about 0.2cm to about 2cm is constructed on the platform, and designed to accommodate a volume of about 5-100μL. This entry port is fluidly connected with an array of metering capillaries 202 having a square cross-sectional diameter ranging from about 0.02mm to about 1mm and proximal ends rounded with respect to entry port 201; the length of this metering capillary array was sufficient to contain a total volume of about 10-5 OμL. The entry port is also constructed to be fluidly connected with an overflow capillary 203 having a cross-sectional diameter of from about 0.02mm to about 1mm and proximal ends rounded with respect to entry port 201. The overflow capillary is fluidly connected with an overflow chamber 205 having a depth in the platform surface ranging from about 0.02mm to about 5mm, provided that the depth of the overflow chamber 205 is greater than the depth of the overflow capillary
203. Metering capillary 202 is fluidly connected to fluid chamber 204 having a depth in the platform surface ranging from about 0.02mm to about 1mm, provided that the depth of the fluid chamber 204 is greater than the depth of the metering capillary 202. Each of the overflow and fluid chambers is also connected with airports or air channels, such as 211 , that have dimensions ranging from about 0.02mm to about 1mm and permit venting of air displaced by fluid movement on the platform. A capillary junction 212 that is about 0.75mm to 1mm deep is present in the air channel to prevent fluid flow into the air channel. For the purposes of the description of these microfluidics structures, a "capillary junction" is defined in a hydrophilic support material as a pocket, depression or chamber that has a greater depth (vertically within the platform) and/ or a greater width (horizontally within the platform) that the fluid channel to which it is fluidly connected. For liquid with a contact angle less than 90° (such as aqueous solutions on platforms made with most plastics, glass and silica), flow is impeded as the channel cross-section increases at the interface of the capillary junction. The force hindering flow is produced by capillary pressure, which is inversely proportional to the cross sectional dimensions of the channel and directly proportional to the surface tension of the liquid, multiplied by the cosine of the contact angle of the fluid in contact with the material comprising the channel. The factors relating to capillarity in microchannels according to this invention have been discussed in co-owned and co-pending U.S. patent application, Serial No. 08/910,726, filed August 12, 1997, incorporated by reference in its entirety herein.
Entry port 201 is positioned on the platform from 1cm to 20cm from the center of rotation. Metering capillary 202 extends from entry port 201 from about 0.2cm to about 20cm. The extent of the length of overflow capillary 203 is at least about 20% greater than the extent of the length of metering capillary 202. The position of fluid chamber 204 is from about 0.5cm to about 10cm from the center of rotation, and the position of overflow chamber 205 is thus from about 1.5cm to about 11.5cm from the axis of rotation.
The fluid chamber 204 acts as a capillary barrier that prevents fluid flow from metering capillary 202 at a first, non-zero rotational speed f, sufficient to permit fluid flow comprising overflow from the entry port 201 through overflow capillary 203 and into overflow chamber 205. The capillary boundary of fluid chamber 204 is constructed to be overcome at a second rotational speed f2 (where f2 > f,). Fluid chamber 204 is fluidly connected to capillary 206 that is from about 0.02mm to about 1mm deep and has a cross-sectional diameter ranging from about 0.02mm to about 1mm. Capillary 206 extends from about 0.1cm to about 20cm from fluid chamber 204 and is connected to holding chamber 207. Holding chamber 207 has a depth in the platform surface from about 0.02mm to about 1mm and greater than the depth of capillary 206. Filling of fluid chamber 204 is motivated by centripetal acceleration and is accompanied by fluid flow through capillary 206 into holding chamber 207. Holding chamber 207 is fluidly connected by way of capillary 208, having a cross-sectional diameter of from about
0.1mm to about 1mm and extending from about 0.2cm to about 10cm from holding chamber 207 and is further connected with read chamber 210. Read chamber 210 has a depth in the platform surface of from about 0.02mm to about 5mm. For optical detection methods, read chamber 210 is of sufficient optical quality to prevent excessive scattering for a diffusely-reflecting medium such as milk. For transparent samples, read chamber 210 comprises a reflector, for example. This may be a diffuse reflector on the side of the chamber opposite to that upon which light is incident or a mirror- like surface. In the case of a diffuse reflector such as paint, a porous material, or any other material which causes diffuse reflection because of its rough surface or porous nature, light incident at a given angle is emitted over a hemisphere with a well-known cosine dependence on angle. In this way the detector may be aligned at an angle not equal to the incident angle, so that specular reflection from the plastic window of the chamber does not enter the detector. Alternatively, the mirror-like surface reflects light at an angle equal to the incident angle. In certain embodiments, a sacrificial valve 213 (as described herein below) is placed as shown in the channel 209. The use of a platform as disclosed in Figures 1 and 2 is illustrated in Figures 3 A through 3J. In the use of this platform an imprecise volume (ranging from l-150μL of fluid) of a fluid is applied to the entry port 201 (Figure 3 A). In embodiments of the platform comprising air displacement channels, the fluid will wick into air channel 211 and be stopped by capillary junction 212. Fluid also wicks into metering capillary 202 and overflow capillary 203. Fluid flows through the metering capillary 202 and overflow capillary 203 at zero rotational speed until the fluid reaches capillary junctions at the junction between metering capillary 202 and fluid chamber 204 and overflow capillary 203 and overflow chamber 205 (Figures 3B and 3C). Metering capillary 202 is constructed to define a precise volume from about l-150μL of fluid between entry port 201 and the capillary junction at fluid chamber 204, which is designed to be at least the amount of the fluid placed by the user in entry port 201.
After sample loading by a user and filling of metering capillary 202 and overflow capillary 203 at zero rotational speed (which can be performed on the disc separately or with then disc engaged with a spindle of a centrifugal device), the platform is spun at a first rotational speed f,, ranging from 100-400rpm; the exact value is dependent on the position of the capillary junction components on the platform. For example, for an entry port 201 having a depth of 0.6mm, metering capillary 202 having dimensions of 0.5mm x 0.5mm in cross-section and a capillary junction between metering capillary 202 and fluid chamber 204 positioned about 2.2-3.8cm from the center of rotation, and overflow capillary 203 having dimensions of 0.5mm x 0.5mm in cross-section and a capillary junction between overflow capillary 203 and overflow chamber 205 positioned about 5.4cm from the center of rotation, this first rotational speed f, is equal to about 175rpm for either water or milk. Due to the greater distance of the end of overflow capillary 203 from the center of rotation than the end of metering capillary 202, fluid flows through overflow capillary 203 into overflow chamber 205 and does not flow from metering capillary 202 into fluid chamber 204 at rotational speed {,. The platform is spun until all excess fluid is evacuated from entry port 201 and into overflow chamber 205, except the fluid contained in metering capillary 202 (Figure 3D).
At a second rotational speed f2, that is greater than the first rotational speed fl5 typically in the range of 400-520rpm, the precise amount of fluid contained in metering capillary 202 is delivered into fluid chamber 204 (Figures 3E through 3H) For example, for an entry port 201 having a depth of 0.6mm, metering capillary 202 having dimensions of 0.5mm x 0.5mm in cross-section and a capillary junction between metering capillary 202 and fluid chamber 204 positioned about of 2.2-3.8cm from the center of rotation, this second rotational speed is equal to 400rpm for either water or milk. Fluid movement into fluid chamber 204 is accompanied by filling of capillary 206 and holding chamber 207.
In embodiments comprising a sacrificial valve 213 in-line with capillary 208 at position 209 shown in Figure 2, release of the sacrificial valve results in fluid flow into read chamber 210. Sacrificial valves as described above are preferably made of a fungible material that can be removed from the fluid flow path. In preferred embodiments, said sacrificial valves are wax valves and are removed from the fluid flow path by heating, using any of a variety of heating means including infrared illumination and most preferably by activation of heating elements on or embedded in the platform surface as described below. In said embodiments, fluid flow is achieved at rotational speed f2 with removal of the sacrificial valve.
In embodiments of the platforms of the invention comprising antibiotic arrays as described herein and not containing a sacrificial valve at position 209, capillary 208 preferably fills along with filling of holding chamber 207 until the fluid reaches a capillary junction 209 at the junction between capillary 208 and read chamber 210; in such embodiments, the capillary junction has a depth of from about 0.15mm to about lmm. At a third rotational speed f3, that is greater than the second rotational speed f2, typically in the range of >520rpm, the fluid contained in holding chamber 207 is delivered into read chamber 210 (Figures 31 and 3J). For example, for a capillary junction 209 having a depth of 0.75mm, and capillary 208 having dimensions of 0.25mm x 0.25mm in cross-section and the capillary junction positioned from about 3.8cm from the center of rotation, this third rotational speed is equal to 500-800rpm for either water or milk. More generally, the third rotational speed is proportional to l/ (R0Uter - Rιnner) x
{(Router x RinnerV 2 x the diameter of the capillary}, where Route- is the dimension of the capillary radius at the outer edge and Rmner is the capillary radius at the inner edge, relative to the center of rotation. Thus, for a capillary of square cross-section 0.25mm and a capillary junction 0.75mm deep, values of the product {(Router - Rιnner) x {(Router x Rmner)/ x the diameter of the capillary} ranging from 0.89 to 0.35 yield a third rotational speed of from about 500-800 rpm.
An example of the chemistries of the types of two-step chemical analyses that can be performed using this microfluidics array are illustrated by the antibiotic detection assay. This antibiotic detection assay is performed using the platform design as follows. This chemistry is based on the selective poisoning of an enzyme, carboxypeptidase, by beta lactam antibiotics. Carboxypeptidase is present in a dried form with sugars , buffers or other stabilizers in the holding chamber 207 and a metered amount of milk or other fluid sample solution is introduced into the holding chamber as described above. The milk or other fluid sample solubilizes the enzyme, and the milk/enzyme mixture is incubated at 47° for 3-5 minutes to allow any beta lactam antibiotics present to bind to the carboxypeptidase. The enzyme catalyses cleavage of a D-alanine residue from L-
Lysine-D-Ala-D-Ala, and catalysis is inhibited in a concentration-dependent manner by beta-lactam antibiotics present in the sample. In preferred embodiments, this temperature is achieve using a resistive heater element as described below; a description of this resistive heater element is not specifically included in this description of the platform of the invention here in order to focus attention on the disclosed microfluidics structures. It will be understood that any analytic protocol using the microfluidics structures disclosed herein that requires elevated temperatures (i.e., greater than room temperature) advantageously includes resistive heaters as disclosed herein, or other heating elements that are specifically encompassed by the platforms of the invention. In addition, the holding chamber 207 contains dried reagent comprising a peptide having a D-amino acid at its carboxyl terminus; an example of such a peptide is L- Lysine-D-Ala-D-Ala. Preferably, this peptide is also contained in holding chamber 207 in dried form, and is reconstituted by introduction of milk or other fluid into the chamber as described above. It is an advantage of the microfluidics arrays of the invention that a precisely metered amount of fluid is introduced into holding chamber 207, permitting a standardized amount of carboxypeptidase, peptide substrate and buffers, stabilizers, etc. be contained in the holding chamber.
In the second step of the assay, D-amino acid oxidase (DAAO), flavine adenine dinucleotide (FAD), horse radish peroxidase (HRP), and a chromogen such as syringalazine (4-hydroxy-3,5-dimethoxygenzaldehyde azine) or ortho-dianisidine (ODA) is contained in dried form in read chamber 210. Displacement of the fluid sample from holding chamber 207 to read chamber 210 results in reconstitution of these reagents, which produce a colored product proportional with the amount of D-amino acid present in the fluid sample after incubation with carboxypeptidase and D-amino acid containing peptide. The D-alanine residue produced by degradation of the peptide substrate by carboxypeptidase in holding chamber 207 is degraded into pyruvate in the presence of
DAAO and FAD; the reaction also reduces FAD to FADH2. FADH2 combines with oxygen in the fluid sample to produce hydrogen peroxide and be reoxidized to FADH2 to FAD. The hydrogen peroxide then acts on a chromogen such as syringalazine in the presence of horse radish peroxidase, and the previously transparent chromogen becomes colored. This reaction scheme is illustrated as follows:
(carboxypeptidase + H20) (DAAO + FAD)
L-Lysine-D-Ala-D-Ala > D-Ala >
47°C 47°C
(02) (HRP + chromogen) Pyruvate + FADH2 H202+ FAD > oxidized chromogen
The extent of chromogen production is detected in the read chamber, and related to the presence of antibiotics in the sample by comparison with samples tested in the absence of antibiotic. Most preferably, a standard curve relating the decrease in chromogen production and the amount of antibiotic in the sample is prepared and used to determine the amount of antibiotic in an unknown test sample.
The buffers and reagents used for these chemistries on the platforms of the invention are constituted with appropriate buffers, stabilizers, preservatives, salts, cofactors, adjuvants and other necessary components of the reactions performed on the platform. An alternative embodiment of the two-step assay microfluidics array is shown in Figures 4 through 6, again exemplified for an antibiotic assay disk of the invention. It will be understood that, as in Example 1 , in Figure 4, the arrangement of one assay array 13 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
Disk embodiments of the platforms of the invention were fashioned from machined acrylic. The overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device. The thickness of the disc ranged from about 0.9mm to about 1.5mm. The working fluid volume for reaction with reagents was about 25-150μL.
The components of this antibiotic array are as follows. An entry port 301 having a depth in the platform surface of from about 0.25mm to about 5mm and lateral dimensions of from about 0.2cm to about 2cm is constructed on the platform, and designed to accommodate a volume of about l-100μL. This entry port is fluidly connected with one or a multiplicity of entry capillaries 302 having a square cross- sectional diameter ranging from about 0.02mm to about 1mm and having a depth of 0.1 to 1mm equal to the depth of the entry port, and proximal ends rounded with respect to entry port 301. The length of this metering capillary array was sufficient to contain a total volume of about 20μL . The entry capillaries 302 are fluidly connected to fluid chamber 303 having a depth in the platform surface of from about 0.02mm to about lmm, wherein the depth is greater than the depth of the entry capillary 302. Each of the fluid chambers of this aspect of the invention is also connected with air ports or air channels, such as 311, that have dimensions ranging from about 0.02mm to about 5mm that permit venting of air displaced by fluid movement on the platform. A capillary junction 312 that is about 0.75mm deep is present in the air channel to prevent fluid flow into the air channel.
The fluid chamber 303 is also constructed to be fluidly connected with an overflow capillary 304 having a cross-sectional diameter of about 0.02mm to about 0.75mm and proximal ends rounded with respect to fluid chamber 304. The overflow capillary is fluidly connected with an overflow chamber 306 having a depth in the platform surface of from about 0.02mm to about lmm, greater than the depth of the overflow capillary 304.
Entry port 301 is positioned on the platform from 1cm to 20cm from the center of rotation. Entry capillaries 302 extends from entry port 301 from about 0.5cm to about 10cm. The position of a first fluid chamber 303 is from about 0.5cm to about 10cm from the center of rotation.
The first fluid chamber 303 acts as a capillary barrier that prevents fluid flow from entry capillary 302 at zero rotational speed. Movement of fluid from entry port 301 through entry capillaries 302 and into the first fluid chamber 303 is achieved by rotation at a first, non-zero rotational speed f , . Displacement of fluid into the first fluid chamber
303 is accompanied by fluid filling of channel 305 that is fluidly connected with the first fluid chamber 303 and is positioned at the most radially distal point of the first fluid chamber. Channel 305 is fluidly connected with a second fluid chamber 307 and forms a capillary boundary between channel 305 and chamber 307. This capillary boundary is constructed to be overcome at second rotational speed f2 (where f2 > fj). First fluid chamber 303 is also fluidly connected to overflow capillary 304 that is from about 0.05mm to about lmm deep and has a cross-sectional diameter of from about 0.05mm to about lmm and extends from about 0.2cm to about 20cm. Overflow capillary 304 is connected to overflow chamber 306 that has a depth in the platform surface equal to that of overflow capillary 304, so that there is no capillary boundary between overflow capillary 304 and overflow chamber 306. Overflow capillary 304 is positioned in the first fluid chamber 303 at a point radially less distant from entry port 301 than channel 305, thereby defining a volume in the fluid chamber between the position of the overflow capillary 304 and the most radially distant extent of the said first fluid chamber.
Second fluid chamber 307 is further fluidly connected through channel 308 to a small pocket or capillary junction 309, having a depth in the platform surface of from about 0.1mm to about 5mm and positioned about 0.2 to 20cm from the axis of rotation. Channel 308 has a cross-sectional diameter ranging from about 0.02mm to about lmm and extends from about 0.2cm to about 10cm, and further extends to a third fluid chamber 310. Third fluid chamber 310 has a depth in the platform surface of from about 0.1mm to about 5mm, that is greater than the depth of capillary 308. Air recirculation channels 311 that have dimensions of from about 0.02mm to about lmm provide pathways for air displaced by fluid movement, while capillary junctions 312 that are about 0.75mm deep prevent fluid from entering the air channels. In some embodiments of the device a sacrificial valve 313 is placed as shown in the channel 309. In certain embodiments, a valve 314 is placed in channel 305 to control fluid movement from the first fluid chamber 303 to the second fluid chamber 307.
The use of this platform is illustrated in Figures 6 A through 6J. An imprecise volume (ranging from l-150μL of fluid) of a fluid is applied to the entry port 301 (Figure 6A). Fluid wicks into entry capillary 302 and stops at the capillary junction between entry capillary 302 and the first fluid chamber 303 (Figures 6B and 6C). Fluid flows through the entry capillary B and into the first fluid chamber 303 at a first rotational speed fl5 ranging from 100-500rpm; the exact value is dependent on the position of the components on the platform (Figures 6D and 6E). For example, for an entry port 301 having a depth of 0.75mm, entry capillary 302 having dimensions of
0.25mm x 0.5mm in cross-section and a length of 0.5-lcm from the center of rotation, this first rotational speed f, is equal to about 250rpm for either water or milk. The fluid further enters capillary channel 305, stopping at the capillary junction with the second fluid chamber 307. As rotation continues, the fluid continues to fill the first fluid chamber 303, overflow capillary 304 fills (Figure 6F), and excess fluid fills overflow chamber 306 until the level of fluid in the first fluid chamber 303 falls below the position of overflow capillary 304 (Figure 6G).
At a second rotational speed f2, that is greater than the first rotational speed fl5 typically in the range of 100-1 OOOrpm, the capillary junction between channel 305 and the second fluid chamber 307 is overcome, and fluid remaining in the first fluid chamber
303 is delivered into the second fluid chamber 307 (Figures 6H and 61). For example, for channel 305 having dimensions of 0.25mm x 0.5mm in cross-section and a length of 2.5-3.3cm from the center of rotation, this second rotational speed is equal to 280rpm for either water or milk. In an alternative embodiment, a sacrificial valve 314 is placed at the junction of channel 305 and the second fluid chamber 308, which sacrificial valve is released to permit fluid flow through channel 305 and into the second fluid chamber 308. In such embodiments, fluid flow can be achieved at either f, or f2 rotational velocity.
In embodiments comprising a sacrificial valve 313 in-line with capillary 308 at position 309 shown in Figure 5, release of the sacrificial valve results in fluid flow into the third fluid chamber 310. Sacrificial valves are as described above are preferably made of a fungible material that can be removed from the fluid flow path. In preferred embodiments, said sacrificial valves are wax valves and are removed from the fluid flow path by heating, using any of a variety of heating means including infrared illumination and most preferably by activation of heating elements on or embedded in the platform surface as described below. In said embodiments, fluid flow is achieved at rotational speed f2 with removal of the sacrificial valve.
In embodiments of the platforms of the invention comprising antibiotic arrays as described herein and not containing a sacrificial valve at position 310, capillary 309 preferably fills along with filling of the second chamber 308 until the fluid reaches a capillary junction 309 at the junction between capillary 308 and the third fluid chamber
310; in such embodiments, the capillary junction has a depth of about 0.75mm (ranging from about 0.25mm to about lmm). At a third rotational speed f,, that is greater than the second rotational speed f2, typically in the range of >500rpm, the fluid contained in the second chamber 308 is delivered into the third fluid chamber 310 (Figures 6H through 6K). For example, capillary 309 having dimensions of 0.25mm x 0.25mm in cross- section and a length of 3.36-3.7cm from the center of rotation, this third rotational speed is equal to 400rpm for either water or milk.
This embodiment of the platforms of the invention can be used for any two-step analytical assay. Using the antibiotic assay described above as an example, the second fluid chamber 308 contains a reagent, such as carboxypeptidase, and its substrate, for example L-lysine-D-alanine-D-alanine, and incubation is performed therein to produce D-Ala. The remaining reagents are placed in the third fluid chamber 310, and the reaction including color development advantageously proceeds in situ whereby chamber 310 is a read chamber. The extent of chromogen production is detected in the read chamber, and related to the presence of antibiotics in the sample by comparison with samples tested in the absence of antibiotic. Most preferably, a standard curve relating the decrease in chromogen production and the amount of antibiotic in the sample is prepared and used to determine the amount of antibiotic in an unknown test sample.
The invention also provides microfluidics arrays for separating the fluid component from a particulate suspension. An example of such a parti culate suspension is blood, where red and white blood cells are suspended in plasma. Thus, this aspect of the microfluidics embodiments of the invention is illustrated using separation of blood plasma from whole blood.
A microsystems platform provided by the invention and specifically designed for separating vertebrate blood cells and components is illustrated in Figures 7 through 9. In Figure 7, the arrangement of one assay array 14 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
The components of the blood separation array are shown in greater detail in Figure 8. It will be understood by a comparison of Figures 7 and 8 that the center of the platform 11 is at the top of Figure 8, and the edge or lateral extent of the platform is at the bottom of Figure 8, illustrated by a curved line. Rotation of the blood separation array on platform disks of the invention can be in either direction, although rotation in a consistent, particular direction is preferred. Disk embodiments of the platforms of the invention were fashioned from machined acrylic. The overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device. The thickness of the disc ranged from about 0.9mm to about 1.5mm. The working fluid volume was about l-50μL.
The components of the blood separation array are as follows. An entry port 401 having a depth in the platform surface ranging from about 0.1mm to about 5mm and lateral dimensions of about 0.1 to about 2cm is constructed on the platform, and designed to accommodate a volume of about 5 to about 50μL. This entry port is fluidly connected to an entry capillary 402, having a cross-sectional diameter of from about 0.02mm to about lmm and a depth of about 0.5 to lmm. The length of this entry capillary was sufficient to contain a total volume of from about 1 to about 15μL. Entry capillary 402 is further fluidly connected to a separation column 403 having a cross- sectional diameter of from about 0.1mm to about 2mm, a depth of from about 0.25mm to about lmm, and a length sufficient to contain a total volume of 10 to about 20μL. This separation column is also fluidly connected with a passage 411 to overflow chamber 404. Passage 411 has a cross-sectional diameter ranging from about 0.5mm to about 2mm, a depth of about 0.25mm to about lmm, and a length of 0.5mm to about 5mm). Overflow chamber 404 has a depth of from about 0.25-lmm.
A small capillary exit 406 is also fluidly connected with separation chamber 403, having a cross-sectional diameter of from about 0.05mm to about0.25mm, a depth of about 0.025mm to about 0.125mm, and a length about 0.25mm to about 5mm. This capillary is arranged to traverse a direction radially more proximal to the axis of rotation than the insertion point with separation column 403. This small capillary 406 terminates in a capillary junction 407 that is fluidly connected with capillary 408, extending in a radial direction to decant chamber 405. A sacrificial valve 413 is positioned in small capillary 406 at the juncture with the capillary junction. Capillary 408 has a cross- sectional diameter ranging from about 0.05mm to about lmm, a depth of about 0.05mm to about lmm, and a length of from about lmm to about 100mm. This capillary is arranged in a radially outward direction between capillary junction 407 and decant chamber 405. Passage 411 is positioned on separation column 403 to be significantly more proximal to the axis of rotation than the insertion point of small capillary 406.
Air displacement channels 409 that have dimensions of from about 0.02mm to about lmm permit venting of air displaced by fluid movement on the platform.
Capillary junctions 410 that are about 0.75mm deep are present in the air channels to prevent fluid flow into the air channels.
The use of this platform is illustrated in Figures 9A through 9H for separating plasma from whole blood. An imprecise volume (ranging from l-150μL of fluid) of blood is applied to the entry port 401 (Figure 9A). Blood enters the entry capillary 402 by capillary action, and stops at the capillary junction between entry capillary 402 and the separation chamber 403 (Figures 9B and 9C). At a first rotational speed f,, ranging from 100-300rpm (the exact value is dependent on the position of the components on the platform), blood flows from the entry capillary 402 into separation chamber 403 (Figure 9D). Blood continues to fill separation column 403 until blood reaches the position of passage 411, whereupon excess blood flows through passage 411 and into overflow chamber 404 (Figures 4E and 4F). Advantageously, small channel 406 has dimensions that prevent wicking of blood into the channel as blood flows past the insertion point of small channel 406 into separation column 403.
As shown in Figure 9F, after sufficient time of rotation at the first non-zero rotational speed f„ the excess blood has been transferred into overflow chamber 404 and the separation column 403 is filled with blood to the position of passage 411. Rotation at a second rotational speed f2, that is greater than the first rotational speed f,, typically in the range of 1000-5000rpm, blood components are separated into red blood cell, white blood cell (i.e., "buffy coat"), and plasma fractions (Figure 9G). Advantageous dimensions of small capillary 406 permit fluid flow of the plasma fraction through capillary 406 that is stopped at capillary junction 407. Fluid flow of plasma into decant chamber 405 results from fluid flow overcoming the capillary barrier 407 by rotation at a third rotational speed f3, that is greater than the second rotational speed f2, typically in the range of >1000-5000rpm (Figure 9H). An alternative embodiment of the fluid separation platform is also provided by the invention, again illustrated by the separation of plasma from whole blood. This embodiment of the blood separation microfluidics array is shown in Figures 10 through 12. It will be understood that, as above, in Figure 10, the arrangement of one separation array 15 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform. Disk embodiments of the platforms of the invention are fashioned from machined acrylic. The overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device. The thickness of the disc ranged from about 0.9mm to about 1.5mm. The working fluid volume was about 5-50μL.
The components of this separation array are as follows. An entry port 501 having a depth in the platform surface of about 0.1mm to about lmm and lateral dimensions of about 0.1cm to about 2cm is constructed on the platform, and designed to accommodate a volume of about 5 to about 50μL. This entry port is fluidly connected with a first array of metering capillaries 502 and a second array of metering capillaries
503, wherein each of the capillaries has a cross-sectional diameter of from about 0.02mm to about lmm. The length of the second metering capillary array 503 is longer than that of the first metering capillary array 502. The first metering capillary array 502 is fluidly connected with a ballast chamber 507, having a depth in the platform surface ranging radially from about 0.1mm to about 5mm and greater than the depth of the first metering capillary array 502, wherein the first metering capillary array 502 forms a capillary junction between the array and the ballast chamber. The second capillary array 503 is fluidly connected with capillary junction 506.
The entry port is also constructed to be fluidly connected with an overflow capillary 504 having a cross-sectional diameter of about 0.02mm to about lmm and proximal ends rounded with respect to entry port 501. The overflow capillary is fluidly connected with an overflow chamber 505 having a depth in the platform surface of from about 0. lmm to about 5mm, greater than the depth of the overflow capillary 504. Each of the overflow and fluid chambers is also connected with airports or air channels, such as 514, that have dimensions ranging from about 0.1mm to about lmm and that permit venting of air displaced by fluid movement on the platform. Capillary junctions 516 that are about 0.75mm deep are present in the air channels to prevent fluid flow into the air channels.
Entry port 501 is positioned on the platform from 0.5cm to 20cm from the center of rotation. Metering capillary array 502 extends from about 0.6cm to about 10cm from entry port 501. Metering capillary array 503 extends about 0.5cm to about 10cm from entry port 501. The length of metering capillary array 503 is about at least about 20% longer than metering capillary array 502, and the extent of the length of overflow capillary 504 is at least about 20% greater than the extent of the length of either the first metering capillary array 502 or the second metering capillary array 503. The position of ballast chamber 507 is about 1cm to about 10cm from the center of rotation, the position of capillary junction 506 is about 1.5 to 15cm from the center of rotation, and the position of overflow chamber 505 is thus about 2.5 to about 20cm from the axis of rotation.
The ballast chamber 507 acts as a capillary barrier that prevents fluid flow from the first metering capillary array 502 at a first, non-zero rotational speed fj sufficient to permit fluid flow comprising excess blood overflow from the entry port 501 through overflow capillary 504 and into overflow chamber 505. Capillary junction 506 is a capillary barrier that prevents fluid flow from the second metering capillary array 503 at said first, non-zero rotational speed f, sufficient to permit fluid flow comprising excess blood overflow from the entry port 501 through overflow capillary 504 and into overflow chamber 505. These capillary boundaries are constructed to be overcome at a second rotational speed f2 (where f2 > f,).
Ballast chamber 508 is fluidly connected to capillary 510 that is from about 0.02mm to about lmm deep and has a cross-sectional diameter of about 0.02mm to about lmm and that extends from about 0.1cm to about 5cm. Capillary 510 is connected to capillary junction 511. Alternatively, capillary 510 is fluidly connected with a sacrificial valve 515. Sacrificial valves as described below are preferably made of a fungible material that can be removed from the fluid flow path. In preferred embodiments, said sacrificial valves are wax valves and are removed from the fluid flow path by heating, using any of a variety of heating means including infrared illumination and most preferably by activation of heating elements on or embedded in the platform surface. In said embodiments, fluid flow is achieved at rotational speed f2 with removal of the sacrificial valve. Sacrificial valve 515 or capillary junction 511 are further fluidly connected with channel 512 which is from about 0.1mm to about lmm deep and has a cross-sectional diameter of about 0.1mm to about lmm. Channel 512 extends about 0.1cm to about 20cm and is fluidly connected with separation chamber 509 at a point most distal from the axis of rotation.
Second metering capillary array 503 is fluidly connected with capillary junction 506, which is overcome at a rotational speed f2 > f,. Capillary junction 506 is further fluidly connected to channel 508, which is further fluidly connected to separation chamber 509. Channel 508 is from about 0.02mm to about lmm deep and has a cross- sectional diameter of from about 0.02mm to about lmm. Channel 508 extends from about t _cm to about jkcm. Separation chamber 509 is from about 0.2mm to about 5mm deep and has a cross-sectional dimension ranging from about lmm to about 20mm, and is positioned from about 10mm to about 100mm from the center of rotation. Separation chamber 509 is fluidly connected with decant channel 517 at a point close to the chamber's most axis-proximal extent. Decant channel 517 is about 0.02mm to about lmm deep and has a cross-sectional diameter ranging from about 0.02mm to about lmm. Decant channel 517 extends from about 4.3cm to about 5cm and is fluidly connected with decant chamber 514. Decant chamber 514 is from about 0.2mm to about 2mm deep and has a cross-sectional diameter of from about lmm to about 10mm. Decant chamber 514 is positioned about 5.2cm from the center of rotation..
The use of this embodiment of the microfluidics separation arrays of the invention is illustrated in Figures 12A through 12J. An imprecise volume (ranging from l-150μL of fluid) of blood is applied to the entry port 501 (Figure 12A). Blood enters the each of the metering capillary arrays 502 and 503 and stops at the capillary junction between metering capillary array 502 and ballast chamber 507 and between metering capillary 503 and capillary junction 506 (Figures 12B and 12C). Blood also enters and fills overflow capillary 504, stopping at the capillary junction with overflow chamber 505.
At a first rotational speed f,, ranging from 100-500rpm (the exact value is dependent on the position of the components on the platform), blood flows from the entry port 501 through overflow capillary 504 and into overflow chamber 505 (Figures 12D and 12E). At a second rotational speed f2, that is greater than the first rotational speed f,, typically in the range of 300-800rpm, the capillary junction between the first metering capillary array 502 and ballast chamber 508 is overcome, and blood from the first metering capillary array fills ballast chamber 508 (Figure 12F). Similarly, at second rotational speed f2, capillary junction 506 is overcome, and blood from second metering capillary array 503 enters separation chamber 509 (Figure 12F). Advantageously, the volume of blood in second metering capillary array 503 is insufficient to fill separation chamber 509 to the level of insertion of decant channel 517. By rotation at a third rotational speed f3, that is greater than the second rotational speed f2, typically in the range of 1000-5000rpm, blood components in separation chamber 509 are separated into red blood cell, white blood cell (i.e., "buffy coat"), and plasma fractions (Figures 12G and 12H). Separation of blood components is not achieved in ballast chamber 507, due to its position on the platform, and the capillary junction 511or sacrificial valve 515 are not overcome at third rotational speed f3.
Advantageously, the separated plasma does not extend to decant capillary 517. Release of sacrificial valve 517, or rotation at a fourth rotational speed f4, that is greater than the third rotational speed f3, and typically in the range of 1000-5000rpm, result in flow of blood from ballast chamber 508 through channel 512 and into separation chamber 509 at the "bottom" or most axis-distal extent of the separation chamber (Figure 121). This results in filling of the separation chamber to a position equal to the insertion point of decant channel 517 (Figure 12J). Plasma flow through decant channel 517 and into decant chamber 514 in an amount equal to the amount of blood contained in ballast chamber 508. Decant channel 517 is advantageously provided with dimensions that retard passage of unfractionated blood, or plasma contaminated with greater than 0.1-1% of blood cells found in whole blood.
The invention also provides mixing chambers and arrays of mixing chambers for mixing two or more fluids that differ in viscosity, solute concentration or concentration of suspended particulates. A first embodiment of microfluidics platform comprising the mixing chambers and arrays of the invention is shown in Figures 13- 15 for mixing equal volumes of different liquids. In Figure 13, the arrangement of one assay array 15 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform.
The components of the mixing array are shown in greater detail in Figure 14. It will be understood by a comparison of Figures 13 and 14 that the center of the platform
11 is at the top of Figure 14, and the edge or lateral extent of the platform is at the bottom of Figure 14, illustrated by a curved line. Rotation of the mixing array on platform disks of the invention can be in either direction, although rotation in a consistent, particular direction is preferred. Disk embodiments of the platforms of the invention were fashioned from machined acrylic. The overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device. The thickness of the disc ranged from about 0.9mm to about 1.5mm. The working fluid volume was about 50μL.
The components of the mixing array are as follows. Entry ports 601 having a depth in the platform surface ranging from about 0.1mm to about lmm and lateral dimensions of from about 1cm to about 5cm are constructed on the platform, and designed to accommodate a volume of about 5-50μL. Each entry port is fluidly connected with one of a paired array of metering capillaries 602 having a square cross- sectional diameter of from about 0.1mm to about 0.5mm and proximal ends rounded with respect to entry ports 601 ; the length of each metering capillary array was sufficient to contain a total volume of about 25μL. Metering capillaries 602 are fluidly connected to a curved capillary barrier 603 having a depth in the platform surface of about 0.25mm to about lmm that is greater than the depth of metering capillaries 602. The capillary barrier 603 and other fluid components of the mixing array are also connected with air channels 608, that have dimensions ranging from about 0.25mm to about lmm and that permit venting of air displaced by fluid movement on the platform. In addition, capillary junctions 609 that are about 0.75mm deep are present in the air channels to prevent fluid backflow into the air channel.
Capillary barrier 603 is fluidly connected by a naπow capillary channel 604 to mixing chamber 605, which is fluidly connected with channel 610, which is further connected with mixed fluid receiving chamber 606. Alternatively, capillary 604 comprises a sacrificial valve 612. Sacrificial valves used in this embodiment of the invention are as described below. Capillary channel 604 ranges from about 0.1mm to about lmm in depth and has a cross-sectional diameter of from about 0.1mm to about lmm). Capillary channel 604 extends from about 0.1 to about 10 cm. Mixing chamber 605 is about 0. lmm to about lmm deep and has a cross-sectional diameter of from about
0.5mm to about 5mm), is positioned about 1cm to about 30cm from the center of rotation. Capillary channel 610 ranges from about 0.1mm to about lmm deep and has a cross-sectional diameter of from about 0.1 mm to about 1 mm). Capillary channel 610 extends from about 0.2cm to about 30cm. In an advantageous embodiment, mixing chamber 605 is constructed such that the insertion point of capillary channel 604 and the insertion point of capillary channel 610 are offset at opposite ends of the mixing chamber. As a consequence, fluid flowing through capillary channel 604 is forced to encounter the opposite wall of mixing chamber 605 before fluid flow can proceed through capillary channel 610. This results in the creation of turbulence in the mixed laminar fluid stream in capillary channel 604 caused by the conjoint flow of fluid from the first and second metering channels without appreciable mixing. The turbulence created by the structure of mixing chamber 605 is sufficient to disrupt laminar flow and cause fluid mixing in the chamber prior to continued fluid flow through capillary channel 610 and into mixed fluid receiving chamber 606. Alternatively, the positions of the capillaries 604 and 610 can be at any convenient position in mixing chamber 605, wherein the Coriolis forces of fluid flow are sufficient to disrupt laminar flow and provide turbulence that results in efficient mixing.
Mixed fluid receiving chamber 606 is about 0.1mm to about 5mm deep, has a cross-sectional diameter of from about lmm to about 20mm, and is positioned from about 1cm to about 30cm from the center of rotation. The use of this embodiment of the invention for mixing equal volumes of fluid
(ranging from l-150μL is illustrated in Figures 15A through 15D. Equal volumes of each fluid to be mixed is applied to the entry ports 601 (Figure 15 A). Fluid enters the each of the metering capillary arrays 602 and stops at capillary barrier 603.
At a first rotational speed f,, ranging from 50-500rpm (the exact value is dependent on the position of the components on the platform), the fluids from each capillary aπay flows into and fills the capillary barrier 603 (Figure 15B). In embodiments comprising a sacrificial valve 612, the valve prevents fluid flow into channel 604; otherwise, fluid flow proceeds into channel 604 at rotational speed f1# Upon release of sacrificial valve 612, fluid flow proceeds from capillary junction 603 through channel 604 and into mixing chamber 605 (Figure 15C). Fluid flow within mixing chamber 605 is turbulent, in contrast to fluid flow through capillary barrier 603 or channel 604, which is primarily laminar, so that mixing occurs predominantly in mixing chamber 605. Fluid flow proceeds through channel 610 and the mixed fluid solution is displaced into mixed fluid receiving chamber 606 (Figure 15D). The invention also provides mixing arrays for mixing unequal volumes of fluid.
One example of such an additional embodiment of the microsystems platform provided by the invention and specifically designed for performing mixing of unequal volumes of different liquid samples is illustrated in Figures 19 through 21. In Figure 19, the arrangement of one assay array 17 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform. The components of the mixing array are shown in greater detail in Figure 20. It will be understood by a comparison of Figures 19 and 20 that the center of the platform 11 is at the top of Figure 20, and the edge or lateral extent of the platform is at the bottom of Figure 20, illustrated by a curved line. Rotation of the mixing array on platform disks of the invention can be in either direction, although rotation in a consistent, particular direction is preferred. Disk embodiments of the platforms of the invention are fashioned from machined acrylic. The overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device. The thickness of the disc ranged from about 0.9mm to about 1.5mm. The working fluid volume is about 2-200μL.
The components of this mixing array are as follows. Fluid reservoirs 701 and 702, each containing one of a pair of liquids to be mixed, are constructed on the platform, having a depth in the platform surface ranging from about 0.1mm to about 5mm and lateral dimensions of from about 0.2cm to about 10cm). In this embodiment, fluid reservoir 701 is designed to accommodate a volume of about 1 to about 500μL, and fluid reservoir 702 is designed to accommodate a volume of about ranging from about 1 to about 500μL, wherein the volume of fluid reservoir 702 is less than the volume of fluid reservoir 701. In particular and in addition, the viscosity of the fluid in the fluid reservoirs may differ, so that mixing produces a mixed fluid of intermediate viscosity. Also, in this embodiment the concentration of solute of suspended particulate may differ between the fluids. Each fluid reservoir is fluidly connected with a capillary channel 703 or 704 to capillary junction 705. Each capillary channel is from about 0.0.2mm to about lmm deep, has a cross-sectional diameter ranging from about 0.1mm to about lmm, and extends about 2cm to about 100cm. Capillary junction 705 has a depth in the platform surface ranging from about 0.02mm to about 1 mm that is greater than the depth of capillaries 703 to 704. Alternatively, capillaries 703 or 704 comprise a sacrificial valve 712. Sacrificial valves used in this embodiment of the invention are as described below. Use of said sacrificial valves can be used in addition to or in place of capillary junction 705. The fluid components of the mixing array are also connected with air channels
710, that have dimensions ranging from about 0. lmm to about lmm and permit venting of air displaced by fluid movement on the platform. In addition, capillary junctions 711 that are about 0.75mm deep are present in the air channels to prevent fluid backflow into the air channel.
Capillary junction 705 is fluidly connected by a narrow capillary channel 706 to mixing chamber 707, which is fluidly connected with channel 708, which is further connected with mixed fluid receiving chamber 709. Alternatively, capillary 706 comprises a sacrificial valve 712, as described below. Capillary channel 706 ranges from about 0.1mm to about lmm, has a cross-sectional diameter of from about 0.1mm to about lmm and extends from about 0.2cm to about 30cm. Mixing chamber 707 is from about 0.1mm to about lmm deep, has a cross-sectional diameter ranging from about 0. lmm to about lmm, and is positioned from about 0.2cm to about 30cm from the center of rotation. Capillary channel 708 ranges from about 0.1mm to about lmm, has a cross-sectional diameter ranging from about lmm to about 20mm and extends from about 0.2cm to about 30cm. Capillary channel 706 and capillary channel 708 are advantageously offset in their connection with the mixing chamber as described above, or are positioned at any convenient position in the mixing chamber for those embodiments relying on Coriolis forces to create mixing.
Capillary 708 is fluidly connected with mixed fluid receiving chamber 709. Mixed fluid receiving chamber 709 is about 0.1mm to about lmm, has a cross-sectional diameter of about lmm to about 20mm, and is positioned from about 1cm to about 30cm from the center of rotation.
The use of this embodiment of the microfluidics components of the invention is illustrated in Figures 21A through 21E. A volume (ranging from l-150μL of fluid) of each of the fluids to be mixed is applied to the fluid reservoirs 701 and 702 (Figure 21 A). Fluid enters the each of the capillaries 703 and 704 and stops at capillary junction
705. Alternatively, the platforms of the invention are provided containing the fluids to be mixed already in fluid reservoirs 701 and 702. In these embodiments, it is preferred that sacrificial valves 712 be provided in capillaries 703 and 704, to prevent evaporation, wetting or leakage of fluid from the reservoirs prior to use. At a first rotational speed f,, ranging from 100-lOOOrpm (the exact value is dependent on the position of the components on the platform), the fluids from each capillary flows past capillary junction 605 and through mixing chamber 707 (Figures 21B and 21C). In embodiments comprising a sacrificial valve 712, the valve prevents fluid flow into channels 703 and 704. Upon release of sacrificial valve 712, fluid flow proceeds from capillary junction 705 through channel 706 and into mixing chamber 707 (Figure 21C). Fluid flow within mixing chamber 707 is turbulent, in contrast to fluid flow through capillary barrier 705 or channel 706, which is primarily laminar, so that mixing occurs predominantly in mixing chamber 707. Fluid flow proceed through channel 708 and the mixed fluid solution is displaced into mixed fluid receiving chamber 709 (Figures 2 ID and 2 IE). In another embodiment of the mixing chamber arrays of the invention are provided platforms capable of forming a gradient of two or more liquids that differ in viscosity, solute concentration or concentration of suspended particulates. Such an additional embodiment of the microsystems platform provided by the invention and specifically designed for performing mixing of different volumes of liquid samples to form a gradient in the concentration of a species in which the two fluids differ is illustrated in Figures 22-24. In Figure 22, the arrangement of one assay array 18 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform. The components of the mixing array are shown in greater detail in Figure 23. It will be understood by a comparison of Figures 22 and 23 that the center of the platform 11 is at the top of Figure 23, and the edge or lateral extent of the platform is at the bottom of Figure 23, illustrated by a curved line. Rotation of the mixing array on platform disks of the invention can be in either direction, although rotation in a consistent, particular direction is preferred. Disk embodiments of the platforms of the invention are fashioned from machined acrylic. The overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device. The thickness of the disc ranged from about 0.9mm to about 1.5mm. The working fluid volume is about 40μL. The components of the mixing array are as follows. Fluid reservoirs 801 and
802, each containing one of a pair of liquids to be mixed, are constructed on the platform, having a depth in the platform surface from about 0.1mm to about 5mm and lateral dimensions ranging from about 1cm to about 10cm. Fluid reservoir 801 is designed to accommodate a volume of from about 1 to about 500μL, and fluid reservoir 802 is designed to accommodate a volume of about 1 to about 500μL, wherein the shape of fluid reservoir 802 is different than the shape of fluid reservoir 801. In particular and in addition, fluid reservoirs 801 and 802 are shaped so that the rate of fluid output in the two reservoirs differs between the reservoirs at a particular rotational speed, due to a change in the pressure "head" (related to the cross-sectional area of the fluid at each point in the reservoir), so that the proportion of fluid in the mixture from one of the reservoirs is at a maximum at the beginning of rotation and is at a minimum when the fluids from the reservoirs are completely mixed at the end of rotation, thus forming a gradient. Gradients produced according to this aspect of the invention can consist of salt gradients, including sodium chloride and cesium chloride or sulfate gradients, gradients of low molecular weight species such as sucrose, synthetic polymer gradients such as Ficoll or Hypaque, or gradients of a drug, toxin, enzyme substrate or other small molecule of interest.
Each fluid reservoir is fluidly connected with a capillary channel 803 or 804 to capillary junction 805. Each capillary channel ranges from about 0.1mm to about lmm deep, has a cross-sectional diameter of from about 0.1mm to about lmm and extends about 2cm to about 100cm. Capillary junction 805 has a depth in the platform surface of from about 0.1mm to about lmm that is greater than the depth of capillaries 803 to 804. Alternatively, capillaries 803 or 804 comprise a sacrificial valve 812, as described below. Use of said sacrificial valves can be used in addition to or in place of capillary junction 805. The fluid components of the mixing array are also connected with air channels
810, that have dimensions of from about 0. lmm to about lmm and permit venting of air displaced by fluid movement on the platform. In addition, capillary junctions 811 that are about 0.75mm deep are present in the air channels to prevent fluid backflow into the air channel. Capillary junction 805 is fluidly connected by a narrow capillary channel 806 to mixing chamber 807, which is fluidly connected with channel 808, which is further connected with mixed fluid receiving chamber 809. Alternatively, capillary 806 comprises a sacrificial valve 812. Capillary channel 806 is from about 0.1mm to about lmm, has a cross-sectional diameter of from about 0.1mm to about lmm and extends from about 0.2cm to about 30cm. Mixing chamber 807 is from about 0.1mm to about lmm, has a cross-sectional diameter of from about 0.1mm to about lmm, and is positioned from about 0.2cm to about 30cm from the center of rotation. Capillary channel 808 is from about 0. lmm to about lmm, has a cross-sectional diameter of about lmm to about 20mm and extends from about 0.2cm to about 30cm. Capillary channel 806 and capillary channel 808 are advantageously offset in their connection with the mixing chamber as described above, or are positioned at any convenient position in the mixing chamber for those embodiments relying on Coriolis forces to create mixing.
Capillary 808 is fluidly connected with mixed fluid receiving chamber 809. Mixed fluid receiving chamber 809 is about 0.75mm deep (ranging from about 0.1mm to about lmm) and having a cross-sectional diameter of about 5mm (ranging from about lmm to about 20mm) from about, and is positioned from about 1 cm to about 30cm from the center of rotation.
Use of this microfluidics platform to produce a gradient as described herein is illustrated in Figures 24A through 24E. A volume (ranging from l-150μL of fluid) of each of the fluids to be mixed is applied to the fluid reservoirs 801 and 802 (Figure 24 A). Fluid enters the each of the capillaries 803 and 804 and stops at capillary junction
805. Alternatively, the platforms of the invention are provided containing the fluids to be mixed already in fluid reservoirs 801 and 802. In these embodiments, it is preferred that sacrificial valves 812 be provided in capillaries 803 and 804, to prevent evaporation, wetting or leakage of fluid from the reservoirs prior to use. At a first rotational speed f,, ranging from 100-lOOOrpm (the exact value is dependent on the position of the components on the platform), the fluids from each capillary flows past capillary junction 805 and through mixing chamber 807 (Figures 24B and 24C). In embodiments comprising a sacrificial valve 812, the valve prevents fluid flow into channels 803 and 804. Upon release of sacrificial valve 812, fluid flow proceeds from capillary j unction 805 through channel 806 and into mixing chamber 807
(Figure 24D). Fluid flow within mixing chamber 807 is turbulent, in contrast to fluid flow through capillary barrier 805 or channel 806, which is primarily laminar, so that mixing occurs predominantly in mixing chamber 807. Fluid flow proceed through channel 808 and the mixed fluid solution is displaced into mixed fluid receiving chamber 809 (Figures 24D and 24E). In addition to the embodiments described herein, the invention provides microfluidics arrays comprising a multiplicity of mixing chambers fluidly connected in series to one another. Such an arrangement is illustrated in Figure 28. In particular, mixing cascades as shown in Figure 28 are useful for mixing liquids of dramatically different volumes or viscosities, such as mixing a low volume, high viscosity liquid with a high volume, low viscosity liquid. In these embodiments, the mixing array comprises a multiplicity of mixing chambers aπayed radially across the platform surface, each mixing chamber having an inlet capillary extending radially from a position more proximal to the center of rotation than the mixing chamber, and an outlet capillary extending radially to a position more distal to the center of rotation than the mixing chamber. In an advantageous embodiment of this array, the capillaries are connected with the mixing chamber so that their positions in the mixing chamber are offset from one another, wherein fluid flow from the inlet capillary impinges on a wall of the mixing chamber at a position other than the position occupied by the outlet capillary, thereby producing turbulent flow within the mixing chamber that mixes the fluids. Alternatively, the capillaries can be positioned in the mixing chamber at any convenient position, and Coriolis forces have be relied upon to facilitate mixing. In either type of array, the inlet capillary of the first mixing chamber is fluidly connected with the fluid reservoirs (either directly or through a capillary junction), and the inlet capillary of the other mixing chamber in the array is the outlet channel of the mixing chamber immediately more proximal to the center of rotation; similarly, the outlet capillary of each mixing chamber is the inlet capillary of the mixing chamber immediately more distal to the center of rotation, and wherein the outlet capillary of the mixing chamber positioned the most distal from the center of rotation is fluidly connected with a mixed fluid receiving chamber. Thus, the fluid is repetitively mixed in a multiplicity of mixing chambers arrayed more and more distally from the center of rotation, ending with a reservoir or chamber sufficient in volume to accommodate the volume of the mixed fluid. The dimensions of the capillaries, fluid reservoirs, mixing chambers and mixed fluid receiving chamber in these arrays are as described above.
Figure 28 also illustrates another embodiment of the mixing arrays of the invention. In this embodiment, used in conjunction with the gradient-forming embodiment described above, a specialized mixed fluid receiving chamber, termed a gradient chamber, is provided; one embodiment of this receiving chamber is shown in
Figure 28. This chamber permits the gradient fluid stream to be aliquotted into the individual compartments of the chamber, wherein the concentration of the gradient decreases with increasing distance from the gradient chamber inlet capillary. With gradients constituting decreasing concentrations of an analyte, drug, toxin or other species to be tested, the chamber can be modified to contain a detection means in each compartment, so that the concentration effect of the changing component of the gradient can be determined.
In yet another embodiment of the microfluidics platforms of this invention are provided a microsystems platform specifically designed for performing a specific binding assay. These embodiments are exemplified using an immunoassay as illustrated in Figures 25 through 27. In Figure 25, the arrangement of one assay array 19 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform. The components of the mixing array are shown in greater detail in Figure 26. It will be understood by a comparison of Figures 25 and 26 that the center of the platform 11 is at the top of Figure 26, and the edge or lateral extent of the platform is at the bottom of Figure 26, illustrated by a curved line. Rotation of the mixing array on platform disks of the invention can be in either direction, although rotation in a consistent, particular direction is preferred. Disk embodiments of the platforms of the invention were fashioned from machined acrylic. The overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device. The thickness of the disc ranged from about 0.9mm to about 1.5mm. The working fluid volume for reaction was about 10-100μL. In this aspect of the platforms of the invention there is provided an incubation chamber that comprises a specific binding reagent. For the purposes of this invention, the term "specific binding reagent" is intended to encompass biomolecules having a specific binding affinity between pairs thereof providing a specific molecular binding interaction with a binding affinity constant of between about 10~4 and 10 15M. Examples of such pairs of specific binding reagents include but are not limited to antigen and antibody, including antisera, polyclonal antibodies and most preferably monoclonal antibodies; receptor and ligands, including cell-surface receptors; integrins and adhesion proteins, including ICAM-I and ICAM-II; and carbohydrates and lectins, including phytohemagglutinin. As provided by the invention, specific binding reagents comprising a first member of a specific binding pair is provided in an incubation chamber as a coating on the surface of the chamber, as a dried or lyophilized reagent contained in the chamber and reconstituted upon addition of a fluid or fluid sample to the chamber; contained on a support media, such as latex or other beads, or in a gel or other support media. Said first member of a specific binding pair is designed or intended to detect the presence of an analyte, for example, a cell expressing a cognate antigen, receptor or adhesion protein or having a carbohydrate moiety at the cell surface specific for a particular lectin. Said specific binding reagent is applied to the incubation chamber of the platform by depositing the reagent on the surface using any appropriate means, including inkjet printing, computer-positioned syringes, microetching and microlithographic methods, including photolithography, screen and airbrush printing methods, solution coating, dipping, and conventional microtitre-well techniques. In applying said specific binding reagent, the surface or detection chamber can be treated to provide a two-dimensional array or pattern, wherein certain areas on the surface or detection chamber are treated with said specific binding reagent and others are not in a recognizable manner. The components of the specific binding assay array are as follows. An entry port
901 having a depth in the platform surface ranging from about 0.1mm to about lmm deep and lateral dimensions of about 0.1cm to about 2cm is constructed on the platform, and designed to accommodate a volume of about from 2μL to lOOμL. This entry port is fluidly connected with a metering capillary 902 having cross-sectional diameter of from about 0.1mm to about lmm, and having a depth of about 0.25mm to lmm; the length of this metering capillary was sufficient to contain a total volume of about 2 to about 1 OOμL. The metering capillary 902 is fluidly connected to capillary junction 904. The entry port is also constructed to be fluidly connected with an overflow capillary 903 having a cross-sectional diameter of about 0.1mm to about lmm and proximal ends rounded with respect to entry port 901. The overflow capillary is fluidly connected with an overflow chamber 905 having a depth in the platform surface of about
0.1mm to about lmm, greater than the depth of the overflow capillary 903. Each of the overflow and fluid chambers is also connected with air ports or air channels, such as 523, that have dimensions of about 0.1mm to about lmm and that permit venting of air displaced by fluid movement on the platform. Capillary junctions 524 that are about 0.75mm deep are present in the air channels to prevent fluid flow into the air channels.
Entry port 901 is positioned on the platform from 1cm to 20cm from the center of rotation. Metering capillary 902 extends from entry port 901 from about 0.5cm to about 5cm. Overflow capillary 902 extends from entry port 901 from about 1cm to about 20cm. The extent of the length of overflow capillary 903 is 20% longer than metering capillary 902. The position of overflow chamber 905 is from about 1cm to about 20cm from the center of rotation, and the position of capillary junction 904 is from about 1cm to about 20cm from the center of rotation.
Capillary junction 904 is fluidly connected with capillary channel 906, which in turn is fluidly connected with incubation chamber 910. Capillary channel 906 has a cross-sectional diameter of about 0.1mm to about lmm and extends from about 0.2cm to about 10cm from the capillary junction. Incubation chamber 910 has a depth in the platform surface ranging from about 0. lmm to about lmm, that is greater than the depth of capillary channel 906. Capillary channel 906 is also fluidly connected with channel 909 through capillary junction 907. Capillary junction 907 is constructed to prevent fluid flow backwards through the junction. Channel 909 has a cross-sectional diameter of about 0.1mm to about lmm and extends from about 0.2cm to about 5cm from the capillary junction. Capillary junction 907 has a depth in the platform surface of about 0.1 mm to about 1 mm, greater than the depth of the channel 909 or capillary channel 906. Incubation chamber 910 also contains a specific binding species, most preferably an antibody, specific for a component of the sample. This species is advantageously contained within incubation chamber 910 as a coating on the surface of the chamber, or attached to beads or other carrier within the chamber, or to a functionalized inner surface of the chamber or otherwise as described above.
Capillary junction 907 is further fluidly connected with wash buffer reservoir 516, having a depth in the platform surface of from about 0.1mm to about lmm and positioned at a distance from about 10mm to about 200mm from the axis of rotation.
Capillary junction 907 is further fluidly connected with reagent capillary 920, which is further fluidly connected with capillary junction 914, which is further fluidly connected with channel 926, and which is fluidly connected with reagent reservoir 917. Reagent capillary 920 has a cross-sectional diameter of about 0.1mm to about lmm and extends from about 0.2cm to about 20cm reagent reservoir 917. Capillary junction 914 has a depth in the platform surface ranging from about 0.1mm to about lmm and is positioned at a distance from about 10mm to about 200mm from the axis of rotation.
Reagent capillary 926 has a cross-sectional diameter of from about 0.1mm to about lmm and extends from about 0.2cm to about 20cm from the capillary junction. Reagent reservoir 917 has a depth in the platform surface of about 0.1mm to about lmm, and is positioned at a distance from about 10mm to about 200mm from the axis of rotation.
Incubation chamber 910 is fluidly connected at a point most distal to the axis of rotation to U-shaped capillary 921. U-shaped capillary 921 has a cross-sectional diameter of about 0.1mm to about lmm and extends from about 0.2cm to about 20cm between incubation chamber 910 and waste reservoir 915 . This capillary extends in a
U-shape to a point that is at least as proximal to the axis of rotation than the most axis- proximal extent of incubation chamber 910. This positioning of the U-shaped channel relative to incubation chamber 910 ensures that additional fluids flowing into incubation chamber 910 and displacing fluid therefrom will displace said fluid homogeneously, i. e. , the first fluid in the chamber will be pushed out of the chamber whilst being replaced by the second fluid.
This U-shaped capillary is also fluidly connected with waste reservoir 915. Waste reservoir 915 has a depth in the platform surface of about 0.1mm to about 5mm and is positioned at a distance from about 10mm to about 200mm from the axis of rotation; as shown in Figure 26, the waste reservoir is typically positioned at the farthest distance from the axis of rotation of any of the components of the array. In certain embodiments of the invention, sacrificial valves 922 can be positioned at the junction of capillary junction 904 and capillary channel 906, at the junction of capillary junction 907 and wash buffer capillary 908, or at the junction of reagent reservoir 918 and capillary junction 919. The use of this platform for performing an immunoassay is illustrated in Figures
27 A through 27L. In the use of this platform reagent reservoir 916 and wash reservoir 915 are pre-loaded on the disk, and most preferably the disk contains sacrificial valves 922 at the junction of capillary junction 907 and wash buffer capillary 908, and at the junction of reagent reservoir 918 and capillary junction 919. An imprecise volume (ranging from 1-15 OμL of fluid) of a fluid is applied to the entry port 901 (Figure 27A).
Fluid wicks into metering capillary 902 and stops at the capillary junction between metering capillary 902 and capillary junction 904 (Figures 27B and 27C). After sample loading by a user and filling of metering capillary 902 and overflow capillary 903 at no rotational speed, the platform is spun at a first rotational speed f,, ranging from 100- 500rpm; the exact value is dependent on the position of the components on the platform.
Due to the greater distance of the end of overflow capillary 903 from the center of rotation than the end of metering capillary 902, fluid flows through overflow capillary
903 into overflow chamber 905 (Figure 27D). The platform is spun until all excess fluid is evacuated from entry port 901 and into overflow chamber 905, except the fluid contained in metering capillary 902 (Figure 27E).
At a second rotational speed f2, that is greater than the first rotational speed f,, typically in the range of 100-1 OOOrpm, the capillary junction 904 at the distal end of the metering capillary 902 is overcome, and sample from metering capillary 902 fills incubation chamber 910 (Figures 27F and 27G). A portion of the sample wicks into U- shaped capillary 914 to the level of the sample in incubation chamber 910 (Figure 27G).
The sample is incubated for a time sufficient for maximum saturation binding of the component in the sample that specifically bind to the specific binding species.
At a third rotational speed f3, that is greater than the second rotational speed f2, typically in the range of 100-1500rpm, the capillary junction 908 is overcome, and wash buffer from reservoir 916 flows through capillary 909, capillary 906, and into incubation chamber 910. Wash buffer fluid flow forces the sample through U-shaped capillary 914 and into waste reservoir 915 (Figures 27 H through 27J). Preferably, sacrificial valves 922 are released to permit wash buffer fluid flow.
At a fourth rotational speed f4, that is greater than the third rotational speed f3, typically in the range of 100-2000rpm, the capillary junction 919 is overcome, and reagent buffer from reservoir 917 flows through capillary 918, capillary 920, capillary junction 908, capillary 906, and into incubation chamber 910. Reagent buffer fluid flow forces the wash buffer through U-shaped capillary 914 and into waste reservoir 515 (Figures 27 H through 27J). Preferably, sacrificial valves 922 are released to permit reagent buffer fluid flow. The reagent buffer contains a chromogen or other developing agent for detection of specific binding in incubation chamber 910.
2. Resistive Heater and Temperature Sensing Components Temperature control elements are provided to control the temperature of the platform. The invention provides heating elements, specifically resistive heating elements, and elements for detecting temperature at specific positions on the platform. Heating devices are preferably arrayed to control the temperature of the platform over a particular and defined area, and are provided having a steep temperature gradient with distance on the platform from the heater.
Certain resistors, including commercially- available resistive inks (available from Dupont) exhibit a positive temperature coefficient (PTC), i.e., an increase in resistance with increasing temperature. Applying a fixed voltage across a PTC resistor screen- printed on a plastic substrate results in rapid heating, followed by self-regulation at an elevated temperature defined by the circuit design heat sink and ambient temperature.
In such screen-printed resistors, connection to a power source is made by first printing parallel silver conductors followed by printing the PTC ink between the conductors as shown in Figure 29.
As shown in Figure 29, a resistive heating element comprises a conductive ink connected with electrical contacts for activation of the heater, and resistive inks applied between the conductive ink and in electrical contact therewith, wherein application of a voltage (direct or alternating cuπent) between the conductive inks results in current flow through the resistive inks and production of heat. There are two important types of resistive inks used in the resistive heating elements of this invention The first is a standard polymer thick film ink, such as Dupont 7082 or Dupont 7102 ink. These inks produce a surface temperature that is not self-limiting, and the temperature resulting from the use of these inks is dependent primarily on the magnitude of the applied voltage. In contrast, the positive temperature coefficient (PTC) inks show increase resistivity with increasing voltage, so that surface temperature is self-limiting because the amount of heat-producing current goes down as the applied voltage goes up. PTC inks are characterized as having a particular temperature where this self-limiting property is first exhibited; at voltages that produce temperatures less than the critical temperature, the amount of heat is dependent on the magnitude of the applied voltage.
Resistive inks useful according to the invention include Dupont 7082, 7102,
7271, 7278 and 7285, and other equivalent commercially available polymer thick film ink and PTC inks.
Conductive inks useful according to the invention include Dupont 5028, 5025, Acheson 423SS, 426SS and SS24890, and other equivalent commercially available conductive inks.
Additional components of the dielectric layer that serves to insulate the electrical circuit. Dielectric layers advantageously comprise dielectric inks such as Dupont
5018A. Insulation can also be achieved using pressure sensitive transfer adhesive such as 7952MP (3M Co.), or a pressure sensitive transfer adhesive deposited onto a polyester carrier layer such as 7953MP (3M Co.) or thermoplastic bonding films such as 3M 406, 560 or 615. Resistive heaters of the invention are advantageously used to incubate fluids at a stable temperature and for melting sacrificial valves as described below, and also for thermal cyclic.
Resistive and conductive inks are preferably screen-printed using methods and techniques well known in the art. See Gilleo, 1995, Polymer Thick Film (Van Nostrand Reinhold). Inks are typically screen printed to a thickness of about 10 microns; however, repetitive screen printing of resistive inks can be used to deposit thicker layers having reduced resistances. Both conductive and resistive inks are heat cured, typically at between 110 °C and 120 °C for about 10 minutes. The outline of this printing process is shown in Figure 30. Importantly, each of the layers must be correctly registered with one another for resistive heating to be provided. Heaters can be screen printed to any required size; a minimum area for a screen-printed heater has been determined to be about 0.25mm2 (0.5mm x 0.5mm).
The ability to tailor the resistance (and hence the temperature profile) of the resistive heaters using choice of ink formulation and reprinting of heater circuits provides control of the final electrical and thermal properties of the resistive heating elements of the invention. The resistance can also be controlled through connection of series and parallel configurations of resistive elements. For example, the particular circuits shown in Figure 31 allow for many parallel resistive elements per unit area; other configurations can be chosen for other applications.
3. Sacrificial Valves
The ability to specifically generate heat at a particular location on a microsystems platform of the invention also enables the use of sacrificial valves that can be released or dissolved using heat. For the purposes of this invention, the term "sacrificial valve" is intended to encompass materials comprising waxes, plastics, and other material that can form a solid or semi-solid fluid-tight obstruction in a microchannel, capillary, chamber, reservoir or other microfluidics component of the platforms of the invention, and that can be melted or deformed to remove the obstruction with the application of heat. Sacrificial valves are preferably made of a fungible material that can be removed from the fluid flow path. In preferred embodiments, said sacrificial valves are wax valves and are removed from the fluid flow path by heating, using any of a variety of heating means including infrared illumination and most preferably by activation of resistive heating elements on or embedded in the platform surface as described herein. For the purposes of this invention, the term "wax" is intended to encompass any solid, semi-solid or viscous liquid hydrocarbon, or a plastic. Examples include mondisperse hydrocarbons such as eicosane, tetracosane and octasone, and polydisperse hydrocarbons such as paraffin. In the use of wax sacrificial valves, application of a temperature higher than the melting temperature of the wax melts the valve and removes the occlusion from the microchannel, capillary or other fluidic component of the microsystems platforms of the invention. Particularly when the sacrificial valve is melted on a rotating microsystems platform of the invention, the melted wax to flow through the microchannel, capillary or other fluidic component of the microsystems platforms of the invention and away from the original site of the valve.
One drawback, however, is the possibility that the wax will recrystallize as it flows away from the original valve site, and concomitantly, away from the localized heat source. Recrystallization results in re-occlusion of the microchannel, capillary or other fluidic component of the microsystems platforms of the invention, potentially and most likely at a site other than the site of a localized heat source, and therefore likely to foul fluid movement on the disc. One solution for this problem is the inclusion in the sacrificial wax valves of the invention of a wax recrystallization chamber positioned "downstream" from the position of the wax valve. Preferably, the wax recrystallization chamber is fluidly connected with the microchannel, capillary or other fluidic component of the microsystems platforms of the invention that was occluded by the wax sacrificial valve. Typically, the wax recrystallization chamber is a widening of the microchannel, capillary or other fluidic component of the microsystems platforms of the invention so that recrystallized wax can harden on the walls of the microchannel, capillary or other fluidic component of the microsystems platforms of the invention with enough distance between said walls that the recrystallized wax does not re-occlude the microchannel, capillary or other fluidic component of the microsystems platforms of the invention. Preferably, the heating element, most preferably the resistive heating element of the invention, extends past the site of the wax valve and overlaps at least a portion of the wax recrystallization chamber, thereby retarding the propensity of the wax valve to recrystallize.
It is also recognized that this propensity of wax valves to recrystallize can be exploited to create a wax valve at a particular location in a microchannel, capillary or other fluidic component of the microsystems platforms of the invention. In this embodiment, a particular location can be kept below a threshold temperature by failing to apply heat at that location, and a wax valve material can be mobilized from a storage area on a platform by heating and them allowed to flow under centripetal acceleration to a particularly "cold" site where a wax valve is desired. An advantage of wax valves in this regard is that the proper positioning an activation of resistive heater elements enables flexibility in choosing when and whether a particular microchannel, capillary or other fluidic component of the microsystems platforms of the invention is to be occluded by a wax sacrificial valve.
In particularly preferred embodiments, the sacrificial valves of the invention comprise a cross-linked polymer that displays thermal recover, most preferably a cross- linked, prestressed, semicrystalline polymer; an example of a commercially available embodiment of such a polymer is heat recoverable tubing (#FP301H, 3M Co.,
Minneapolis, MN). Using these materials, at a temperature less than the "melting" temperature (Tm), the polymer occludes a microchannel, capillary or other fluidic component of the microsystems platforms of the invention. At a temperature greater than Tm„ however, the polymer reverts to its pre-stressed dimensions by shrinking. Such shrinking is accompanied by release of the occlusion from the microchannel, capillary or other fluidic component of the microsystems platforms of the invention. Such embodiments are particular preferred because the polymer remains in situ and does not recrystallize or otherwise re-occlude the microchannel, capillary or other fluidic component of the microsystems platforms of the invention. Also, such embodiments do not require the more extensive manipulation in preparing the platforms of the invention that wax valves require.
In another embodiment, the sacrificial valves of the invention comprise a thin polymeric layer or barrier dividing two liquid-containing microchannel, capillary or other fluidic component of the microsystems platforms of the invention, that can burst when sufficient temperature and/ or pressure is applied.
Another embodiment of the sacrificial valves of the invention are provided wherein a screen-printed resistive heater element is itself a valve. In this embodiment, the resistive heater element is screen-printed on a substrate such as polyester that divides two liquid-containing microchannel, capillary or other fluidic component of the microsystems platforms of the invention. In these embodiments, localized application of heat using a resistive heating element is used to melt the substrate dividing the liquid- containing microchannel, capillary or other fluidic component of the microsystems platforms of the invention. Preferably, in this embodiment the two liquid-containing microchannel, capillary or other fluidic component of the microsystems platforms of the invention are positioned in adjacent layers through the vertical thickness of the platform. As described above, the screen-printed resistive heater elements of this invention provide localized application of heat to a microsystems platform. The degree of localization achieved using these resistive heating elements is sufficient to provide for the placement of two adjacent sacrificial valves separated by a distance of 0.15cm.
4. Electrical Connection through a Slip Ring Rotor
The invention also provides a specialized rotor spindle for making electrical connection through a spindle to a rotating structure. An example of the specialized spindle structure of the invention is shown in Figure 1. The spindle is provided with a series of electrically conductive rings, each of which are electrically isolated from the others. Each conductive ring is led to contacts that impinge upon circuits resident on the rotating structure. Electrical power or data signals are delivered to and from the apparatus by means of conductive brushes that contact the conductive rings. Signals may be conducted while the apparatus is rotating or when stationary. Rotating structures such as centrifugal rotors and the microsystems platforms of this invention containing internal channels and reservoirs are used to control fluid movement. Such disks and rotors are known in the art as centrifugal analyzers and have been used to separation chemical and biochemical species as well as enabling the synthesis of chemical and biochemicals. However, a limitation of conventional centrifugal analyzers is that those analytical or synthetic procedures that required electrical input on the rotating structure itself were not enabled. Instead, mechanical and non-mechanical processes were used to regulate fluid movement in centrifuge rotors and other centripetally-motivated devices. Mechanical processes included valve actuation and pumping liquid toward the center of rotation; non-mechanical processes included heating, cooling, electrophoresis, and sensing through combinations of optical, electrical, chemical and biological means. The mechanical and non-mechanical processes known in the prior art were not capable of precisely regulating fluid movement when compared with equivalent, non-rotating devices that used electrical power.
Rotating structures permit precise control of fluid movement. Conventional centrifugal fluid control is restricted to outward flow in a radial direction that is primarily regulated through liquid placement (such as volume, positional radius, and liquid height), channel geometry (including considerations of channel radius) and rotational rate.
Temperature control is achieved over the entire rotor, by refrigerating the centrifugal compartment, for example. However, many chemical and biological processes require precisely controlled elevated temperature for optimal performance; for example, enzymatic reactions such as those used in immunoassays often have temperature optima of about 37 °C. In vitro amplification reactions (such as polymerase chain reaction) require even more elevated temperatures, such as cycling between 70 °C and 95°C.
A rotating structure presents unique difficulties in terms of thermal control, particularly heating. Conventional options included placement of the centrifuge rotor inside of an oven, subjecting one area to infrared radiation, and clamping the device between thermally controlled platens. Each of these approaches present difficulties. Ovens have unpredictable thermal gradients that prevent tight control of temperature 'in the interior of a rotating structure. Infrared presents similar difficulties, while clamping precludes spinning while heating unless the entire heating apparatus is also spun. All of these approaches suffer from an inability to directly measure the temperature inside the rotating structure. These reasons and other have prevented full integration of centripetally-motivated separation with analytical procedures in a single centrifugal rotor; typically, separations and analysis are performed separately in the prior art.
This invention provides a solution to this problem, specifically by enabling the transmission of electrical signals between a rotating structure such as a centrifuge rotor or microsystem platform of this invention and a fixed position electrical source. Electrical devices that can be advantageously controlled on a spinning rotor include temperature regulation devices, sensors, electrophoretic devices, integrated circuits and mechanical valves. These structures in turn permits a wider array of chemical processes to be performed on a disk than those permitted with current technology.
The invention provides a specialized rotor spindle for transmitting electrical signals between a rotating centrifuge rotor and a stationary electrical source as follows; the structure of the spindle will be best appreciated with reference to Figure 1. Into an electrically non-conductive plate is embedded a plurality of electrically conductive posts. The non-conductive plate is made of any insulating, electrically non-conductive material, including most preferably hardened plastics, rubber, WHAT ELSE??, and other electrical insulators and non-conductors. The electrically conductive posts are made from any electrically conducting metal, including copper, aluminum, WHAT ELSE???, and are preferably spring-loaded from the bottom of the non-conductive plate. A mechanical spindle is positioned on the side of the non-conductive plate opposite to the side of the plate containing the conductive posts. This plate is referred to as a spindle plate. A number of conductive disks equal to the number of conductive posts are layered on the spindle plate. The electrical contacts in the conductive plate are arranged so that each conducive plate is in electrical contact with a single conductive post. For the purposes of this invention, the term "in electrical contact" is intended to mean that an electric current can be produced through the electrical contacts at voltages (direct or alternative current) that may be effectively achieved in a centrifugal apparatus as described. Each conductive disk is similarly electrically isolated from other disks with interspersed sheets or disks of nonconducting material, preferably the same non- conductive material used to make the non-conductive plate. The last layer positioned farthest way from the spindle plate is a nonconductive layer and is arranged so that the tips of the conductive posts are at the surface or emerge from this plate. This final plate containing the ends of the conductive posts is referred to as the contact plate. An assembled unit, consisting of baseplate, posts, conductive element within the stack, is referred to as a signal channel. Typically one channel will be reserved as a common or ground channel.
The contact stack is held within a chassis that permits a series of electrically conductive brushes to impinge upon each of the conductive disks of the contact stack.
These brushes are arranged such that an electric signal can be propagated from one brush to one contact on the contact plate independently of electrical signals conducted from any other brush. The spindle is held in a spring-loaded bearing assembly such that the contact stack can rotate freely and present resistive pressure when pressed from below. The assembled unit is referred to as an electronic spindle. In the use of the electronic rotor of the invention, a centrifugal rotor or microsystems platform disk of this invention containing electrical contacts is aligned with the contacts on the contact plate of the electronic spindle in such a fashion that the electrical contact on the rotor are in electrical contact with the conductive posts on the contact plate of the electronic spindle. An electric signal can be propagated from the brushes of the electronic spindle to the contacts on the disk using the electronic spindle, most importantly when the rotor is spinning. Contact between the contact plate and the rotor is maintained by positive pressure provided by the spring-loaded bearing assembly and conductive posts. Electrical devices resident on the disk can then be monitored or controlled by signals through the electronic rotor. In a preferred embodiment, the rotor and the spindle contain complementary mechanical components that precisely align the rotor on the spindle to ensure proper electrical contact between the contacts on the rotor and the conductive posts on the spindle.
In a preferred embodiment thermal control structures resident on a rotating disk may be controlled through the electronic spindle. A resistive heating element prepared in a rotor as described herein is prepared having electrical contact leads positioned to be brought into electrical contact with one signal channel and the common channel of the electronic spindle. A thermistor element as described herein is placed on the disk in close proximity to the resistive heating element to enable thermistor response to be proportional to the temperature of the resistive heating element. The temperature of the heating element will be a function of the voltage and current applied to the disk through the electronic rotor and the speed at which the disk is being rotated (which will promote convective cooling). A thermal profile may be accurately monitored through the thermistor response and temperature controlled through adjustment of heating voltage and disk speed using the electronic spindle of the invention. In another preferred embodiment, the polymerase chain reaction ("PCR"') can be carried out on a rotating disk through thermal control using the electronic spindle. In this embodiment, a reaction chamber on a rotating disk is provided with the reagents necessary for PCR to provide a reaction mixture (see Saiki et al. , 1985 , Science : - ). This reaction chamber is in close proximity to a heater and thermistor as described herein to enable thermistor output to be proportional to the temperature of the reaction mixture. The reaction chamber is subjected to a cycling temperature profile sufficient to permit amplification of the template. Since the rate and precision at which a temperature change can be accomplished is a determining factor in the success of PCR amplification, control of voltage and speed of rotation as enabled using the electronic spindle of the invention permits PCR to be performed on a rotating platform.
In yet another preferred embodiment, other enzyme-requiring assays (such as enzyme linked immunosorbant assay ("ELISA")) can be performed and optimized on a rotating platform through thermal control using the electronic spindle of the invention. As those skilled in the art perform it, ELISA assays consist of antibody/antigen binding interactions followed by the conversion of a colorimetric or radioactive substrate into a detectable product. Detection is carried out through electrochemical or optical processes, depending the nature of the detectable product. Both antibody/antigen binding and enzymatic reactions have optimal temperatures (typically 37 °C) that can be achieved on a rotating platform using the resistive heater/ thermistor pair described above using the electronic spindle.
Electromechanical valves known in the prior art can also be incorporated in a rotating platform of centrifuge rotor using the electronic spindle of the invention. Mechanical, electrolytic or thermal valves on a rotating disk have been described (for example, in co-owned and co-pending U.S. Serial No. 08/761,063). By judicious activation of such valves the fractionation of complex mixtures can be carried out on a rotating platform. In preferred embodiments, complex biological mixtures such as milk or blood are subjected to centrifugal force to effect separation of constituent parts. Electromechanical, electrolytic or thermal valves can be opened, for example, to withdraw a fractionated supernatant away from a sedimented layer and into an adjacent chamber. Sample fractionation can be accomplished using repetitive sedimentation at different rotational rates, with appropriate valve activation used to partition portions of the fractionated to separate reservoirs or other compartments on the rotor. Upon fractionation the constituents may then be subjected to other processes such as PCR, immunoassay or electrophoresis.
Another application of the electronic spindle of the invention is to activate sensors on the rotating platform. In a preferred embodiment, sensors for detecting pH, are controlled and monitored through the electronic spindle.
The following Examples are intended to further illustrate certain preferred embodiments of the invention and are not limiting in nature.
EXAMPLE 1
Antibiotic Assay Disk
A microsystems platform provided by the invention and specifically designed for performing antibiotic assays is illustrated in Figures 1 and 2. Disk embodiments of the platforms of the invention were fashioned from machined acrylic and injection-molded polycarbonate. The overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk was mounted on the spindle of a rotary device. The thickness of the disc ranged from about 0.9mm to about 1.5mm.
The components of the antibiotic array were prepared as follows. An entry port 201 having a depth in the platform surface from about 0.75mm and lateral dimensions of from about 0.2cm to about 2cm were constructed on the platform, and designed to accommodate a volume of about 60μL. This entry port was fluidly connected with an array of eight metering capillaries 202 having a square cross-sectional diameter of about 0.5mm and proximal ends rounded with respect to entry port 201; the length of this metering capillary array was sufficient to contain a total volume of about 20μL. The entry port was also constructed to be fluidly connected with an overflow capillary 203 having a cross-sectional diameter of from about 0.02mm to about 0.75mm and proximal ends rounded with respect to entry port 201. The overflow capillary was fluidly connected with an overflow chamber 205 having a depth in the platform surface of about 0.75mm, greater than the depth of the overflow capillary 203. Metering capillary 202 was fluidly connected to fluid chamber 204 having a depth in the platform surface of about 0.63mm and greater than the depth of the metering capillary 202. Each of the overflow and fluid chambers was also connected with air ports or air channels, such as 211, that have dimensions of about 0.25mm deep and permitted venting of air displaced by fluid movement on the platform. A capillary junction 212 that is about 0.75mm deep is present in the air channel to prevent fluid flow into the air channel. Entry port 201 was positioned on the platform from about 2cm from the center of rotation. Metering capillary 202 extended about 1 cm from entry port 201. The extent of the length of overflow capillary 203 was at least about 20%o greater than the extent of the length of metering capillary 202. The position of fluid chamber 204 was from about 3.2cm from the center of rotation, and the position of overflow chamber 205 was thus from about 5 cm from the axis of rotation.
The fluid chamber 204 acted as a capillary barrier that prevents fluid flow from metering capillary 202 at a first, non-zero rotational speed f, sufficient to permit fluid flow comprising overflow from the entry port 201 through overflow capillary 203 and into overflow chamber 205. This capillary boundary was constructed to be overcome at a second rotational speed f2 (where f2 > fi). Fluid chamber 204 was fluidly connected to capillary 206 that was 0.25mm deep and had a cross-sectional diameter of about 0.5mm and was connected to holding chamber 207. Holding chamber 207 had a depth in the platform surface of 0.75mm, greater than the depth of capillary 206. Filling of fluid chamber 204 is accompanied by fluid flow through capillary 206 into holding chamber 207. Holding chamber 207 was fluidly connected by way of capillary 208, having a square cross-sectional diameter of about 0.25mm and was connected with read chamber 210, having a depth in the platform surface of about 0.75mm, greater than the depth of capillary 208. In certain embodiments, a sacrificial valve 213 was placed as shown in the channel 209. As illustrated in Figures 3 A through 3J, in the use of this platform an imprecise volume (ranging from 20-60μL of fluid) of a fluid was applied to the entry port 201 (Figure 3 A). In embodiments of the platform comprising air displacement channels, the fluid wicked into air channel 211 and was stopped by capillary junction 212. Fluid also wicked into metering capillary 202 and overflow capillary 203. Fluid flowed through the metering capillary 202 and overflow capillary 203 at no rotational speed until the fluid reached capillary junctions at the junction between metering capillary 202 and fluid chamber 204 and overflow capillary 203 and overflow chamber 205 (Figures 3B and 3C). Metering capillary 202 was constructed to define a precise volume from about 20- 60μL of fluid between entry port 201 and the capillary junction at fluid chamber 204, which was designed to be at least the amount of the fluid placed by the user in entry port 201.
After sample loading by a user and filling of metering capillary 202 and overflow capillary 203 at no rotational speed, the platform was spun at a first rotational speed f]5 ranging from of 175rpm, which was sufficient for this microfluidics array having an entry port 201 with a depth of 0.6mm, metering capillary 202 with dimensions of 0.5mm x 0.5mm in cross-section and a length of 2.2-3.8cm from the center of rotation and an overflow capillary 203 with dimensions of 0.5mm x 0.5mm in cross-section and a length of 5.4cm from the center of.
Due to the greater distance of the end of overflow capillary 203 from the center of rotation than the end of metering capillary 202, fluid flowed through overflow capillary 203 into overflow chamber 205. The platform was spun until all excess fluid is evacuated from entry port 201 and into overflow chamber 205, except the fluid contained in metering capillary 202 (Figure 3D).
At a second rotational speed f2 of 360rpm, the precise amount of fluid contained in metering capillary 202 was delivered into fluid chamber 204 (Figures 3E through 3H) Fluid movement into fluid chamber 204 was accompanied by filling of capillary 206 and holding chamber 207.
In embodiments comprising a sacrificial valve 213 in-line with capillary 208 at position 209 shown in Figure 2, release of the sacrificial valve resulted in fluid flow into read chamber 210. In said embodiments, fluid flow is achieved at rotational speed f2 with removal of the sacrificial valve. In embodiments of the platforms of the invention comprising antibiotic arrays as described herein and not containing a sacrificial valve at position 209, capillary 208 preferably filled along with filling of holding chamber 207 until the fluid reached capillary junction 209 at the junction between capillary 208 and read chamber 210; in such embodiments, the capillary junction had a depth of about 0.75mm. At a third rotational speed f3 of about 520rpm, the fluid contained in holding chamber 207 was delivered into read chamber 210 (Figures 31 and 3J).
This embodiment of the microfluidics platforms of the invention was designed to use the carboxypeptidase inhibition assay described above for detecting beta-lactam antibiotics. The extent of chromogen production was detected in the read chamber, and related to the presence of antibiotics in the sample by comparison with samples tested in the absence of antibiotic. The amount of antibiotic in a test sample was determined using this platform of the invention.
EXAMPLE 2 Two-Step Assay Disk: Alternative Embodiment
In an alternative embodiment, a two-step assay disk of the invention was provided as shown in Figures 4 and 5. Disk embodiments of the platforms of the invention were fashioned from machined acrylic. The overall disc dimensions included an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk was mounted on the spindle of a rotary device. The thickness of the disc ranged from about
0.9mm to about 1.5mm. The working fluid volume for reaction with reagents was about 20μL.
The components of this two-step assay array were prepared as follows. An entry port 301 having a depth in the platform surface of about 0.75mm and lateral dimensions of from about 0.2cm to about 2cm was constructed on the platform, and designed to accommodate a volume of about 60μL. This entry port was fluidly connected with a multiplicity of entry capillaries 302 having a square cross-sectional diameter of about 0.5mm, having a depth of 0.5mm and proximal ends rounded with respect to entry port 301; the length of this entry capillary array was sufficient to contain a total volume of about 20μL. The entry capillaries 302 were fluidly connected to fluid chamber 303 having a depth in the platform surface of about 0.6mm, wherein the depth was greater than the depth of the entry capillary 302. Each of the fluid chambers of this aspect of the invention was also connected with air ports or air channels, such as 311, that have dimensions of about 0.25mm deep and permitted venting of air displaced by fluid movement on the platform. A capillary junction 312 that was about 0.75mm deep was present in the air channel to prevent fluid flow into the air channel. The fluid chamber 303 was also constructed to be fluidly connected with an overflow capillary 304 having a cross-sectional diameter of from about 0.02mm to about
0.75mm and proximal ends rounded with respect to fluid chamber 304. The overflow capillary was fluidly connected with an overflow chamber 306 having a depth in the platform surface of about 0.75mm, greater than the depth of the overflow capillary 304.
Entry port 301 was positioned on the platform about 1cm from the center of rotation. Entry capillaries 302 extends about 2cm from entry port 301. The position of a first fluid chamber 303 was from about 3cm from the center of rotation. The first fluid chamber 303 acted as a capillary barrier that prevented fluid flow from entry capillary 302 at zero rotational speed. Movement of fluid from entry port 301 through entry capillaries 302 and into the first fluid chamber 303 was achieved by rotation at a first, non-zero rotational speed f,. Displacement of fluid into the first fluid chamber 303 was accompanied by fluid filling of channel 305 that was fluidly connected with the first fluid chamber 303 and was positioned at the most radially distal point of the first fluid chamber. Channel 305 was fluidly connected with a second fluid chamber 307 and formed a capillary boundary between channel 305 and chamber 307. This capillary boundary was constructed to be overcome at second rotational speed f2 (where f2 > fj). First fluid chamber 303 was also fluidly connected to overflow capillary 304 that was 0.25mm deep and had a cross-sectional diameter of about 0.5mm and that extended from about 1cm to about 5cm and was connected to overflow chamber 306. Overflow chamber 306 had a depth in the platform surface equal to that of overflow capillary 304, so that there was no capillary boundary between overflow capillary 304 and overflow chamber 306. Overflow capillary 304 was positioned in the first fluid chamber 303 at a point radially less distant from entry port 301 than channel 305, thereby defining a volume in the fluid chamber between the position of the overflow capillary 304 and the most radially distant extent of the said first fluid chamber.
Second fluid chamber 307 was further fluidly connected through channel 308 to a small pocket or capillary junction 309. Channel 308, having a cross-sectional diameter of about 0.25mm and that extended from about 0.2cm to about 20cm, was fluidly connected to a third fluid chamber 310, having a depth in the platform surface of about 0.75mm, that was greater than the depth of capillary 308. Air recirculation channels 311 that had dimensions of about 0.25mm deep provided pathways for air displaced by fluid movement, while capillary junctions 312 that were about 0.75mm deep prevent fluid from entering the air channels. In some embodiments of the device a sacrificial valve 313 was placed as shown in Figure 5 in the channel 309. In certain embodiments, a valve 314 was placed in channel 305 to control fluid movement from the first fluid chamber 303 to the second fluid chamber 307.
As illustrated in Figures 6 A through 6J, in the use of this platform an imprecise volume (ranging from l-150μL of fluid) of a fluid was applied to the entry port 301 (Figure 6A). Fluid wicked into entry capillary 302 and stopped at the capillary junction between entry capillary 302 and the first fluid chamber 303 (Figures 6B and 6C). Fluid flowed through the entry capillary B and into the first fluid chamber 303 at a first rotational speed f, of 40rpm (Figures 6D and 6E). The fluid further entered capillary channel 305, stopping at the capillary junction with the second fluid chamber 307. As rotation continued, the fluid continued to fill the first fluid chamber 303, overflow capillary 304 fills (Figure 6F), and excess fluid filled overflow chamber 306 until the level of fluid in the first fluid chamber 303 fell below the position of overflow capillary 304 (Figure 6G).
At a second rotational speed f2 of 280rpm, the capillary junction between channel 305 and the second fluid chamber 307 was overcome, and fluid remaining in the first fluid chamber 303wasdelivered into the second fluid chamber 307 (Figures 6H and 61).
In an alternative embodiment, a sacrificial valve 314 was placed at the junction of channel 305 and the second fluid chamber 308, which sacrificial valve was released to permit fluid flow through channel 305 and into the second fluid chamber 308. In such embodiments, fluid flow can be achieved at either f, or f2 rotational velocity.
In embodiments comprising a sacrificial valve 313 in-line with capillary 308 at position 309 shown in Figure 5, release of the sacrificial valve results in fluid flow into the third fluid chamber 310. In said embodiments, fluid flow was achieved at rotational speed f2 with removal of the sacrificial valve. In embodiments of the platforms of the invention comprising two-step assay arrays as described herein and not containing a sacrificial valve at position 310, capillary 309 filled along with filling of the second chamber 308 until the fluid reached capillary junction 309 at the junction between capillary 308 and the third fluid chamber 310; in such embodiments, the capillary junction has a depth of about 0.75mm. At a third rotational speed f3 of about rpm, the fluid contained in the second chamber 308 was delivered into the third fluid chamber 310 (Figures 6H through 6K).
EXAMPLE 3 Blood Separation Array
A microsystems platform provided by the invention and specifically designed for separating vertebrate blood cells and components is illustrated in Figures 7 through 9.
The components of the blood separation array are shown in greater detail in
Figure 8. It will be understood by a comparison of Figures 7 and 8 that the center of the platform 11 is at the top of Figure 8, and the edge or lateral extent of the platform is at the bottom of Figure 8, illustrated by a curved line. Rotation of the blood separation array on platform disks of the invention can be in either direction, although rotation in a consistent, particular direction is preferred. Disk embodiments of the platforms of the invention were fashioned from machined acrylic. The overall disc dimensions included an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device. The thickness of the disc ranged from about 0.9mm to about 1.5mm. The working fluid volume for reaction with reagents was about
15μL.
The components of the blood separation array were as follows. An entry port 401 having a depth in the platform surface of about 0.5mm and lateral dimensions of about 0.5cm was constructed on the platform, and designed to accommodate a volume of about 20μL. This entry port was fluidly connected to an entry capillary 402, having a cross-sectional diameter of about 0.5mm and having a depth of 0.5mm; the length of this entry capillary was sufficient to contain a total volume of about 20μL. Entry capillary 402 was further fluidly connected to a separation column 403 having a cross- sectional diameter of about 1.25mm, a depth of about 0.75mm, and a length of this separation column was sufficient to contain a total volume of about 15μL. This separation column was also fluidly connected with a passage 411 to overflow chamber 404. Passage 411 had a cross-sectional diameter of about lmm, a depth of from about 0.5mm, and a length of 2mm. Overflow chamber 404 has a depth of about 0.5mm.
A small capillary exit 406 was also fluidly connected with separation chamber 403, having a cross-sectional diameter of about 0.125mm, a depth of about 0.125mm, and a length of about 0.75mm. This capillary was arranged to traverse a direction radially more proximal to the axis of rotation than the insertion point with separation column 403. This small capillary 406 terminated in a capillary junction 407 that was fluidly connected with capillary 408, extending in a radial direction to decant chamber
405. A sacrificial valve 413 is positioned in capillary 406 at the juncture with capillary junction 407. Capillary 408 had a cross-sectional diameter of about 0.25mm, a depth of about 0.25mm, and a length of about 3.5mm. This capillary was arranged in a radially outward direction between capillary junction 407 and decant chamber 405. Passage 411 was positioned on separation column 403 to be significantly more proximal to the axis of rotation than the insertion point of small capillary 406. Air displacement channels 409 that have dimensions of about 0.25mm deep permitted venting of air displaced by fluid movement on the platform. Capillary junctions 410 that were about 0.75mm deep were present in the air channels to prevent fluid flow into the air channels.
As illustrated in Figures 9A through 9H, in the use of this platform an imprecise volume (about 25μL) of blood was applied to the entry port 401 (Figure 9A). Blood entered the entry capillary 402 by capillary action, and stopped at the capillary junction between entry capillary 402 and the separation chamber 403 (Figures 9B and 9C).
At a first rotational speed f, of 150rpm, blood flowed from the entry capillary 402 into separation chamber 403 (Figure 9D). Blood continued to fill separation column 403 until blood reached the position of passage 411, whereupon excess blood flowed through passage 411 and into overflow chamber 404 (Figures 4E and 4F). Advantageously, small channel 406 had dimensions that prevent wicking of blood into the channel as blood flows past the insertion point of small channel 406 into separation column 403. As shown in Figure 9F, after sufficient time of rotation at the first non-zero rotational speed f„ the excess blood has been transferred into overflow chamber 404 and the separation column 403 was filled with blood to the position of passage 411. Rotation at a second rotational speed f2 of 1300rpm, blood components were separated into red blood cell, white blood cell (i.e., "buffy coat"), and plasma fractions (Figure 9G). Advantageous dimensions of small capillary 406 permitted fluid flow of the plasma fraction through capillary 406 that was stopped at capillary junction 407. Fluid flow of plasma into decant chamber 405 resulted from fluid flow overcoming the capillary barrier 407 by rotation at a third rotational speed f3 of about 1420rpm (Figure 9H).
EXAMPLE 4
Blood Separation Array: Alternative Embodiment
In an alternative embodiment, a blood separation disk of the invention is provided as shown in Figures 10 and 11. It will be understood that, as in Example 1, in Figure 10, the arrangement of one separation array 15 on a disk 11 is shown; a multiplicity of such arrays can be advantageously arranged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform. Disk embodiments of the platforms of the invention are fashioned from machined acrylic. The overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device. The thickness of the disc ranged from about 0.9mm to about 1.5mm. The working fluid volume for reaction with reagents is about 25μL.
The components of this separation array are as follows. An entry port 501 having a depth in the platform surface from about 0.75mm and lateral dimensions of from about 0.5cm is constructed on the platform, and designed to accommodate a volume of about 20μL. This entry port is fluidly connected with a first array of metering capillaries 502 and a second array of metering capillaries 503, wherein each of the capillaries has a cross-sectional diameter of about 0.5mm. The length of the second metering capillary array 503 is longer than that of the first metering capillary array 502. The first metering capillary array 502 is fluidly connected with a ballast chamber 507, having a depth in the platform surface of about 0.75mm and greater than the depth of the first metering capillary array 502, wherein the first metering capillary array 502 forms a capillary junction between the array and the ballast chamber. The second capillary aπay 503 is fluidly connected with capillary junction 506.
The entry port is also constructed to be fluidly connected with an overflow capillary 504 having a cross-sectional diameter of about 0.5mm and proximal ends rounded with respect to entry port 501. The overflow capillary is fluidly connected with an overflow chamber 505 having a depth in the platform surface of about 0.75mm, greater than the depth of the overflow capillary 504. Each of the overflow and fluid chambers is also connected with air ports or air channels, such as 514, that have dimensions of about 0.25mm deep and permit venting of air displaced by fluid movement on the platform. Capillary junctions 516 that is about 0.75mm deep are present in the air channels to prevent fluid flow into the air channels.
Entry port 501 is positioned on the platform from 1cm to 20cm from the center of rotation. Metering capillary array 502 extends from entry port 501 from about 0.6cm. Metering capillary array 503 extends from entry port 501 from about 1.9cm. The extent of the length of metering capillary array 503 is about 20%> longer than metering capillary array 502, and the extent of the length of overflow capillary 504 is at least about 20% greater than the extent of the length of either the first metering capillary array 502 or the second metering capillary array 503. The position of ballast chamber 507 is about 2.8cm from the center of rotation, the position of capillary junction 506 is about 3.8cm from the center of rotation, and the position of overflow chamber 505 is thus about 5cm from the axis of rotation.
The ballast chamber 507 acts as a capillary barrier that prevents fluid flow from the first metering capillary array 502 at a first, non-zero rotational speed fj sufficient to permit fluid flow comprising excess blood overflow from the entry port 501 through overflow capillary 504 and into overflow chamber 505. Capillary junction 506 is a capillary barrier that prevents fluid flow from the second metering capillary array 503 at said first, non-zero rotational speed f, sufficient to permit fluid flow comprising excess blood overflow from the entry port 501 through overflow capillary 504 and into overflow chamber 505. These capillary boundaries are constructed to be overcome at a second rotational speed f2 (where f2 > fj).
Ballast chamber 507 is fluidly connected to capillary 510 that is 0.25mm deep and has a cross-sectional diameter of about 0.65mm and that extends about 5cm and is connected to capillary junction 511. A sacrificial valve 518 is positioned at the exit of ballast chamber 507 at the juncture with capillary junction 511. Alternatively, capillary 510 is fluidly connected with a sacrificial valve 515. In said embodiments, fluid flow is achieved at rotational speed f2 with removal of the sacrificial valve. Sacrificial valve
515 or capillary junction 511 are further fluidly connected with channel 512 which is about 0.25mm deep and has a cross-sectional diameter of about 0.25mm and that extends about 3cm. Channel 512 is fluidly connected with separation chamber 509 at a point most distal from the axis of rotation. Second metering capillary array 503 is fluidly connected with capillary junction
506, which is overcome at a rotational speed f2 > f,. Capillary junction 506 is further fluidly connected to channel 508, which is further fluidly connected to separation chamber 509. Channel 508 is about 0.25mm deep and has a cross-sectional diameter of about 0.25mm. Separation chamber 509 is about 0.75mm deep and has a cross-sectional diameter of about 5mm, and is positioned from about 40cm from the center of rotation.
Separation chamber 509 is fluidly connected with decant channel 517 at a point close to the chamber's most axis-proximal extent. Decant channel 517 is about 0.25mm deep and has a cross-sectional diameter of about 0.25mm and extends about 4.3cm to about 5cm. Decant channel 517 is fluidly connected with decant chamber 514, which is about 0.75mm deep and has a cross-sectional diameter of about 5mm, and is positioned from about 50cm to about 80cm from the center of rotation..
As illustrated in Figures 12A through 12J, in the use of this platform an imprecise volume (about 25μL of fluid) of blood is applied to the entry port 501 (Figure 12 A). Blood enters the each of the metering capillary arrays 502 and 503 and stops at the capillary junction between metering capillary array 502 and ballast chamber 507 and between metering capillary 503 and capillary junction 506 (Figures 12B and 12C). Blood also enters and fills overflow capillary 504, stopping at the capillary junction with overflow chamber 505.
At a first rotational speed f, of 45rpm, blood flows from the entry port 501 through overflow capillary 504 and into overflow chamber 505 (Figures 12D and 12E).
At a second rotational speed f2 of 70rpm, the capillary junction between the first metering capillary array 502 and ballast chamber 508 is overcome, and blood from the first metering capillary array fills ballast chamber 508 (Figure 12F). Similarly, at second rotational speed f2, capillary junction 506 is overcome, and blood from second metering capillary array 503 enters separation chamber 509 (Figure 12F). Advantageously, the volume of blood in second metering capillary aπay 503 is insufficient to fill separation chamber 509 to the level of insertion of decant channel 517.
By rotation at a third rotational speed f3 of 1300rpm, blood components in separation chamber 509 are separated into red blood cell, white blood cell (i.e., "buffy coat"), and plasma fractions (Figures 12G and 12H). Separation of blood components is not achieved in ballast chamber 507, due to its position on the platform, and the capillary junction 51 lor sacrificial valve 515 are not overcome at third rotational speed f3. Advantageously, the separated plasma does not extend to decant capillary 517.
Release of sacrificial valve 517, or rotation at a fourth rotational speed f4 of rpm, results in flow of blood from ballast chamber 507 through channel 512 and into separation chamber 509 at the "bottom" or most axis-distal extent of the separation chamber (Figure 121). This results in filling of the separation chamber to a position equal to the insertion point of decant channel 517 (Figure 12J). Plasma flow through decant channel 517 and into decant chamber 514 in an amount equal to the amount of blood contained in ballast chamber 507. Decant channel 517 is advantageously provided with dimensions that retard passage of unfractionated blood, or plasma contaminated with greater than 0.1-1% of blood cells in whole blood.
EXAMPLE 5 Mixing Arrays A microsystems platform provided by the invention and specifically designed for performing mixing of equal volumes of different liquid samples is illustrated in Figure 13. In the Figure, the arrangement of one assay aπay 15 on a disk 11 is shown; a multiplicity of such aπays can be advantageously aπanged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform. The components of the mixing array are shown in greater detail in Figure 14.
Disk embodiments of the platforms of the invention were fashioned from machined acrylic. The overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device. The thickness of the disc ranged from about 0.9mm to about 1.5mm. The working fluid volume was about 50μL. The components of the mixing array were prepared as follows. Entry ports 601 having a depth in the platform surface of about 0.5mm and lateral dimensions of about lcm to about 5cm were constructed on the platform, and designed to accommodate a volume of about 5-50μL. Each entry port was fluidly connected with one of a paired aπay of metering capillaries 602 having a square cross-sectional diameter of about 0.5mm and proximal ends rounded with respect to entry port 601; the length of each metering capillary aπay was sufficient to contain a total volume of about 25μL. Metering capillaries 602 were fluidly connected to a curved capillary baπier 603 having a depth in the platform surface of about 0.25mm that was greater than the depth of metering capillaries 602. The capillary baπier 603 and other fluid components of the mixing aπay were also connected with air channels 608, that have dimensions of about
0.25mm deep and permitted venting of air displaced by fluid movement on the platform. In addition, capillary junctions 609 that were about 0.75mm deep were present in the air channels to prevent fluid backflow into the air channel.
Capillary baπier 603 was fluidly connected by a naπow capillary channel 604 to mixing chamber 605, which was fluidly connected with channel 610, which was further connected with mixed fluid receiving chamber 606. Alternatively, capillary 604 comprises a sacrificial valve 612. Capillary channel 604 was about 0.25mm deep and had a cross-sectional diameter of about 0.25mm and that extends about 0.2cm. Mixing chamber 605 was about 0.75mm deep and had a cross-sectional diameter of about 2mm, and was positioned about 4cm from the center of rotation. Capillary channel 610 was about 0.25mm deep and had a cross-sectional diameter of about 0.25mm and that extended from about 0.2cm to about 30cm. Mixing chamber 605 was constructed such that the insertion point of capillary channel 604 and the insertion point of capillary channel 610 were offset at opposite ends of the mixing chamber. As a consequence, fluid flowing through capillary channel 604 was forced to encounter the opposite wall of mixing chamber 605 before fluid flow can proceed through capillary channel 610. This results in the creation of turbulence in the mixed laminar fluid stream in capillary channel 604 caused by the conjoint flow of fluid from the first and second metering channels without appreciable mixing. The turbulence created by the structure of mixing chamber 605 was sufficient to disrupt laminar flow and cause fluid mixing in the chamber prior to continued fluid flow through capillary channel 610 and into mixed fluid receiving chamber 606. Mixed fluid receiving chamber 606 was about 0.75mm deep and had a cross-sectional diameter of about 5mm, and was positioned from about lcm to about 30cm from the center of rotation.
As illustrated in Figures 15 A through 15d, in the use of this platform an equal volume (ranging from l-150μL of fluid) of each of the fluids to be mixed was applied to the entry ports 601 (Figure 15 A). Fluid enters the each of the metering capillary aπays 602 and stops at capillary baπier 603.
At a first rotational speed f, of 90rpm, the fluids from each capillary aπay flowed into and filled the capillary baπier 603 (Figure 15B). In embodiments comprising a sacrificial valve 612, the valve prevented fluid flow into channel 604. Upon release of sacrificial valve 612 or rotation at first rotational speed f,, fluid flow proceeded from capillary junction 603 through channel 604 and into mixing chamber 605 (Figure 15C). Fluid flow within mixing chamber 605 was turbulent, in contrast to fluid flow through capillary baπier 603 or channel 604, which was primarily laminar, so that mixing occurs predominantly in mixing chamber 605. Fluid flow proceed through channel 610 and the mixed fluid solution was displaced into mixed fluid receiving chamber 606 (Figure 15D).
EXAMPLE 6 Mixing Arrays: First Alternatives
An additional embodiment of the microsystems platform provided by the invention and specifically designed for performing mixing of equal volumes (Figures 17 through 18) or unequal volumes (Figures 19 through 21) of different liquid samples. In these Figures, the aπangement of one assay aπay 17 on a disk 11 is shown; a multiplicity of such aπays can be advantageously aπanged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform. The components of the mixing aπay are shown in greater detail in Figures 17 and 20. Disk embodiments of the platforms of the invention are fashioned from machined acrylic. The overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device. The thickness of the disc ranged from about 0.9mm to about 1.5mm. The working fluid volume is about 40μL in each fluid reservoir.
The components of the mixing aπay are as follows. Mixing of equal volumes is illustrated in Figure 17 and for unequal volumes in Figure 20. (Components will be identified using the number from Figure 17; equivalent structures are shown in Figure 20 for unequal mixing and will be shown in parentheses). Fluid reservoirs 651 and 652 (701 and 702), each containing one of a pair of liquids to be mixed, are constructed on the platform, having a depth in the platform surface from about 0.75mm and lateral dimensions of lcm, and wherein fluid reservoirs 651 and 652 are designed to accommodate equal volumes of fluid (about 50μL); (for unequal volumesn fluid reservoir 701 is designed to accommodate a volume of about 45 μL, and fluid reservoir 702 is designed to accommodate a volume of about 5μL, wherein the volume of fluid reservoir 702 is less than the volume of fluid reservoir 701.) In particular and in addition, the viscosity of the fluid in the fluid reservoirs may differ, so that mixing produces a mixed fluid of intermediate viscosity. Each fluid reservoir is fluidly connected with a capillary channel 653 and 654 (703 or 704) to capillary junction 655 (705). Each capillary channel is about 0.5mm deep and having a cross-sectional diameter of about 0.5mm and that extends about 5cm. Capillary junction junction 655 (705) has a depth in the platform surface of about 0.75mm that is greater than the depth of capillaries 652 and 653 (703 and 704). Alternatively, capillaries 652 and 653 (703 and 704) comprise a sacrificial valve 662 (712). Use of said sacrificial valves can be used in addition to or in place of capillary junction 655 (705).
The fluid components of the mixing aπay are also connected with air channels 660 (710), that have dimensions of about 0.25mm deep and permit venting of air displaced by fluid movement on the platform. In addition, capillary junctions 661 (711) that are about 0.75mm deep are present in the air channels to prevent fluid backflow into the air channel.
Capillary junction 655 (705) is fluidly connected by a naπow capillary channel 656 (706) to mixing chamber 657 (707), which is fluidly connected with channel 658 (708) which is further connected with mixed fluid receiving chamber 659 (709). Alternatively, capillary 656 (706) comprises a sacrificial valve 662 (712). Capillary channel 656 (706) is about 0.25mm deep and having a cross-sectional diameter of from about 0.5mm and that extends from about 0.2cm to about 30cm. Mixing chamber 657 (707) is about 0.25mm deep and having a cross-sectional diameter of from about 0.75mm, and is positioned from about 0.2cm to about 30cm from the center of rotation. Capillary channel 658 (708) is about 0.5mm deep and has a cross-sectional diameter of about 5mm and that extends from about 0.2cm to about 30cm. Capillary channel 656 (706) and capillary channel 658 (708) may be offset in their connection with the mixing chamber as previously described in Example 5 , or the capillaries can be positioned in the mixing chamber at any convenient position, and Coriolis forces have be relied upon to facilitate mixing.
Capillary 658 (708) is fluidly connected with mixed fluid receiving chamber 659 (709). Mixed fluid receiving chamber 659 (709) is about 0.75mm deep and having a cross-sectional diameter of about 5mm from about, and is positioned from about lcm to about 30cm from the center of rotation.
As illustrated in Figures 18E through 18E for mixing equal volumes, and in Figures 21 A through 21E for mixing unequal volumes, in the use of this platform a volume of each of the fluids to be mixed is applied to the fluid reservoirs 651 and 652
(701 and 702) (Figures 18A and 21 A). Fluid enters the each of the capillaries 653 and 654 (703 or 704) and stops at capillary junction 655 (705). Alternatively, the platforms of the invention are provided containing the fluids to be mixed already in fluid reservoirs 651 and 652 (701 and 702). In these embodiments, it is prefeπed that sacrificial valves 712 be provided in capillaries 653 and 654 (703 or 704), to prevent evaporation, wetting or leakage of fluid from the reservoirs prior to use.
At a first rotational speed f, of lOOrpm, the fluids from each capillary flows past capillary junction 655 (705) and through mixing chamber 657 (707) (Figures 18B and 18C and Figures 21B and 21C). In embodiments comprising a sacrificial valve 712, the valve prevents fluid flow into channels 653 and 654 (703 or 704). Upon release of sacrificial valve 712, fluid flow proceeds from capillary junction 655 (705) through channel 656 (706) and into mixing chamber 707 (Figure s 18C and 21C). Fluid flow within mixing chamber 657 (707) is turbulent, in contrast to fluid flow through capillary barrier 655 (705) or channel 656 (706), which is primarily laminar, so that mixing occurs predominantly in mixing chamber 657 (707). Fluid flow proceed through channel 658 (708) and the mixed fluid solution is displaced into mixed fluid receiving chamber 659 (709) (Figures 18D and 18 E and Figures 21D and 21E). EXAMPLE 7 Mixing Arrays: Second Alternative
An additional embodiment of the microsystems platform provided by the invention and specifically designed for performing mixing of different volumes of liquid samples to form a gradient in the concentration of a species in which the two fluids differ; this embodiment is illustrated in Figure 22. In the Figure, the aπangement of one assay aπay 18 on a disk 11 is shown; a multiplicity of such aπays can be advantageously aπanged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform. The components of the mixing aπay are shown in greater detail in Figure 23.
Disk embodiments of the platforms of the invention are fashioned from machined acrylic. The overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device. The thickness of the disc ranged from about 0.9mm to about 1.5mm. The working fluid volume is about 40μL for each fluid reservoir.
The components of the mixing aπay are as follows. Fluid reservoirs 801 and 802, each containing one of a pair of liquids to be mixed, are constructed on the platform, having a depth in the platform surface from about 0.75mm and lateral dimensions of lcm, and wherein fluid reservoir 801 is designed to accommodate a volume of about 40μL, and fluid reservoir 802 is designed to accommodate a volume of about 40μL, wherein the volume of fluid reservoir 802 is different than the volume of fluid reservoir 801. In particular and in addition, fluid reservoirs 801 and 802 are shaped so that the rate of fluid output in the two reservoirs differs between the reservoirs at a particular rotational speed, due to a change in the pressure "head" (related to the cross-sectional area of the fluid at each point in the reservoir), so that the proportion of fluid in the mixture from one of the reservoirs is at a maximum at the beginning of rotation and is at a minimum when the fluids from the reservoirs are completely mixed at the end of rotation, thus forming a gradient. Gradient should be approximately 40% of left reservoir and 60% of right reservoir fluid at beginning and reversed at end; aliquoting structure of other mixing aπay should provide a way to preserve this gradient. Each fluid reservoir is fluidly connected with a capillary channel 803 or 804 to capillary junction 805. Each capillary channel is about 0.5mm deep and having a cross- sectional diameter of about 0.5mm and that extends about 5cm. Capillary junction 805 has a depth in the platform surface of about 0.75mm that is greater than the depth of capillaries 803 to 804. Alternatively, capillaries 803 or 804 comprise a sacrificial valve
812. Use of said sacrificial valves can be used in addition to or in place of capillary junction 805.
The fluid components of the mixing aπay are also connected with air channels 810, that have dimensions of about 0.25mm deep and permit venting of air displaced by fluid movement on the platform. In addition, capillary junctions 811 that are about
0.75mm deep are present in the air channels to prevent fluid backflow into the air channel.
Capillary junction 805 is fluidly connected by a naπow capillary channel 806 to mixing chamber 807, which is fluidly connected with channel 808, which is further connected with mixed fluid receiving chamber 809. Alternatively, capillary 806 comprises a sacrificial valve 812. Capillary channel 806 is about 0.5mm deep and having a cross-sectional diameter of from about 0.5mm and that extends from about 0.2cm to about 30cm. Mixing chamber 807 is about 0.75mm deep and having a cross- sectional diameter of from about 0.75mm, and is positioned from about 0.2cm to about 30cm from the center of rotation. Capillary channel 808 is about 0.5mm deep and has a cross-sectional diameter of about 5mm and that extends from about 0.2cm to about 30cm. Capillary channel 806 and capillary channel 808 may be offset in their connection with the mixing chamber as previously described in Example 5, or they can be positioned in the mixing chamber at any convenient position, and Coriolis forces have be relied upon to facilitate mixing.
Capillary 808 is fluidly connected with mixed fluid receiving chamber 809. Mixed fluid receiving chamber 809 is about 0.75mm deep and having a cross-sectional diameter of about 5mm from about, and is positioned from about lcm to about 30cm from the center of rotation. As illustrated in Figures 24A through 24E, in the use of this platform a volume
(ranging from 5-45μL of fluid) of each of the fluids to be mixed is applied to the fluid reservoirs 801 and 802 (Figure 24A). Fluid enters the each of the capillaries 803 and 804 and stops at capillary junction 805. Alternatively, the platforms of the invention are provided containing the fluids to be mixed already in fluid reservoirs 801 and 802. In these embodiments, it is prefeπed that sacrificial valves 812 be provided in capillaries 803 and 804, to prevent evaporation, wetting or leakage of fluid from the reservoirs prior to use.
At a first rotational speed f, of lOOrpm, the fluids from each capillary flows past capillary junction 805 and through mixing chamber 807 (Figures 24B and 24C). In embodiments comprising a sacrificial valve 812, the valve prevents fluid flow into channels 803 and 804. Upon release of sacrificial valve 812, fluid flow proceeds from capillary junction 805 through channel 806 and into mixing chamber 807 (Figure 24D). Fluid flow within mixing chamber 807 is turbulent, in contrast to fluid flow through capillary baπier 805 or channel 806, which is primarily laminar, so that mixing occurs predominantly in mixing chamber 807. Fluid flow proceed through channel 808 and the mixed fluid solution is displaced into mixed fluid receiving chamber 809 (Figures 24D and 24E).
EXAMPLE 8 Immunoassays A microsystems platform provided by the invention and specifically designed for performing immunoassay is illustrated in Figure 25. In the Figure, the aπangement of one assay aπay 19 on a disk 11 is shown; a multiplicity of such aπays can be advantageously aπanged on a microsystems platform, most preferably a disk, of the invention, to provide a multi-use or multi-assay platform. The components of the mixing aπay are shown in greater detail in Figure 26.
Disk embodiments of the platforms of the invention were fashioned from machined acrylic. The overall disc dimensions include an outer radius of about 6cm and an inner radius of about 0.75cm, wherein the disk is mounted on the spindle of a rotary device. The thickness of the disc ranged from about 0.9mm to about 1.5mm. The working fluid volume for reaction was about 10μL.
The components of the mixing aπay were as follows. An entry port 901 having a depth in the platform surface of about 0.75m and lateral dimensions of about 0.5cm was constructed on the platform, and designed to accommodate a volume of about 1 OμL, ranging from 2μL to 20μL. This entry port was fluidly connected with a metering capillary 902 having cross-sectional diameter of about 0.5mm and having a depth of about 0.75mm; the length of this metering capillary was sufficient to contain a total volume of about lOμL. The metering capillary 902 was fluidly connected to capillary junction 904.
The entry port was also constructed to be fluidly connected with an overflow capillary 903 having a cross-sectional diameter of about 0.5mm and proximal ends rounded with respect to entry port 901. The overflow capillary was fluidly connected with an overflow chamber 905 having a depth in the platform surface of about 0.75mm, greater than the depth of the overflow capillary 903. Each of the overflow and fluid chambers was also connected with air ports or air channels, such as 523, that have dimensions of about 0.25mm deep and permit venting of air displaced by fluid movement on the platform. Capillary junctions 524 that were about 0.75mm deep were present in the air channels to prevent fluid flow into the air channels.
Entry port 901 was positioned on the platform 1.3cm from the center of rotation. Metering capillary 902 extends 4.2cm from entry port 9015. Overflow capillary 902 extends from entry port 901 from about lcm to about 20cm. The extent of the length of overflow capillary 903 was 20%> longer than metering capillary 902. The position of overflow chamber 905 was from about lcm to about 20cm from the center of rotation, and the position of capillary junction 904 was 5cm from the center of rotation.
Capillary junction 904 was fluidly connected with capillary channel 906, which in turn was fluidly connected with incubation chamber 910. Capillary channel 906 has a cross-sectional diameter of about 0.5mm and extends about lcm. Incubation chamber
910 has a depth in the platform surface of about 0.75mm, that was greater than the depth of capillary channel 906. Capillary channel 906 was also fluidly connected with channel 909 through capillary junction 907. Capillary junction 907 was constructed to prevent fluid flow of the sample backwards through the junction and into the wash buffer. Channel 909 has a cross-sectional diameter of about 0.5mm and extends about 5cm.
Capillary junction 907 has a depth in the platform surface of about 0.75mm, greater than the depth of the channel 909 or capillary channel 906. Incubation chamber 910 also contains a specific binding species, most preferably an antibody, specific for a component of the sample. This species was advantageously contained within incubation chamber 910 as a coating on the surface of the chamber, or attached to beads or other carrier within the chamber, or to a functionalized inner surface of the chamber.
Capillary junction 907 was further fluidly connected with wash buffer reservoir 516, having a depth in the platform surface of about 0.75mm and positioned at a distance 3.7cm from the axis of rotation.
Capillary junction 907 was further fluidly connected with reagent capillary 920, which was further fluidly connected with capillary junction 914, which was further fluidly connected with channel 926, and which was fluidly connected with reagent reservoir 917. Reagent capillary 920 has a cross-sectional diameter of about 0.25mm and extended about lcm. Capillary junction 914 has a depth in the platform surface of about 0.25mm and positioned at a distance of about 2.7cm from the axis of rotation. Reagent capillary 926 has a cross-sectional diameter of about 0.25mm and extends from about 0.2cm to about 20cm. Reagent reservoir 917 has a depth in the platform surface of about 0.75mm and positioned at a distance of about 2.3cm from the axis of rotation.
Incubation chamber 910 was fluidly connected at a point most distal to the axis of rotation to U-shaped capillary 921. U-shaped capillary 921 has a cross-sectional diameter of about 0.5mm and extended about lcm. This capillary extends in a U-shape to a point that was at least as proximal to the axis of rotation than the most axis-proximal extent of incubation chamber 910. This positioning of the U-shaped channel relative to incubation chamber 910 ensures that additional fluids flowing into incubation chamber 910 and displacing fluid therefrom will displace said fluid homogeneously, i.e., the first fluid in the chamber will be pushed out of the chamber whilst being replaced by the second fluid.
This U-shaped capillary was also fluidly connected with waste reservoir 915. Waste reservoir 915 has a depth in the platform surface of about 0.75mm and positioned at a distance of about 4.5-5.7cm from the axis of rotation. In certain embodiments of the invention, sacrificial valves 922 can be positioned at the junction of capillary junction 904 and capillary channel 906, at the junction of capillary junction 907 and wash buffer capillary 908, or at the junction of reagent reservoir 918 and capillary junction 919.
As illustrated in Figures 27A through 27L, in the use of this platform reagent reservoir 916 and wash reservoir 915 were pre-loaded on the disk, and most preferably the disk contains sacrificial valves 922 at the junction of capillary junction 907 and wash buffer capillary 908, and at the junction of reagent reservoir 918 and capillary junction
919. An imprecise volume (ranging from 1-150μL of fluid) of a fluid was applied to the entry port 901 (Figure 27A). Fluid wicks into metering capillary 902 and stops at the capillary junction between metering capillary 902 and capillary junction 904 (Figures 27B and 27C). After sample loading by a user and filling of metering capillary 902 and overflow capillary 903 at no rotational speed, the platform was spun at a first rotational speed f, of 45rpm.
Due to the greater distance of the end of overflow capillary 903 from the center of rotation than the end of metering capillary 902, fluid flows through overflow capillary 903 into overflow chamber 905 (Figure 27D). The platform was spun until all excess fluid was evacuated from entry port 901 and into overflow chamber 905, except the fluid contained in metering capillary 902 (Figure 27E).
At a second rotational speed f2 of 65rpm, the capillary junction 904 at the distal end of the metering capillary 902 was overcome, and sample from metering capillary 902 fills incubation chamber 910 (Figures 27F and 27G). A portion of the sample wicks into U-shaped capillary 914 to the level of the sample in incubation chamber 910 (Figure
27G). The sample was incubated for a time sufficient for maximum saturation binding of the component in the sample that specifically bind to the specific binding species. At a third rotational speed f, of 450rpm, the capillary junction 908 was overcome, and wash buffer from reservoir 916 flows through capillary 909, capillary
906, and into incubation chamber 910. Wash buffer fluid flow forces the sample through
U-shaped capillary 914 and into waste reservoir 915 (Figures 27 H through 27 J).
Preferably, sacrificial valves 922 were released to permit wash buffer fluid flow.
At a fourth rotational speed f4 of 500rpm, the capillary junction 919 was overcome, and reagent buffer from reservoir 917 flows through capillary 918, capillary
920, capillary junction 908, capillary 906, and into incubation chamber 910. Reagent buffer fluid flow forces the wash buffer through U-shaped capillary 914 and into waste reservoir 515 (Figures 27K through 27L). Preferably, sacrificial valves 922 were released to permit reagent buffer fluid flow.
The reagent buffer contained a chromogen or other developing agent for detection of specific binding in incubation chamber 910. The extent of specific immunoassay being was determined relative to the amount of chromogen produced in the assay.
EXAMPLE 9 Immunoassavs
Use of nitrocellulose for immobilizing antigens for use with the immunoassay aπays described in Example 8 were demonstrated as follows.
Proof-of Principle Visual blue signals have been developed resulting from formation of a three- membered sandwich immobilized on a nitrocellulose (NC) surface. This sandwich is composed of (a) a capture anti-TSH monoclonal antibody (MAB) adsorbed onto porous NC; (b) TSH antigen, and c) a complimentary anti-TSH MAB coated onto colloidal blue latex particles. The intensity of blue color immobilized at the capture site increases in a regular way with antigen (TSH) level, thus providing a method of the quantitative analysis of unknown specimens.
Capture MAB was immobilized on 8-μm NC by application of 2 μL spots at a concentration of 10 mg/mL. These spots spread to circles about " in diameter. After brief incubation, the field surface of the NC was blocked in a bath of 1%> BSA in PBS, followed by extensive washing with 0.1 % BSA-PBS. The membrane was allowed to air dry prior to use. The spotted region was then cut into small squares and introduced into glass test tubes followed by 200 μL aliquots of serially diluted TSH standards in 50/50 horse serum PBS-BSA buffer. After a brief incubation to wet the NC, 10 μL of a 0.309 micron diameter blue latex suspension coated with a complimentary MAB was added and mixed. After 5 min of gentle shaking, the disk was removed, washed under a PBS-BSA stream, and allowed to dry. By visual inspection, the color intensity was seen to vary in the expected way with TSH concentration.
Additional antibody labeled groups, such a colloidal gold, fluorescent or colored latex beads, can be used to detect antibody binding and accumulation.
Experimental: Competitive Immunoassay
A competitive assay for the monovalent thyroid hormone Thyroxine has been developed analogous to and complimenting the sandwich assay described above for TSH. Antigen was immobilized on 5μm NC by application of 1.5 μL volumes of BS A- T4 (12.5 mg/mL) followed by blocking with 1 % BSA in PBS and final washing with 0.1 % BSA in PBS. Anti-thyroxine monoclonal antibody (MAB) was immobilized on
Seradyne 0.309 μm blue latex beads by passive adsorption.
Antibody-coated beads were briefly incubated in test tubes with T4 in buffer over a wide hormone concentration range. Square pieces of nitrocellulose containing adsorbed capture antigen were then added to the test tubes and incubated for 10 minutes, during which time color developed on the NC. The squares were then washed with a small volume of buffer and color intensity visually noted as a function of T4 level.
A roughly monotonic increase in blue color intensity with decreasing T4 concentration was found, as expected for a competitive assay. Change in color intensity occuπed over a low T4 concentration range, implying that a serum specimen can be diluted by about ten-fold for analysis; i.e., about 30μL of serum per test can be used, diluted to 300 μL to produce detectable signal variation over the range of clinical diagnostic interest. A total assay time of about 20-?? minutes was determined without optimization, and low background and high signal intensity were obtained (without optimization). These results indicate that these procedures make it possible to detect immunoassay results at antigen and antibody concentrations useful for the detection of small amounts of antigen in complex mixtures, such as biological mixtures.
EXAMPLE 10 Preparation of Resistive Heater Elements Resistive heater elements were prepared on the platforms of the invention as follows. Portions of the surface of a microsystems platform on which a resistive heater element are desired are screen printed with a resistive ink electrically coupled with a conductive ink to conduct electricity across the resistive ink in the presence of a direct cuπent (DC) voltage drop across the extent of the resistive ink. Both the resistive ink and the conductive ink are screened printed using methods well known in the art (Gilleo, ibid.)
Briefly, a conductive ink pattern, such as those illustrated in Figure 30, was screened onto a heat stabilized polyester sheet substrate (ICI ST505), and cured at 110- 120°C for 10 minutes. A resistive ink was then screened over the cured conductive ink pattern and the composite cured for an additional 10 min at 110-120°C. Typically, the ink is laid down in a layer that is about 10 microns thick, over an area at least 0.5mm x
0.5mm (0.25mm2). However, film patterns of greater thickness have been produced by printing, curing and reprinting the same polyester sheet in the same pattern up to three times. It was found that these thicker films had reduced resistance and increased heating ability at the same applied voltage. In some instances, polymer thick films are covered with a dielectric layer that insulates the electrical circuit; this is particularly advantageous in embodiments of the resistive heaters that are used to heat liquids on the platforms of the invention. This dielectric layer is also deposited by screen printing and then cured using ultraviolet radiation for several seconds at 1000mJ/cm2 . Figures 30 and 31 show an example of a circuit that was screen printed onto heat stabilized polyester. The circuit materials consisted of Dupont 5028 conductive ink and Dupont 7082 resistive ink; the resistive ink comprised the solid back patterned areas and the conductive inks comprised the light, thin lined patterns on the disc. Electrical connection to aDC power source are labeled as 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, and 22. Resistances of the different circuit elements ranged from approximately 10 to approximately 200 Ohms, depending on circuit geometry. For example, a resistance of 12.1 ± 1.2 Ohm (N = 10) was measured between electrical contacts 11 and 12. In another example, a resistance of 1808 ± 257 Ohm (N = 10) was measured between electrical contacts 11 and 22. In another experiment, Dupont 7082 resistive ink (400 Ohm per square per mil) was blended with Dupont 7102 resistive ink (20 Ohm per square per mil). In a 50 : 50 mixture (by wet weight) and then screen printed into the patterns shown in Figures 30 and 31, a resistance of 7.3 ± 0.6 Ohm (N= 1) was measured between electrical contacts 11 and 12. In another example, a resistance of 272.2 ± 22.7 Ohm (N= 7) was measured between electrical contacts 11 and 22. When resistive heater elements were further screen printed, cured, reprinted and re-cured over the same region with Dupont 7082 resistive ink, the resistive circuits were thickened and the resistance was decreased. In this experiment, a second and third printing reduced the resistance between electrical contacts 11 and 12 to 679 ± 86.5 Ohm (N=8). Thus, the ability to tailor resistance through appropriate choice of ink formulation and reprinting of resistive circuits enables control of the electrical properties of the final screen printed circuit.
EXAMPLE 11 Use of Resistive Polymer Thick Films as Heater Elements The use of resistive polymer thick films as heater elements depends on the capacity of resistive polymer thick films to produce heat when an electrical potential is applied across a resistive polymer thick film element produced as described in Example 10. The capacity of a resistive polymer thick film element to be used as a heater is shown in Figure 32. Circuits in the pattern shown in Figures 30 and 31 were screen- printed using Dupont 5028 silver paste as the conductive electrode combined with a 50
: 50 mixture of Dupont 7082 resistive ink blended with Dupont 7102 resistive ink to form the resistive heating element. A direct cuπent (DC) voltage was applied across the electrical contacts 31 and 33 in Figure 31 and the temperature at the surface of the heater measure using a thermistor probe. These experiments were performed "dry", that is, on a resistive heater element without application of a fluid on the platform, and stationary, that is, without rotation of the disc. Figure 32 is a graph of the temperature produced as a function of time and applied (DC) voltage (voltage applied was 2V, 3 V, 4V, 5 V, and 6V, read bottom to top from the voltage that produced the lowest steady-state voltage through the highest steady-state voltage). This graph shows that the maximum temperature produced increased with both time and applied voltage, and that the maximum voltage reached a steady state within between 100-150 seconds, and that the
!1 maximum voltage achieved was about 85-90 °C. These data have been converted into the graph shown in Figure 33, which show a plot of the steady-state temperature obtained as a function of applied (DC) voltage. These results show that the steady-state temperature achieved increases in a parabolic fashion with increased applied voltage. The dependence of maximum steady-state temperature on voltage was also determined using positive temperature coefficient (PTC) inks, specifically Dupont 7285 ink, in experiments performed as described above. The voltage dependence on temperature is linear over the voltage range tested, and the applied (DC) voltages were about ten-fold higher than the results obtained using blended resistive inks. These results are contrasted in Figure 33 with the results obtained with PTC inks, which approach a steady state temperature with increasing voltage, as expected for a positive temperature controlled ink. The results obtained with PTC inks were obtained using a resistive heating element screen-printed in a patterns as shown in Figure 31 using Dupont 7285 PTC ink. The temperature of the substrate at increasing distance from the resistive heater elements of the invention was determined. These results are shown in Figure 34, where the temperature of the disk drops to ambient within 1 -2mm of the resistive heater. These results indicate that resistive heaters can be aπayed in close proximity without the activation of one of the heaters affecting either the adjacent heater or the platform component (such as a sacrificial valve) controlled by the adjacent heater.
These results demonstrate the capacity of resistive elements prepared using polymer thick films according to the teachings of this invention to be used as resistive heater elements on synthetic substrates appropriate for the production of microsystems platforms of this invention.
EXAMPLE 12 Use of Resistive Heaters with Heat- Activated Valves
Resistive heater elements prepared using resistive polymer thick films as described in Examples 10 and 11 are advantageously used to activate heat-activated valves. Heat-activated valves include valves prepared by depositing a "wax" in a channel or capillary of the fluidics structures disclosed herein.
In preparing heat-activated wax valves, a small amount of molten wax is taken up in a preheated plastic pipette sip, and when the tip is applied to a capillary channel, the channel takes up some of the molten wax by capillary action. The wax valve forms when the wax cools and solidifies in the channel or capillary. Examples of particular hydrocarbons useful in preparing wax valves include the monodisperse alkanes eicosane (Tm = 36.8 °C), tetracosane (Tm = 54.0°C), and octocosane (Tm = 64.5 °C), as well as polydisperse waxes such as paraffin (Tm = 54.4°C). Experiments using these different waxes showed that paraffin is prefeπed over the monodisperse hydrocarbons because it is easier to work. Use of temperature-controlled dispensing tips could avoid this distinction and permit monodisperse hydrocarbons to be advantageously used as wax valves.
After production of wax valves as described above, a resistive heater layer is prepared on a polyester substrate and bonded to the platform substrate with tape. Heater positioning is important to ensure that the wax plug is fully melted, is capable of even flow and will not re-crystallize and clog the channel or capillary downstream of the location of the resistive heater.
In an alternative embodiment, the channels and capillaries of the invention comprising wax valves also comprise wax recrystallization chambers, illustrated in Figures 35 and 37, positioned "downstream" (that is, more proximal to the edge of the platform) from the resistive heater. When the disc is assembled and spinning, the melted wax flows into the recrystallization chamber and solidifies, typically on the walls of the recrystallization chamber. It has been found that optimal functioning of such wax valves requires that the resistive heating element be positioned over the entire extent of the wax valve and over at least the interior 25% of the wax recrystallization chamber.
Figures 36 A through 36C provide an illustration of how a platform comprising the wax valves, fluidics structures and resistive polymer thick films is prepared. As shown in Figure 36 A, heater circuits are screen printed on polyester films bonded to a polycarbonate disc with double-sided tape. Fluidics structures are machined onto an acrylic disc as described herein and shown in Figure 36B. Wax valves positioned manually at appropriate regions of the fluidics channels and capillaries of the platforms of the invention. These two components are then bonded with double-sided tape to form the assembled disc (shown in Figure 36C)..
Figure 37 is a more detailed and expanded view of one of the fluid handling aπays of the invention. The screen-printed lead, prepared according to Example 10, is placed on the platform so that the wax valve is completely covered with the screen printed heater element, which also extends over a portion of the wax recrystallization chamber.
Figure 38 illustrates how the heat-activated wax valves can be used to sequentially open the wax valves and permit fluid flow from a fluid chamber into a fluid receptacle. Aqueous dye was loaded into each of the fluid chambers shown in Figure
38. The disc was rotated at about 700 rpm, a speed above the capillary burst rpm. The
Figure illustrates that sequential application of 10V DC across these resistive heaters is sufficient to completely melt the wax valves without reclosure of the wax valves. Also, the heating and melting of any one of these wax valves was insufficient to cause melting of any of the adjacent wax valves, which are separated on the platform by aboutl -2mm.
EXAMPLE 13 Use of Resistive Heaters with Heat- Activated Valves: First Alternative
In an alternative embodiment, heat-recoverable polymers were used as heat activated valves. In this experiment, heat recoverable tubing FP301H (obtained from
3M, Minneapolis, MN) was cut into a sheet and placed in a capillary channel on a microsystems platform of the invention. This capillary channel divided two fluid chambers, the interior-most one of which contained 25-50μL of fluid. The heat- recoverable polymer functioned as a fluid-tight valve, and no leakage of fluid was observed into the second fluid chamber. The disc was then heated to approximately
100°C and the heat recoverable tubing was observed to shrink by approximately 50%, and liquid was observed to be transported through the channel to the second, exterior fluid chamber.
These results demonstrate that heat-recoverable polymers can be advantageously used as a sacrificial valve. A particular advantage of this type of valve when compared with a wax valve is that the polymer sheet is macroscopic object that is retained by the channel, most likely without clogging either the channel or any microfluidics structure downstream from the sacrificial valve site.
EXAMPLE 14 Use of Resistive Heaters with Heat-Activated Valves: Second Alternative
The use of the resistive heater itself as a sacrificial valve was demonstrated. It was observed that certain configurations of the resistive heaters of the invention were capable of melting the platform substrate when sufficient voltages were placed across the resistive heater. In this experiment, 15V (DC) was applied across the heaters shown in Figure 31. After less than 1 second at this voltage, a hole developed within the resistive element itself, typically at its center, having an area of about lmm2. These results suggested that the heater could be used as a sacrificial valve. In this embodiment, a platform comprising a chamber is prepared connected to a channel that terminates at a radius R from the center of rotation. This terminal part of the channel is then bonded to a heater circuit positioned just below the channel termination point. A second fluidics disk is prepared having a channel positioned at precisely the termination point of the first channel, and is then bonded to this heater circuit so that the beginning point of this second channel is positioned just below the resistive element. After bonding the platform layers together, the resistive element will be positioned between the two channels in different layers of the platform, so that heating through the element will conjoin the two channels and permit fluid flow from one chamber to the other at a different layer of the platform.
EXAMPLE 14 Use of Resistive Heaters to Heat a Fluid
The resistive heaters of the invention are also used to heat a fluid in an incubation chamber or other microfluidics component of a platform of the invention. A DC voltage of 40V was applied between contacts 31 and 33 in a resistive heater produced as shown in Figure 31 , in which a fluid chamber containing about 65μL of water was placed in thermal contact. A thermistor was also placed in the fluid chamber to monitor the temperature of the water, and the platform was rotated at about 500rpm. These results are shown in Figure 39. A steady-state temperature was obtained after about 200 seconds. After about 500 seconds, the heater was turned off while rotation of the platform continued. The results shown in Figure 39 demonstrate that a resistive heater element of the invention was able to heat this water sample to about 50°C and maintain this temperature for almost 10 minutes, and that convective cooling of the spinning platform reduced the temperature to ambient within 1-2 minutes of the resistive heater shut-off.
This experiment also demonstrated that temperature can be rapidly cycled in a rotating platform by sequential activation of a resistive heater element.
EXAMPLE 14 Use of Resistive Heaters as Thermosensors
Because screen-printed PTC inks display a larger change in resistance with temperature than other inks, these inks can be used to measure temperature on a screen- printed circuit.
In this embodiment, the temperature-sensing device is screen-printed identically as a resistive heater element; however, rather than applying a voltage across the element to produce heat, the resistance of the element is sampled to determine whether resistance has increase as a result of heating. In one application, the resistive heater element itself could be used both as a heater and a temperature sensing device. In this use, the heater circuit is coupled to an external circuit that is switched between constant voltage and constant (small) cuπent modes. In the constant voltage mode the element would heat up, and in the constant cuπent mode the voltage drop would be measured and the temperature determined thereby.
In another application, a PTC ink is screen-printed over a portion to a resistive heater element. The resistive heater and PTC ink/theπnosensor elements are joined to external circuitry using different sets of conductive leads; the external circuit delivers voltage to the resistive heater element and a small constant cuπent to the thermosensor element. By measuring the voltage drop in the PTC ink element, the temperature of the resistive element is infeπed. Coordination of these two circuits could be provided so that the thermosensor is used to regulate the temperature of the resistive heating element.
It should be understood that the foregoing disclosure emphasizes certain specific embodiments of the invention and that all modifications or alternatives equivalent thereto are within the spirit and scope of the invention.

Claims

What is claimed is:
1. A microsystem platform comprising a) a rotatable platform, comprising a substrate having a first flat, planar surface and a second flat, planar surface opposite thereto, each surface comprising a center about which the platform is rotated, wherein the first surface comprises in combination b) an entry port comprising a depression in the first surface having a volumetric capacity of about 1 to about 150μL, that is fluidly connected with c) a first metering capillary aπay comprising a multiplicity of metering capillaries, and an overflow capillary, each being fluidly connected with the entry port, wherein each capillary defines a cross-sectional area of about 0.02mm to about lmm in diameter, and wherein each capillary extends radially from the center of the platform and defines a first end proximally aπayed towards the center of the platform and a second end distally aπayed from the center of the platform, wherein the proximal end of each capillary defines a curved opening; wherein the first metering capillary aπay defines a volume of the fluid and wherein the first metering capillary aπay is fluidly connected with d) a first fluid chamber having a depth in the surface of the platform equal to or greater than the metering capillary and positioned radially more distant from the center of the platform than the entry port, and wherein the overflow capillary is fluidly connected with e) an overflow chamber having a depth in the surface of the platform equal to or greater than the overflow capillary and positioned radially more distant from the center of the platform than the holding channel and the entry port, wherein a capillary junction is formed at the junction of each of the metering capillaries comprising the metering capillary aπay and the first fluid chamber and at the junction of the overflow capillary and the overflow chamber, whereby fluid placed onto the disk at the entry port flows by capillary action to the junction of each of the metering capillaries comprising the metering capillary aπay and the first fluid chamber, and excess fluid flows by capillary action to the junction of the overflow capillary and the overflow chamber; and wherein rotation of the platform at a first rotation speed motivates fluid displacement in the overflow capillary into the overflow chamber but not fluid displacement in any of the metering capillaries comprising the metering capillary aπay, whereby rotation of the platform at the first rotational speed drains the fluid from the entry port into the overflow chamber; and wherein rotation of the platform at a second rotation speed that is greater than the first rotational speed motivates fluid displacement of the volume of the fluid in the metering capillary aπay into the first fluid chamber; and wherein each of the first fluid chamber and overflow chamber also comprise air displacement channels whereby air displaced by fluid movement is vented to the surface of the platform.
2. The microsystem platform of Claim 1 further comprising
6. a capillary having a first end fluidly connected to the first fluid chamber and having a second end fluidly connected to a holding chamber, having a depth in the surface of the platform equal to or greater than the capillary and positioned radially more distant from the center of the platform than the first fluid chamber, wherein a capillary junction is formed at the junction of the capillary and the holding chamber, whereby fluid in the first fluid chamber flows through the capillary to the junction of the capillary and the holding chamber; and wherein rotation of the platform at a third rotational speed that is greater than the second rotational speed motivates fluid displacement of the volume of the fluid in the first fluid chamber into the holding chamber and wherein the holding chamber also comprises air displacement channels whereby air displaced by fluid movement is vented to the surface of the platform.
3. The microsystem platform of Claim 2 further comprising
1. a capillary having a first end fluidly connected to the holding chamber and having a second end fluidly connected to a read chamber, having a depth in the surface of the platform equal to or greater than the capillary and positioned radially more distant from the center of the platform than the holding chamber, wherein a capillary junction is formed at the junction of the capillary and the read chamber, whereby fluid in the holding chamber flows through the capillary to the junction of the capillary and the read chamber; and wherein rotation of the platform at a fourth rotational speed that is greater than the third rotational speed motivates fluid displacement of the volume of the fluid in the holding chamber into the read chamber and wherein each of the holding chamber and the read chamber also comprises air displacement channels whereby air displaced by fluid movement is vented to the surface of the platform..
4. The microsystem platform of Claim 3, further comprising b) a sacrificial valve in the capillary extending from the holding chamber to the read chamber, wherein release of the sacrificial valve permits fluid flow from the holding chamber to the read chamber at a non-zero rotational speed.
5. The microsystem platform of Claim 4 wherein the sacrificial valve is a solid, semi-solid or viscous liquid hydrocarbon, or a plastic.
6. The microsystem platform of Claim 5 further comprising a heating element in the platform in thermal contact with the sacrificial valve, wherein heating the heating element releases the sacrificial valve.
7. The microsystem platform of Claim 3, wherein the holding chamber comprises a first component of a biological detection assay and the read chamber comprises a second component of a biological detection assay, wherein a sample is assayed for the presence of an analyte.
8. The microsystem platform of Claim 7, wherein the holding chamber comprises carboxypeptidase and a peptide comprising a D-amino acid at its carboxyl terminus, and the read chamber comprises D-amino acid oxidase, flavine adenine dinucleotide, horseradish peroxidase and a chromogen, and the biological detection assay detects the presence of β-lactam antibiotics in a fluid sample.
9. The microsystem platform of Claim 8 wherein the fluid sample is milk.
10. A method for moving a fluid in a microsystem platform according to Claim 1 , the method comprising the steps of a) applying an amount of a fluid sample comprising a volume of about 1 to about 150μL to the entry port of the rotatable microsystem platform; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port and the overflow capillary into the overflow chamber; c) rotating the platform at a second rotation speed that is greater than the first rotational speed displace a volume of the fluid in the metering capillary into the first fluid chamber.
11. A method for moving a fluid in a microsystem platform according to Claim 2, the method comprising the steps of a) applying an amount of a fluid sample comprising a volume of about 1 to about 150μL to the entry port of the rotatable microsystem platform; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port and the overflow capillary into the overflow chamber; c) rotating the platform at a second rotation speed that is greater than the first rotational speed to displace a volume of the fluid in the metering capillary into the first fluid chamber; and c) rotating the platform at a third rotational speed that is greater than the second rotational speed to displace a volume of the fluid in the first fluid chamber into the holding chamber.
12. A method for moving a fluid in a microsystem platform according to Claim 3, the method comprising the steps of a) applying an amount of a fluid sample comprising a volume of about 1 to about 150μL to the entry port of the rotatable microsystem platform; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port and the overflow capillary into the overflow chamber; c) rotating the platform at a second rotation speed that is greater than the first rotational speed to displace a volume of the fluid in the metering capillary into the first fluid chamber; d) rotating the platform at a third rotational speed that is greater than the second rotational speed to displace a volume of the fluid in the first fluid chamber into the holding chamber; and e) rotating the platform at a fourth rotational speed that is greater than the second rotational speed to displace a volume of the fluid in the holding chamber into the read chamber.
13. A method for detecting an amount of a β-lactam antibiotic in a fluid sample using a microsystem platform according to Claim 8, the method comprising the steps of a) applying an amount of a fluid sample comprising a volume of about 1 to about 150μL to the entry port of the rotatable microsystem platform; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port and the overflow capillary into the overflow chamber; c) rotating the platform at a second rotation speed that is greater than the first rotational speed to displace a volume of the fluid in the metering capillary into the first fluid chamber; d) rotating the platform at a third rotational speed that is greater than the second rotational speed to displace a volume of the fluid in the first fluid chamber into the holding chamber; e) incubating the fluid in the holding chamber for a time and at a temperature sufficient to inhibit carboxypeptidase activity; f) rotating the platform at a fourth rotational speed that is greater than the second rotational speed to displace a volume of the fluid in the holding chamber into the read chamber; g) incubating the fluid in the read chamber for a time and at a temperature sufficient to develop the chromogen; h) detecting the amount of developed chromogen in the read chamber and comparing said amount with the amount produced by a sample that does not contain a β-lactam antibiotic.
14. A microsystem platform comprising a) a rotatable platform, comprising a substrate having a first flat, planar surface and a second flat, planar surface opposite thereto, each surface comprising a center about which the platform is rotated, wherein the first surface comprises in combination b) an entry port comprising a depression in the first surface having a volumetric capacity of about 1 to about 150μL, that is fluidly connected with c) a capillary aπay comprising a multiplicity of capillaries, each capillary being fluidly connected with the entry port, wherein each capillary defines a cross-sectional area of about 0.02mm to about lmm in diameter, and wherein each capillary extends radially from the center of the platform and defines a first end proximally aπayed towards the center of the platform and a second end distally aπayed from the center of the platform, wherein the proximal end of each capillary defines a curved opening; wherein the capillary aπay defines a volume of the fluid and wherein the capillary aπay is fluidly connected with d) a first fluid chamber having a depth in the surface of the platform equal to or greater than the metering capillary and positioned radially more distant from the center of the platform than the entry port, and wherein the first fluid chamber is also fluidly connected with an overflow capillary that is fluidly connected with e) an overflow chamber having a depth in the surface of the platform equal to or greater than the overflow capillary and positioned radially more distant from the center of the platform than the holding channel and the entry port, wherein the overflow capillary is fluidly connected with the first fluid chamber at a position on the platform that is closer to the axis of rotation than the position on the platform where the capillaries of the capillary aπay are fluidly connected with the first fluid chamber; wherein a capillary junction is formed at the junction of each of the capillaries comprising the capillary aπay and the first fluid chamber and at the junction of the overflow capillary and the overflow chamber, whereby fluid placed onto the disk at the entry port flows by capillary action to the junction of the each of the capillaries comprising the capillary aπay and the first fluid chamber, and wherein rotation of the platform at a first rotation speed motivates fluid displacement of the volume of the fluid in entry port through the capillary aπay into the first fluid chamber until the level of the fluid in the first fluid chamber reaches the position of the fluid connection between the first fluid chamber and the overflow capillary, wherein excess fluid flows through the overflow capillary to the capillary junction with the overflow chamber at the first rotational speed, and drains excess fluid from the first fluid chamber through the overflow channel and into the overflow chamber, until the level of fluid in the first fluid chamber is decreased to a level farther from the center of rotation in the first fluid chamber than the position where the overflow capillary is fluidly connected with the chamber, thereby leaving a defined volume of fluid in the first fluid chamber; and wherein each of the first fluid chamber and overflow chamber also comprise air displacement channels whereby air displaced by fluid movement is vented to the surface of the platform.
15. The microsystem platform of Claim 14 further comprising f) a second capillary having a first end fluidly connected to the first fluid chamber and having a second end fluidly connected to a holding chamber, having a depth in the surface of the platform equal to or greater than the capillary and positioned radially more distant from the center of the platform than the first fluid chamber, wherein a capillary junction is formed at the junction of the capillary and the holding chamber, whereby fluid in the first fluid chamber flows through the capillary to the junction of the capillary and the holding chamber during rotation at the first rotational speed; and wherein rotation of the platform at a second rotational speed that is greater than the first rotational speed motivates fluid displacement of the volume of the fluid in the first fluid chamber into the holding chamber and wherein the holding chamber also comprises air displacement channels whereby air displaced by fluid movement is vented to the surface of the platform.
16. The microsystem platform of Claim 15 further comprising g) a capillary having a first end fluidly connected to the holding chamber and having a second end fluidly connected to a read chamber, having a depth in the surface of the platform equal to or greater than the capillary and positioned radially more distant from the center of the platform than the holding chamber, wherein a capillary junction is formed at the junction of the capillary and the read chamber, whereby fluid in the holding chamber flows through the capillary to the junction of the capillary and the read chamber; and wherein rotation of the platform at a third rotational speed that is greater than the second rotational speed motivates fluid displacement of the volume of the fluid in the holding chamber into the read chamber and wherein each of the holding chamber and the read chamber also comprises air displacement channels whereby air displaced by fluid movement is vented to the surface of the platform.
17. The microsystem platform of Claim 16, further comprising h) a sacrificial valve in the capillary extending from the holding chamber to the read chamber, wherein release of the sacrificial valve permits fluid flow from the holding chamber to the read chamber at a non-zero rotational speed.
18. The microsystem platform of Claim 17 wherein the sacrificial valve is a solid, semi-solid or viscous liquid hydrocarbon, or a plastic.
19. The microsystem platform of Claim 18 further comprising a heating element in the platform in thermal contact with the sacrificial valve, wherein heating the heating element releases the sacrificial valve.
20. The microsystem platform of Claim 16, wherein the holding chamber comprises a first component of a biological detection assay and the read chamber comprises a second component of a biological detection assay, wherein a sample is assayed for the presence of an analyte.
21. The microsystem platform of Claim 20, wherein the holding chamber comprises carboxypeptidase and a peptide comprising a D-amino acid at its carboxyl terminus, and the read chamber comprises D-amino acid oxidase, flavine adenine dinucleotide, horseradish peroxidase and a chromogen, and the biological detection assay detects the presence of β-lactam antibiotics in a fluid sample.
22. The microsystem platform of Claim 21 wherein the fluid sample is milk.
23. A method for moving a fluid in a microsystem platform according to Claim 14, the method comprising the steps of a) applying an amount of a fluid sample comprising a volume of about 1 to about 150μL to the entry port of the rotatable microsystem platform; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port and the capillary aπay into the first fluid chamber and the overflow capillary; and c) rotating the platform at a said first rotational speed for a time sufficient to displace an excess volume of the fluid in the first fluid chamber into the overflow chamber.
24. A method for moving a fluid in a microsystem platform according to Claim 15, the method comprising the steps of a) applying an amount of a fluid sample comprising a volume of about 1 to about 150μL to the entry port of the rotatable microsystem platform; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port and the capillary aπay into the first fluid chamber, the second capillary and the overflow capillary; c) rotating the platform at a said first rotational speed for a time sufficient to displace an excess volume of the fluid in the first fluid chamber into the overflow chamber; and d) rotating the platform at a third rotational speed that is greater than the second rotational speed to displace a volume of the fluid in the first fluid chamber into the holding chamber.
25. A method for moving a fluid in a microsystem platform according to Claim 16, the method comprising the steps of a) applying an amount of a fluid sample comprising a volume of about 1 to about 150μL to the entry port of the rotatable microsystem platform; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port and the capillary aπay into the first fluid chamber, the second capillary and the overflow capillary; c) rotating the platform at a said first rotational speed for a time sufficient to displace an excess volume of the fluid in the first fluid chamber into the overflow chamber; d) rotating the platform at a second rotational speed that is greater than the first rotational speed to displace a volume of the fluid in the first fluid chamber into the holding chamber; and e) rotating the platform at a third rotational speed that is greater than the second rotational speed to displace a volume of the fluid in the holding chamber into the read chamber.
26. A method for detecting an amount of a β-lactam antibiotic in a fluid sample using a microsystem platform according to Claim 21 , the method comprising the steps of a) applying an amount of a fluid sample comprising a volume of about 1 to about 150μL to the entry port of the rotatable microsystem platform; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port and the capillary aπay into the first fluid chamber, the second capillary and the overflow capillary; c) rotating the platform at a said first rotational speed for a time sufficient to displace an excess volume of the fluid in the first fluid chamber into the overflow chamber; and d) rotating the platform at a second rotational speed that is greater than the first rotational speed to displace a volume of the fluid in the first fluid chamber into the holding chamber; e) incubating the fluid in the holding chamber for a time and at a temperature sufficient to inhibit carboxypeptidase activity; f) rotating the platform at a third rotational speed that is greater than the second rotational speed to displace a volume of the fluid in the holding chamber into the read chamber; g) incubating the fluid in the read chamber for a time and at a temperature sufficient to develop the chromogen; h) detecting the amount of developed chromogen in the read chamber and comparing said amount with the amount produced by a sample that does not contain a β-lactam antibiotic.
27. A microsystem platform comprising a) a rotatable platform, comprising a substrate having a first flat, planar surface and a second flat, planar surface opposite thereto, each surface comprising a center about which the platform is rotated, wherein the first surface comprises in combination b) an entry port comprising a depression in the first surface having a volumetric capacity of about 1 to about 50μL, that is fluidly connected with c) an entry capillary that defines a cross-sectional area of about 0.1 to about 2cm in diameter and having a volume of about 5 to 25 μL, wherein the capillary extends radially from the center of the platform and defines a first end proximally aπayed towards the center of the platform and a second end distally aπayed from the center of the platform, wherein the proximal end of each capillary defines a curved opening; wherein the entry capillary is fluidly connected with d) a separation column having a cross-sectional diameter of about 0.1 to about 2cm and having a volume of about 5 to 25 μL, and having a depth in the surface of the platform equal to or greater than the entry capillary, and that defines a first, closed end proximally aπayed towards the center of the platform and a second closed end distally aπayed from the center of the platform, wherein the separation chamber is positioned substantially parallel to the entry capillary and wherein the entry capillary is fluidly connected with the separation chamber at a position on the platform that is substantially more proximal to the second end of the chamber than the first end of the chamber; wherein a capillary junction is formed at the junction of the entry capillary and the separation chamber, whereby fluid placed onto the disk at the entry port flows by capillary action to the junction of the entry capillary and the separation chamber, and wherein rotation of the platform at a first rotation speed motivates fluid displacement in the entry capillary into the separation; and wherein the separation chamber also comprises air displacement channels whereby air displaced by fluid movement is vented to the surface of the platform.
28. The microsystem platform of Claim 27 further comprising e) an overflow chamber fluidly connected to the separation chamber by a channel, wherein the position of the channel on the platform is substantially more proximal to the first end of the chamber than the second end of the chamber; and wherein the overflow chamber also comprises air displacement channels whereby air displaced by fluid movement is vented to the surface of the platform.
29. The microsystem platform of Claim 28 further comprising f) an capillary fluidly connected to the separation chamber at a first end and having a cross-sectional diameter of about 0.05mm to about 0.25mm and a second end fluidly connected with a capillary junction having a depth in the surface of the platform greater than the depth of the capillary, wherein the capillary junction is fluidly connected with g) a decant capillary having a cross-sectional diameter of about 0.25mm to about lmm, wherein the capillary extends radially from the center of the platform and defines a first end proximally aπayed towards the center of the platform and a second end distally aπayed from the center of the platform, wherein the second end of the decant capillary is fluidly connected with h) a decant chamber having a depth in the surface of the platform equal to or greater than the capillary and positioned radially more distant from the center of the platform than the capillary junction, wherein rotation of the platform at a rotational speed greater than the first rotational speed motivates fluid flow through the capillary junction and into the decant chamber, and wherein the decant chamber also comprises air displacement channels whereby air displaced by fluid movement is vented to the surface of the platform.
30. A method for separating a fluid from a suspension of particulate in said fluid using a microsystem platform according to Claim 27, the method comprising the steps of a) applying an amount of a fluid sample comprising a particulate suspension having a volume of about 1 to about 50μL to the entry port of the rotatable microsystem platform and allowing the fluid to fill the entry capillary; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port into the separation chamber; and c) rotating the platform at a second rotation speed that is greater than the first rotational speed for a time sufficient to cause the particulates in the suspension to be concentrated in a portion of the separation chamber that is closer to the second end of the chamber than the first end of the chamber.
31. A method for separating a fluid from a suspension of particulate in said fluid using a microsystem platform according to Claim 28, the method comprising the steps of a) applying an amount of a fluid sample comprising a particulate suspension having a volume of about 1 to about 50μL to the entry port of the rotatable microsystem platform and allowing the fluid to fill the entry capillary; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port into the separation chamber and for excess fluid to be displaced into the overflow chamber; and c) rotating the platform at a second rotation speed that is greater than the first rotational speed for a time sufficient to cause the particulates in the suspension to be concentrated in a portion of the separation chamber that is closer to the second end of the chamber than the first end of the chamber.
32. A method for separating a fluid from a suspension of particulate in said fluid using a microsystem platform according to Claim 29, the method comprising the steps of a) applying an amount of a fluid sample comprising a particulate suspension having a volume of about 1 to about 50μL to the entry port of the rotatable microsystem platform and allowing the fluid to fill the entry capillary; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port into the separation chamber and for excess fluid to be displaced into the overflow chamber; c) rotating the platform at a second rotation speed that is greater than the first rotational speed for a time sufficient to cause the particulates in the suspension to be concentrated in a portion of the separation chamber that is closer to the second end of the chamber than the first end of the chamber; and d) rotating the platform at a third rotational speed that is greater than the second rotational speed to motivate displacement of fluid from the separation column through the capillary junction and the decant capillary and into the decant chamber, wherein the fluid is substantially free of particulates.
33. A method according to Claim 30 wherein the fluid is blood
34. A method according to Claim 31 wherein the fluid is blood
35. A method according to Claim 32 wherein the fluid is blood
36. A microsystem platform of Claim 29 wherein the particulate suspension has a viscosity that prevents fluid flow through the second capillary.
37. A microsystem platform comprising a) a rotatable platform, comprising a substrate having a first flat, planar surface and a second flat, planar surface opposite thereto, each surface comprising a center about which the platform is rotated, wherein the first surface comprises in combination b) an entry port comprising a depression in the first surface having a volumetric capacity of about 5 to about 50μL, that is fluidly connected with c) an overflow capillary that is fluidly connected with d) an overflow chamber having a depth in the surface of the platform equal to or greater than the overflow capillary and positioned radially more distant from the center of the platform than the entry port, and wherein the entry port is fluidly connected with e) a first metering capillary aπay and a metering second capillary array, each comprising a multiplicity of capillaries, each capillary being fluidly connected with the entry port, wherein each capillary defines a cross- sectional area of about 0.02mm to about lmm in diameter, and wherein each capillary extends radially from the center of the platform and defines a first end proximally aπayed towards the center of the platform and a second end distally aπayed from the center of the platform, wherein the proximal end of each capillary defines a curved opening; wherein the capillary aπay defines a volume of the fluid and wherein the first metering capillary aπay is fluidly connected with f) a ballast chamber having a depth in the surface of the platform equal to or greater than the metering capillary and positioned radially more distant from the center of the platform than the entry port but radially less distant that the capillary junction, wherein a capillary junction is formed at the junction of each of the capillaries comprising the second metering capillary array and the ballast chamber; and wherein the second metering capillary aπay is fluidly connected with g) a capillary junction having a depth in the surface of the platform equal to or greater than the metering capillary and positioned radially more distant from the center of the platform than the entry port; and wherein the capillary junction is fluidly connected to h) a channel that is fluidly connected to i) a separation chamber that is positioned radially more distant from the center of the platform than the ballast chamber and having a first end proximally aπayed towards the center of the platform and a second end distally aπayed from the center of the platform; whereby fluid placed onto the disk at the entry port flows by capillary action to the junction of the each of the metering capillaries comprising the first metering capillary array and the ballast chamber, and to the junction of each of the metering capillaries of the second metering capillary aπay and the capillary junction, and excess fluid flows by capillary action to the junction of the overflow capillary and the overflow chamber; and wherein rotation of the platform at a first rotation speed motivates fluid displacement in the overflow capillary into the overflow chamber but not fluid displacement in any of the metering capillaries comprising the first or second metering capillary aπays, whereby rotation of the platform at the first rotational speed drains the fluid from the entry port into the overflow chamber; and wherein rotation of the platform at a second rotation speed that is greater than the first rotational speed motivates fluid displacement of the volume of the fluid in the metering capillaries of the first capillary aπay into the ballast chamber; and displacement of the volume of the fluid in the metering capillaries of the second capillary aπay through the capillary boundary and the channel and into the separation chamber; wherein each of the ballast chamber, the overflow chamber and the separation chamber also comprise air displacement channels whereby air displaced by fluid movement is vented to the surface of the platform.
38. The microsystem platform of Claim 37 further comprising j) a decant channel having a cross-sectional diameter of about 0.5mm to about lmm, wherein the channel extends radially from the center of the platform and having a first end proximally aπayed towards the center of the platform and fluidly connected to the separation chamber at a position on the platform that is substantially more proximal to the first end of the chamber than the second end of the chamber, wherein the second end of the decant capillary is fluidly connected with k) a decant chamber having a depth in the surface of the platform equal to or greater than the capillary and positioned radially more distant from the center of the platform than the separation chamber; wherein the volume of the fluid in the separation chamber delivered from the second metering capillary aπay is insufficient to fill the separation chamber to a level equal to the position on the platform of the fluid connection to the decant channel; wherein the decant chamber also comprises air displacement channels whereby air displaced by fluid movement is vented to the surface of the platform.
39. The microsystem platform of Claim 38 further comprising
1) a capillary having a first end fluidly connected to the ballast chamber and having a second end fluidly connected to a capillary junction having a depth in the surface of the platform equal to or greater than the capillary and positioned radially more distant from the center of the platform than the ballast chamber, and wherein the capillary junction is fluidly connected with m) a channel having a cross-sectional diameter of about 0.5mm to about lmm, wherein the channel extends radially from the center of the platform and having a first end proximally aπayed towards the center of the platform and fluidly connected to the capillary junction, and a second end that is fluidly connected to the separation chamber at a position equal to the position the second end of the separation chamber, wherein rotation of the platform at a third rotational speed that is greater than the second rotational speed motivates fluid displacement of the volume of the fluid in the ballast chamber through the capillary, the capillary junction and the channel and into the separation chamber.
40. The microsystem platform of Claim 39, further comprising n) a sacrificial valve in the capillary extending from the ballast chamber to the separation chamber, wherein release of the sacrificial valve permits fluid flow from the ballast chamber to the separation chamber at a nonzero rotational speed.
41. The microsystem platform of Claim 40 wherein the sacrificial valve is a solid, semi-solid or viscous liquid hydrocarbon, or a plastic.
42. The microsystem platform of Claim 41 further comprising a heating element in the platform in thermal contact with the sacrificial valve, wherein heating the heating element releases the sacrificial valve.
43. A method for separating a fluid from a suspension of particulate in said fluid using a microsystem platform according to Claim 37, the method comprising the steps of a) applying an amount of a fluid sample comprising a particulate suspension having a volume of about 1 to about 50μL to the entry port of the rotatable microsystem platform and allowing the fluid to fill the capillaries of each of the first and second metering capillary aπays and the overflow capillary; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port and the overflow capillary into the overflow chamber; c) rotating the platform at a second rotation speed that is greater than the first rotational speed to displace a volume of the fluid in the first metering capillary into the ballast chamber, and to displace a volume of the fluid from the second metering capillary into the separation chamber; and d) rotating the platform at a third rotation speed that is greater than the first rotational speed for a time sufficient to cause the particulates in the suspension to be concentrated in a portion of the separation chamber that is closer to the second end of the chamber than the first end of the chamber.
44. A method for separating a fluid from a suspension of particulate in said fluid using a microsystem platform according to Claim 39, the method comprising the steps of a) applying an amount of a fluid sample comprising a particulate suspension having a volume of about 1 to about 50μL to the entry port of the rotatable microsystem platform and allowing the fluid to fill the capillaries of each of the first and second metering capillary aπays and the overflow capillary; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port and the overflow capillary into the overflow chamber; c) rotating the platform at a second rotation speed that is greater than the first rotational speed to displace a volume of the fluid in the first metering capillary into the ballast chamber, and to displace a volume of the fluid from the second metering capillary into the separation chamber; d) rotating the platform at a third rotation speed that is greater than the second rotational speed for a time sufficient to cause the particulates in the suspension to be concentrated in a portion of the separation chamber that is closer to the second end of the chamber than the first end of the chamber; e) rotating the platform at a fourth rotational speed that is greater than the third rotational speed to motivate displacement of fluid from the ballast chamber into the separation column, wherein the level of the fluid in the separation column rises to the level of the fluid connection between the separation column and the decant channel, whereby displacement of the volume of the fluid in the ballast chamber into the separation chamber decants an equal volume into the decant chamber, wherein the fluid is substantially free of particulates.
45. A method according to Claim 37 wherein the fluid is blood
46. A method according to Claim 38 wherein the fluid is blood
47. A method according to Claim 39 wherein the fluid is blood.
48. A method according to Claim 44 wherein the fluid is blood.
49. A microsystem platform of Claim 39 wherein the particulate suspension has a viscosity that prevents fluid flow through the second capillary.
50. A microsystem platform comprising a) a rotatable platform, comprising a substrate having a first flat, planar surface and a second flat, planar surface opposite thereto, each surface comprising a center about which the platform is rotated, wherein the first surface comprises in combination b) a first fluid chamber comprising a volume of a first fluid and a second fluid chamber comprising a second volume of a second fluid, wherein the first fluid chamber is fluidly connected to a first capillary and the second fluid chamber is fluidly connected to a second capillary, and wherein each of the first and second capillaries are fluidly connected to c) a capillary junction having a depth in the surface of the platform equal to or greater than the first or second capillaries and positioned radially more distant from the center of the platform than either of the fluid chambers, and wherein the capillary junction is fluidly connected to d) a mixing chamber inlet capillary extending radially on the platform from the capillary junction and being further fluidly connected to e) a mixing chamber having a depth in the surface of the platform equal to or greater than the inlet capillary and positioned radially more distant from the center of the platform than either of the capillary junction, the mixing chamber further comprising a mixing chamber outlet capillary fluidly connected thereto.
51. A microsystem platform according to Claim 50 wherein the volumes contained in the first and second fluid chambers are equal.
52. A microsystem platform according to Claim 50 wherein the volumes contained in the first and second fluid chambers are unequal.
53. A microsystem platform according to Claim 50 wherein the volumes contained in the first and second fluid chambers are mixed to produce a gradient, wherein the shape and position of the first fluid chamber produces a greater fluid volume than the fluid volume displaced from the second fluid chamber when the platform is first rotated at a non-zero rotational speed, and wherein said displaced fluid volume in the first fluid chamber is reduced at a faster rate during rotation than the displaced fluid volume from the second fluid chamber.
54. A microsystems platform according to Claim 50 further comprising e) a mixed fluid receiving chamber having a depth in the surface of the platform equal to or greater than the capillaries and positioned radially more distant from the center of the platform than the mixing chamber.
55. A microsystems platform according to Claim 54, further comprising a multiplicity of mixing chambers arrayed radially across the platform surface, each mixing chamber having an inlet capillary extending radially from a position more proximal to the center of rotation than the mixing chamber, and an outlet capillary extending radially to a position more distal to the center of rotation than the mixing chamber, wherein the inlet capillary of each mixing chamber is fluidly connected with the first mixing chamber of Claim 50 or is the outlet chamber of the mixing chamber immediately more proximal to the center of rotation, and the outlet capillary of each mixing chamber is the inlet capillary of the mixing chamber immediately more distal to the center of rotation, and wherein the outlet capillary of the mixing chamber positioned the most distal from the center of rotation is fluidly connected with a mixed fluid receiving chamber.
56. A microsystems platform according to Claim 53 further comprising a mixed fluid receiving chamber having a depth in the surface of the platform equal to or greater than the capillaries and positioned radially more distant from the center of the platform than the mixing chamber, wherein the mixing chamber comprises a multiplicity of compartments laterally aπayed on the platform, wherein the outlet capillary is fluidically attached to the mixed fluid receiving chamber at a position most proximal to the center of rotation and at the lateral side of the mixing chamber, wherein fluid flow through the capillary and into the mixing chamber sequentially fills each of the compartments of the mixing chamber from the compartment at the closest lateral extent to the fluid connection to the outlet capillary to the compartment at the farthest lateral extent to the fluid connection to the outlet capillary.
57. A method of mixing two fluids using a microsystem platform according to Claim 54, the method comprising the steps of: a) applying a volume of a first fluid to the first fluid chamber and applying a volume of a second fluid to the second fluid chamber; b) rotating the platform at a rotational speed sufficient to motivate fluid flow from the fluid chambers through the capillaries and past the capillary junction and into the mixing chamber, whereby turbulent fluid flow is produced in the mixing chamber; and c) collecting the mixed fluids in the mixed fluid receiving chamber.
58. The method of claim 57 wherein the two fluids differ in the concentration of a solute.
59. The method of claim 57 wherein the two fluids have different viscosities.
60. A microsystem platform comprising a) a rotatable platform, comprising a substrate having a first flat, planar surface and a second flat, planar surface opposite thereto, each surface comprising a center about which the platform is rotated, wherein the first surface comprises in combination b) a first entry port and a second entry port, each comprising a depression in the first surface having a volumetric capacity of about 5 to about 50μL, that is fluidly connected with c) a first capillary aπay and a second capillary aπay, each comprising a multiplicity of capillaries, each capillary of the first capillary aπay being fluidly connected with the first entry port, and each capillary of the second capillary aπay being fluidly connected with the second entry port, wherein each capillary defines a cross-sectional area of about 0.02mm to about lmm in diameter, and wherein each capillary extends radially from the center of the platform and defines a first end proximally aπayed towards the center of the platform and a second end distally aπayed from the center of the platform, wherein the proximal end of each capillary defines a curved opening; wherein the capillary aπay defines a volume of the fluid and wherein the capillary aπays are fluidly connected with d) a curved capillary baπier having a depth in the surface of the platform equal to or greater than the capillary and positioned radially more distant from the center of the platform, wherein a capillary junction is formed at the junction of each of the capillaries comprising the capillary aπays and the curved capillary baπier; whereby a first fluid placed onto the platform at the first entry port flows by capillary action to the junction of the each of the capillaries comprising the first capillary aπay and the curved capillary junction, and a second fluid placed onto the platform at the second entry port flows by capillary action to the junction of the each of the capillaries comprising the second capillary aπay and the curved capillary junction, and wherein rotation of the platform at a first rotation speed motivates fluid displacement of the volume of the fluid in the capillaries of the first and second capillary aπays into the curved capillary junction; and wherein the curved capillary junction also comprise air displacement channels whereby air displaced by fluid movement is vented to the surface of the platform.
61. A microsystems platform according to Claim 60 further comprising: e) a mixing chamber inlet capillary extending radially on the platform from the curved capillary junction and being further fluidly connected to f) a mixing chamber having a depth in the surface of the platform equal to or greater than the inlet capillary and positioned radially more distant from the center of the platform than the capillary junction, the mixing chamber further comprising a mixing chamber outlet capillary fluidly connected thereto.
62. A microsystems platform according to Claim 61 further comprising: g) a mixed fluid receiving chamber having a depth in the surface of the platform equal to or greater than the mixing chamber outlet capillary and positioned radially more distant from the center of the platform than the mixing chamber.
63. A microsystem platform according to Claim 60 wherein the volumes contained in the first and second fluid chambers are equal.
64. A microsystem platform according to Claim 60 wherein the volumes contained in the first and second fluid chambers are unequal.
65. A microsystems platform according to Claim 62, further comprising a multiplicity of mixing chambers aπayed radially across the platform surface, each mixing chamber having an inlet capillary extending radially from a position more proximal to the center of rotation than the mixing chamber, and an outlet capillary extending radially to a position more distal to the center of rotation than the mixing chamber, wherein the inlet capillary of each mixing chamber is fluidly connected with the first mixing chamber of Claim 62 or is the outlet chamber of the mixing chamber immediately more proximal to the center of rotation, and the outlet capillary of each mixing chamber is the inlet capillary of the mixing chamber immediately more distal to the center of rotation, and wherein the outlet capillary of the mixing chamber positioned the most distal from the center of rotation is fluidly connected with a mixed fluid receiving chamber.
66. A microsystems platform according to Claim 61 further comprising a mixed fluid receiving chamber having a depth in the surface of the platform equal to or greater than the capillaries and positioned radially more distant from the center of the platform than the mixing chamber, wherein the mixing chamber comprises a multiplicity of compartments laterally aπayed on the platform, wherein the outlet capillary is fluidically attached to the mixed fluid receiving chamber at a position most proximal to the center of rotation and at the lateral side of the mixing chamber, wherein fluid flow through the capillary and into the mixing chamber sequentially fills each of the compartments of the mixing chamber from the compartment at the closest lateral extent to the fluid connection to the outlet capillary to the compartment at the farthest lateral extent to the fluid connection to the outlet capillary.
68. A method of mixing two fluids using a microsystem platform according to Claim 62, the method comprising the steps of: a) applying a volume of a first fluid to a first entry port and applying a volume of a second fluid to the second entry port and allowing the fluid to fill the first and second capillary aπays by capillary action; b) rotating the platform at a first rotational speed sufficient to motivate fluid flow from the capillary aπays into the curved capillary junction; c) rotating the platform at a second rotational speed that is greater than the fist rotational speed to motivate fluid flow past the capillary junction, through the mixing chamber inlet capillary and into the mixing chamber, whereby turbulent fluid flow is produced in the mixing chamber; and d) collecting the mixed fluids in the mixed fluid receiving chamber.
69. The method of claim 68 wherein the two fluids differ in the concentration of a solute.
70. The method of claim 68 wherein the two fluids have different viscosities.
71. A microsystem platform comprising a) a rotatable platform, comprising a substrate having a first flat, planar surface and a second flat, planar surface opposite thereto, each surface comprising a center about which the platform is rotated, wherein the first surface comprises in combination b) an entry port comprising a depression in the first surface having a volumetric capacity of about 1 to about lOOμL, that is fluidly connected with c) a metering capillary and an overflow capillary, each being fluidly connected with the entry port, wherein each capillary defines a cross- sectional area of about 0.02mm to about lmm in diameter, and wherein each capillary extends radially from the center of the platform and defines a first end proximally aπayed towards the center of the platform and a second end distally aπayed from the center of the platform, wherein the proximal end of each capillary defines a curved opening; wherein the metering capillary defines a volume of the fluid and wherein the metering capillary is fluidly connected with d) a first capillary junction having a depth in the surface of the platform equal to or greater than the metering capillary and positioned radially more distant from the center of the platform than the entry port, and wherein the overflow capillary is fluidly connected with e) an overflow chamber having a depth in the surface of the platform equal to or greater than the overflow capillary and positioned radially more distant from the center of the platform than the holding channel and the entry port, wherein a capillary junction is formed at the junction of the metering capillary and the first capillary junction, and at the junction of the overflow capillary and the overflow chamber, whereby fluid placed onto the disk at the entry port flows by capillary action to the junction of the metering capillary and the first capillary junction, and excess fluid flows by capillary action to the junction of the overflow capillary and the overflow chamber; and wherein rotation of the platform at a first rotation speed motivates fluid displacement in the overflow capillary into the overflow chamber but not fluid displacement in the metering capillary, whereby rotation of the platform at the first rotational speed drains the fluid from the entry port into the overflow chamber; and wherein rotation of the platform at a second rotation speed that is greater than the first rotational speed motivates fluid displacement of the volume of the fluid in the metering capillary past the first capillary junction; and wherein the overflow chamber also comprises air displacement channels whereby air displaced by fluid movement is vented to the surface of the platform.
71. The microsystem platform of Claim 70 further comprising
6. an incubation chamber inlet capillary having a first end fluidly connected to the first capillary junction and having a second end fluidly connected to
7. an incubation chamber, having a depth in the surface of the platform equal to or greater than the capillary and positioned radially more distant from the center of the platform than the first capillary junction whereby fluid in the metering capillary flows past the first capillary junction and through the incubation chamber inlet capillary into the incubation chamber when the platform is rotated at a second rotational speed.
72. The microsystem platform of Claim 71 further comprising f) an incubation chamber outlet capillary having a first end fluidly connected to the incubation chamber at a position most distal from the center of rotation, and having a second end fluidly connected to g) an waste chamber, having a depth in the surface of the platform equal to or greater than the capillary and positioned radially more distant from the center of the platform than the incubation chamber wherein the incubation chamber outlet position extends radially from the position of fluid connection with the incubation chamber substantially parallel to the lateral extent of the incubation chamber, and wherein the capillary comprises a substantially semicircular bend at a position about equal to the side of the incubation chamber most proximal to the center of rotation to produce a pressure block, whereby fluid flow in the outlet capillary is balanced by the volume of the fluid in the incubation chamber, thereby preventing fluid flow into the waste chamber at the second rotational speed.
73. The microsystem platform of Claim 72 further comprising h) a wash buffer reservoir positioned radially more distant from the center of the platform than the entry port but less distant from the center of rotation than the incubation chamber, wherein the wash buffer reservoir is fluidly connected with
I) a wash buffer outlet capillary having a first end fluidly connected to the wash buffer reservoir and having a second end fluidly connected to j) a second capillary junction having a depth in the surface of the platform equal to or greater than the outlet capillary and positioned radially more distant from the center of the platform than the wash buffer reservoir, wherein the capillary junction is fluidly connected to the incubation chamber inlet capillary wherein rotation of the platform at a third rotational speed that is greater than the second rotational speed motivates fluid flow of the wash buffer through the wash buffer outlet capillary, the capillary junction and the incubation inlet capillary and into the incubation chamber, whereby displacement of the volume of the fluid in the incubation chamber by the wash buffer overcomes the pressure block in the incubation chamber outlet capillary, thereby permitting the fluid in the incubation chamber to be displaced by the wash buffer and to flow into the waste chamber at the third rotational speed.
74. The microsystem platform of Claim 73 further comprising k) a reagent reservoir positioned radially less distant from the center of the platform than the incubation chamber, wherein the reagent reservoir is fluidly connected with 1) a reagent outlet capillary having a first end fluidly connected to the reagent reservoir and having a second end fluidly connected to m) the second capillary junction, wherein rotation of the platform at a fourth rotational speed that is greater than the third rotational speed motivates fluid flow of the wash buffer through the reagent reservoir outlet capillary, the capillary junction and the incubation inlet capillary and into the incubation chamber, whereby displacement of the volume of the wash buffer in the incubation chamber by the reagent overcomes the pressure block in the incubation chamber outlet capillary, thereby permitting the fluid in the incubation chamber to be displaced by the reagent and to flow into the waste chamber at the fourth rotational speed.
75. The microsystem platform of Claim 74, further comprising a) a sacrificial valve in the capillary extending from the reagent reservoir to the incubation chamber, wherein release of the sacrificial valve permits fluid flow from the holding chamber to the read chamber at a non-zero rotational speed.
76. The microsystem platform of Claim 75 wherein the sacrificial valve is a solid, semi-solid or viscous liquid hydrocarbon, or a plastic.
77. The microsystem platform of Claim 76 further comprising a heating element in the platform in thermal contact with the sacrificial valve, wherein heating the heating element releases the sacrificial valve.
78. A method for performing an affinity binding assay using a microsystem platform according to Claim 72, the method comprising the steps of a) applying an amount of a fluid sample comprising a first member of an affinity binding pair and having a volume of about 1 to about lOOμL to the entry port of the rotatable microsystem platform and allowing the metering capillary and overflow capillary to fill by capillary action; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port and the overflow capillary into the overflow chamber; c) rotating the platform at a second rotation speed that is greater than the first rotational speed displace a volume of the fluid in the metering capillary into the incubation chamber, wherein the incubation chamber contains a second member of the affinity binding pair; and d) detecting affinity binding in the incubation chamber.
79. A method for performing an affinity binding assay using a microsystem platform according to Claim 73, the method comprising the steps of a) applying an amount of a fluid sample comprising a first member of an affinity binding pair and having a volume of about 1 to about lOOμL to the entry port of the rotatable microsystem platform and allowing the metering capillary and overflow capillary to fill by capillary action; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port and the overflow capillary into the overflow chamber; c) rotating the platform at a second rotational speed that is greater than the first rotational speed displace a volume of the fluid in the metering capillary into the incubation chamber, wherein the incubation chamber contains a second member of the affinity binding pair; d) incubating the fluid in the incubation chamber for a time sufficient to form the affinity binding pair; e) rotating the platform at a third rotational speed that is greater than the second rotational speed to displace a volume of the wash fluid into the incubation chamber; and f) detecting affinity binding in the incubation chamber.
80. A method for performing an affinity binding assay using a microsystem platform according to Claim 74, the method comprising the steps of a) applying an amount of a fluid sample comprising a first member of an affinity binding pair and having a volume of about 1 to about lOOμL to the entry port of the rotatable microsystem platform and allowing the metering capillary and overflow capillary to fill by capillary action; b) rotating the platform at a first rotation speed for a time sufficient to displace the fluid in the entry port and the overflow capillary into the overflow chamber; c) rotating the platform at a second rotational speed that is greater than the first rotational speed displace a volume of the fluid in the metering capillary into the incubation chamber, wherein the incubation chamber contains a second member of the affinity binding pair; d) incubating the fluid in the incubation chamber for a time sufficient to form the affinity binding pair; e) rotating the platform at a third rotational speed that is greater than the second rotational speed to displace a volume of the wash fluid into the incubation chamber; f) rotating the platform at a fourth rotational speed that is greater than the third rotational speed to displace a volume of the reagent into the incubation chamber; g) incubating the reagent in the incubation chamber for a time sufficient to produce an amount of a detectable product that is proportional to the amount of affinity binding; and h) detecting affinity binding in the incubation chamber.
81. A resistive heater element comprising in combination: a). an electrically inert substrate capable of being screen printed with a conductive ink and a resistive ink; b) a conductive ink screen-printed in a pattern; c) a resistive ink screen-printed in a pattern over the conductive ink pattern wherein the resistive ink in electrical contact with the conductive ink and wherein an electrical potential applied across the conductive ink causes cuπent to flow across the resistive ink wherein the resistive ink produces heat.
82. The resistive heater element according to Claimδl wherein the conductive ink is a silver conductive ink selected from the group consisting of Dupont 5028, Dupont 5025, Acheson 423SS, Acheson 426SS and Acheson SS24890.
83. The resistive heater element according to Claim 81 wherein the resistive ink is selected from the group consisting of Dupont 7082, Dupont 7102, Dupont 7271, Dupont 7278 and Dupont 7285
84. A resistive heater element according to Claim 81 wherein the resistive ink is a PTC ink
85. A resistive heater element according to Claim 81 further comprising d) a dielectric ink screen-printed over the resistive ink pattern and conductive ink pattern.
86. A heat-activated wax valve in a microfluidics aπay of a microsystems platform comprising, in combination a) an amount of a solid, semi-solid or viscous liquid hydrocarbon positioned to occlude the lumen of a channel or capillary; b) a wax recrystallization chamber positioned in a channel or capillary in fluid contact therein wherein the recrystallization chamber is positioned radially more distant from the center of the platform than the wax valve; c) a resistive heater element screen-printed and in thermal contact with the wax valve and at least a portion o the wax recrystallization chamber, wherein the resistive heater element is in electrical contact with a voltage source; wherein heating the resistive heater element by applying a voltage to the element produced sufficient heat to melt the wax valve, and wherein rotation of the platform motivates the melted wax valve into the wax recrystallization chamber, and wherein the resistive heater element heats the channel or capillary and a portion of the recrystallization chamber, whereby the melted wax valve does not recrystallize in the channel or capillary and thereby does not occlude the lumen of the channel or capillary.
87. A electronic spindle for a centrifugal rotor comprising a first electrically non-conductive plate having a first side operatively attached to a mechanical spindle and having a second side opposite thereto and having embedded therein a multiplicity of electrically conductive posts having a first end that is embedded in the electrically non-conductive plate an alternating aπay of electrically conductive and non-conductive plates having the conductive posts passing there through, wherein each of the conductive plates is in electrical contact with one of the conductive posts, and wherein none of the conductive posts is in electrical contact with more than one conductive plate, and whereby the electrically conductive plates are insulated from one another, an electrically conductive plate having the conductive posts passing there through at an end of the alternating aπay opposite to the first electrically non-conductive plate, wherein the electronic spindle is mechanically and operatively attached to a motor comprising a centrifugation apparatus, and each of the electrically conductive posts is in electrical contact with an electrical contact on a centripetally motivated microsystems platform, and each of the electrically conductive plates is in electrical contact with an electrically conductive brush that transfers an electrical signal from the brush to the electrically conductive plate and to one of the electrically conductive posts, and thereby transfers the electrical signal to a rotating microsystems platform.
88. A microsystem platform according to Claim 50, wherein the inlet and outlet capillaries are connected with the mixing chamber so that their positions in the mixing chamber are offset from one another, wherein fluid flow from the inlet capillary impinges on a wall of the mixing chamber at a position other than the position occupied by the outlet capillary, and wherein fluid flow from the outlet capillary impinges on a wall of the mixing chamber at a position other than the position occupied by the inlet capillary, thereby producing turbulent flow within the mixing chamber that mixes the fluids.
89. A microsystem platform according to Claim 54, wherein the capillaries are connected with the mixing chamber so that their positions in the mixing chamber are offset from one another, wherein fluid flow from the inlet capillary impinges on a wall of the mixing chamber at a position other than the position occupied by the outlet capillary, thereby producing turbulent flow within the mixing chamber that mixes the fluids.
90. A microsystem platform according to Claim 60, wherein the inlet and outlet capillaries are connected with the mixing chamber so that their positions in the mixing chamber are offset from one another, wherein fluid flow from the inlet capillary impinges on a wall of the mixing chamber at a position other than the position occupied by the outlet capillary, and wherein fluid flow from the outlet capillary impinges on a wall of the mixing chamber at a position other than the position occupied by the inlet capillary, thereby producing turbulent flow within the mixing chamber that mixes the fluids.
91. A microsystem platform according to Claim 64, wherein the capillaries are connected with the mixing chamber so that their positions in the mixing chamber are offset from one another, wherein fluid flow from the inlet capillary impinges on a wall of the mixing chamber at a position other than the position occupied by the outlet capillary, thereby producing turbulent flow within the mixing chamber that mixes the fluids.
PCT/US1998/010520 1997-05-23 1998-05-22 Devices and methods for using centripetal acceleration to drive fluid movement in a microfluidics system WO1998053311A2 (en)

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Cited By (96)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000069560A1 (en) * 1999-05-14 2000-11-23 Gamera Bioscience Corporation A centripetally-motivated microfluidics system for performing in vitro hybridization and amplification of nucleic acids
WO2000079285A2 (en) * 1999-06-18 2000-12-28 Gamera Bioscience Corporation Devices and methods for the performance of miniaturized homogeneous assays
WO2000078455A1 (en) * 1999-06-22 2000-12-28 Tecan Trading Ag Devices and methods for the performance of miniaturized in vitro amplification assays
WO2002041995A1 (en) * 2000-11-27 2002-05-30 Pyrosequencing Ab Microfluidic flow control
WO2002000347A3 (en) * 2000-06-28 2002-10-10 3M Innovative Properties Co Sample processing devices, systems and methods
US6503359B2 (en) 1999-03-05 2003-01-07 Burstein Technologies, Inc. Monomolecular adhesion methods for manufacturing microfabricated multilaminate devices
WO2003009107A2 (en) * 2001-07-20 2003-01-30 Burstein Technologies, Inc. Optical analysis disc and related drive assembly for performing interactive centrifugation
WO2003018198A1 (en) 2001-08-28 2003-03-06 Gyros Ab Retaining microfluidic microcavity and other microfluidic structures
JP2003518250A (en) * 1999-12-23 2003-06-03 ユィロス・アクチボラグ Micro analyzer
JP2004028589A (en) * 2002-06-21 2004-01-29 Hitachi Ltd Analytical chip and analytical device
US6706519B1 (en) 1999-06-22 2004-03-16 Tecan Trading Ag Devices and methods for the performance of miniaturized in vitro amplification assays
US6734401B2 (en) 2000-06-28 2004-05-11 3M Innovative Properties Company Enhanced sample processing devices, systems and methods
EP1468286A2 (en) * 2001-12-18 2004-10-20 Serenex, Inc. Integrated protein affinity capture centrifugation device
US6888951B1 (en) 1999-08-23 2005-05-03 Nagaoka & Co., Ltd. Methods and apparatus for analyzing operational and analyte data acquired from optical disc
US6919058B2 (en) 2001-08-28 2005-07-19 Gyros Ab Retaining microfluidic microcavity and other microfluidic structures
WO2005065827A1 (en) * 2004-01-06 2005-07-21 Gyros Patent Ab Contact heating arrangement
US6965433B2 (en) 2000-11-16 2005-11-15 Nagaoka & Co., Ltd. Optical biodiscs with reflective layers
US6985672B2 (en) 2000-11-23 2006-01-10 Gyros Ab Device and method for the controlled heating in micro channel systems
US6990290B2 (en) 2000-11-23 2006-01-24 Gyros Ab Device for thermal cycling
US7005265B1 (en) 2002-06-20 2006-02-28 Wenhong Fan Nonenzymatic catalytic signal amplification for nucleic acid hybridization assays
US7033747B2 (en) 2001-04-11 2006-04-25 Nagaoka & Co., Ltd Multi-parameter assays including analysis discs and methods relating thereto
US7054258B2 (en) 2000-12-08 2006-05-30 Nagaoka & Co., Ltd. Optical disc assemblies for performing assays
US7079468B2 (en) 2000-12-08 2006-07-18 Burstein Technologies, Inc. Optical discs for measuring analytes
WO2006075966A1 (en) 2005-01-17 2006-07-20 Gyros Patent Ab A versatile flow path
US7091034B2 (en) 2000-12-15 2006-08-15 Burstein Technologies, Inc. Detection system for disk-based laboratory and improved optical bio-disc including same
WO2006110093A1 (en) * 2005-04-14 2006-10-19 Gyros Patent Ab Liquid plugs
WO2006110095A1 (en) 2005-04-14 2006-10-19 Gyros Patent Ab A microfluidic device with finger valves
US7141416B2 (en) 2001-07-12 2006-11-28 Burstein Technologies, Inc. Multi-purpose optical analysis optical bio-disc for conducting assays and various reporting agents for use therewith
US7189368B2 (en) 2001-09-17 2007-03-13 Gyros Patent Ab Functional unit enabling controlled flow in a microfluidic device
US7332326B1 (en) 1999-05-14 2008-02-19 Tecan Trading Ag Centripetally-motivated microfluidics system for performing in vitro hybridization and amplification of nucleic acids
US7369471B2 (en) 2005-05-09 2008-05-06 Searete Llc Method and system for fluid mediated disk activation and deactivation
US7390464B2 (en) 2003-06-19 2008-06-24 Burstein Technologies, Inc. Fluidic circuits for sample preparation including bio-discs and methods relating thereto
EP1946830A1 (en) * 2005-11-07 2008-07-23 Konica Minolta Medical & Graphic, Inc. Microreactor and method of liquid feeding making use of the same
US7429354B2 (en) 2001-03-19 2008-09-30 Gyros Patent Ab Structural units that define fluidic functions
US7431889B2 (en) 2003-01-30 2008-10-07 Gyros Patent Ab Inner walls of microfluidic devices
EP2019320A1 (en) 2007-07-27 2009-01-28 Hewlett-Packard Development Company, L.P. Systems and methods for measuring glycated hemoglobin
US7708874B2 (en) 2006-01-13 2010-05-04 Panasonic Corporation Electrophoresis apparatus
US7749442B2 (en) 2001-12-14 2010-07-06 Arkray, Inc. Sample measuring device
US7754151B2 (en) 2005-04-04 2010-07-13 Panasonic Corporation Liquid homogenizer and analyzer employing the same
US7754474B2 (en) 2005-07-05 2010-07-13 3M Innovative Properties Company Sample processing device compression systems and methods
US7763210B2 (en) 2005-07-05 2010-07-27 3M Innovative Properties Company Compliant microfluidic sample processing disks
US7767937B2 (en) 2005-07-05 2010-08-03 3M Innovative Properties Company Modular sample processing kits and modules
EP2237037A1 (en) 2005-12-12 2010-10-06 Gyros Patent Ab Microfluidic device and use thereof
EP2261663A2 (en) 2003-03-23 2010-12-15 Gyros Patent Ab Method for quantifying a plurality of different analytes
US7939018B2 (en) 2000-06-28 2011-05-10 3M Innovative Properties Company Multi-format sample processing devices and systems
US8003051B2 (en) 2001-12-28 2011-08-23 3M Innovative Properties Company Thermal structure for sample processing systems
US8057759B2 (en) 2006-09-05 2011-11-15 Samsung Electronics Co., Ltd. Microfluidic system and apparatus and method of controlling the same
US8133438B2 (en) 2004-01-29 2012-03-13 Gyros Patent Ab Flow paths comprising one or two porous beds
US8168136B2 (en) 2004-11-29 2012-05-01 Canon Kabushiki Kaisha Fluid-conveying apparatus and method for conveying fluid by using the same
WO2012131556A1 (en) * 2011-03-24 2012-10-04 Biosurfit, S.A. Control of liquid flow sequence on microfluidic device
WO2012158990A1 (en) * 2011-05-18 2012-11-22 3M Innovative Properties Company Systems and methods for volumetric metering on a sample processing device
WO2012164552A1 (en) 2011-06-03 2012-12-06 Radisens Diagnostics Ltd. Microfluidic disc for use in with bead-based immunoassays
US8414848B2 (en) 2007-05-10 2013-04-09 Panasonic Corporation Substrate including channel part having chamber, and multistage liquid feed device comprising the same
CN103158353A (en) * 2011-12-09 2013-06-19 施乐公司 Printhead having particle circulation with separation
US8486350B2 (en) 2008-05-09 2013-07-16 Konica Minolta Medical & Graphic, Inc. Microchip, microchip liquid supply system, and microchip liquid supply method
US8580575B2 (en) 2007-04-13 2013-11-12 Shimadzu Corporation Reactor plate and reaction processing method
US8592219B2 (en) 2005-01-17 2013-11-26 Gyros Patent Ab Protecting agent
CN103499702A (en) * 2007-11-08 2014-01-08 松下电器产业株式会社 Analyzing device
CN101520454B (en) * 2008-02-04 2014-02-26 深圳微点生物技术有限公司 Centrifugal fluid analyzer rotor
WO2014086951A1 (en) 2012-12-05 2014-06-12 Radisens Diagnostics Limited Reference and normalisation method for use with bead-based immunoassays in a microfluidic disc
US20140199783A1 (en) * 2001-03-19 2014-07-17 Gyros Patent Ab Characterization of reaction variables
ITMI20131541A1 (en) * 2013-09-19 2015-03-20 Eugenio Iannone DIAGNOSIS DEVICE, PARTICULARLY OF THE LAB-ON-CHIP TYPE.
US9040288B2 (en) 2006-03-24 2015-05-26 Handylab, Inc. Integrated system for processing microfluidic samples, and method of using the same
US9067205B2 (en) 2011-05-18 2015-06-30 3M Innovative Properties Company Systems and methods for valving on a sample processing device
US9080207B2 (en) 2006-03-24 2015-07-14 Handylab, Inc. Microfluidic system for amplifying and detecting polynucleotides in parallel
WO2015170753A1 (en) * 2014-05-08 2015-11-12 国立大学法人大阪大学 Heat convection-generating chip and liquid-weighing instrument
WO2015197618A1 (en) * 2014-06-27 2015-12-30 Hemocue Ab Device and method for determination of an analyte in blood
US9308530B2 (en) 2007-03-02 2016-04-12 Shimadzu Corporation Reaction container plate and reaction treatment apparatus
US9550184B2 (en) 2007-02-05 2017-01-24 Shimadzu Corporation Reactor plate and reaction processing method
US9618139B2 (en) 2007-07-13 2017-04-11 Handylab, Inc. Integrated heater and magnetic separator
US9670528B2 (en) 2003-07-31 2017-06-06 Handylab, Inc. Processing particle-containing samples
US9701957B2 (en) 2007-07-13 2017-07-11 Handylab, Inc. Reagent holder, and kits containing same
US9765389B2 (en) 2011-04-15 2017-09-19 Becton, Dickinson And Company Scanning real-time microfluidic thermocycler and methods for synchronized thermocycling and scanning optical detection
US9802199B2 (en) 2006-03-24 2017-10-31 Handylab, Inc. Fluorescence detector for microfluidic diagnostic system
US9815057B2 (en) 2006-11-14 2017-11-14 Handylab, Inc. Microfluidic cartridge and method of making same
US9933348B2 (en) 2011-12-08 2018-04-03 Biosurfit, S.A. Sequential aliqoting and determination of an indicator of sedimentation rate
US10052630B2 (en) 2003-03-23 2018-08-21 Gyros Patent Ab Preloaded microfluidic devices
US10065185B2 (en) 2007-07-13 2018-09-04 Handylab, Inc. Microfluidic cartridge
US10071376B2 (en) 2007-07-13 2018-09-11 Handylab, Inc. Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples
US10076754B2 (en) 2011-09-30 2018-09-18 Becton, Dickinson And Company Unitized reagent strip
USD831843S1 (en) 2011-09-30 2018-10-23 Becton, Dickinson And Company Single piece reagent holder
US10179910B2 (en) 2007-07-13 2019-01-15 Handylab, Inc. Rack for sample tubes and reagent holders
US10234474B2 (en) 2007-07-13 2019-03-19 Handylab, Inc. Automated pipetting apparatus having a combined liquid pump and pipette head system
US10351901B2 (en) 2001-03-28 2019-07-16 Handylab, Inc. Systems and methods for thermal actuation of microfluidic devices
US10364456B2 (en) 2004-05-03 2019-07-30 Handylab, Inc. Method for processing polynucleotide-containing samples
US10571935B2 (en) 2001-03-28 2020-02-25 Handylab, Inc. Methods and systems for control of general purpose microfluidic devices
CN111372685A (en) * 2018-08-30 2020-07-03 株式会社Lg化学 High-speed screening analysis system for reaction optimization
JP2020522704A (en) * 2017-11-20 2020-07-30 エルジー・ケム・リミテッド Device and method for qualitative and quantitative analysis of heavy metals utilizing rotary disk system
US10822644B2 (en) 2012-02-03 2020-11-03 Becton, Dickinson And Company External files for distribution of molecular diagnostic tests and determination of compatibility between tests
US10900066B2 (en) 2006-03-24 2021-01-26 Handylab, Inc. Microfluidic system for amplifying and detecting polynucleotides in parallel
US10935149B2 (en) 2018-03-15 2021-03-02 University Of Washington Temperature-actuated valve, fluidic device, and related methods of use
WO2021152333A1 (en) * 2020-01-31 2021-08-05 ODx Innovations Limited Centrifugally motivated fluidic systems, devices and methods
EP3895801A1 (en) * 2020-04-18 2021-10-20 CapitalBio Corporation Valve system for driving fluid and method for using the same
EP3825808A4 (en) * 2018-07-20 2022-04-20 Daegu Gyeongbuk Institute of Science and Technology Centrifugal valve control device
US11453906B2 (en) 2011-11-04 2022-09-27 Handylab, Inc. Multiplexed diagnostic detection apparatus and methods
US11806718B2 (en) 2006-03-24 2023-11-07 Handylab, Inc. Fluorescence detector for microfluidic diagnostic system

Families Citing this family (370)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5960344A (en) * 1993-12-20 1999-09-28 Norand Corporation Local area network having multiple channel wireless access
US6048734A (en) * 1995-09-15 2000-04-11 The Regents Of The University Of Michigan Thermal microvalves in a fluid flow method
US6429007B1 (en) * 1997-05-02 2002-08-06 BIOMéRIEUX, INC. Nucleic acid amplification reaction station for disposable test devices
US6632399B1 (en) * 1998-05-22 2003-10-14 Tecan Trading Ag Devices and methods for using centripetal acceleration to drive fluid movement in a microfluidics system for performing biological fluid assays
JP3469585B2 (en) * 1997-05-23 2003-11-25 ガメラ バイオサイエンス コーポレイション Apparatus and method for using centripetal acceleration to drive flow motion in microfluidics systems
US7396508B1 (en) * 2000-07-12 2008-07-08 Ventana Medical Systems, Inc. Automated molecular pathology apparatus having independent slide heaters
US6572830B1 (en) 1998-10-09 2003-06-03 Motorola, Inc. Integrated multilayered microfludic devices and methods for making the same
DE60034033T2 (en) * 1999-04-06 2007-12-06 University of Alabama, Birmingham Research Foundation, Birmingham DEVICE FOR SCREENING CRYSTALLIZATION CONDITIONS IN SOLUTIONS FOR CRYSTAL BREEDING
SE516321C2 (en) * 1999-05-31 2001-12-17 Gambro Inc Centrifuge for the treatment of blood and blood components
CA2378091A1 (en) * 1999-07-16 2001-01-25 Timothy M. Woudenberg High density electrophoresis device and method
US7088650B1 (en) 1999-08-23 2006-08-08 Worthington Mark O Methods and apparatus for optical disc data acquisition using physical synchronization markers
US6875619B2 (en) * 1999-11-12 2005-04-05 Motorola, Inc. Microfluidic devices comprising biochannels
US6361958B1 (en) * 1999-11-12 2002-03-26 Motorola, Inc. Biochannel assay for hybridization with biomaterial
WO2001055713A2 (en) * 2000-01-26 2001-08-02 Transgenomic, Inc. Method and apparatus for separating polynucleotides using monolithic capillary columns
US20020050456A1 (en) * 2000-05-15 2002-05-02 Sheppard Norman F. Use of vapor-deposited conformal coatings in microfluidic structures
US6818435B2 (en) * 2000-05-15 2004-11-16 Tecan Trading Ag Microfluidics devices and methods for performing cell based assays
US20020151078A1 (en) * 2000-05-15 2002-10-17 Kellogg Gregory J. Microfluidics devices and methods for high throughput screening
US6627159B1 (en) * 2000-06-28 2003-09-30 3M Innovative Properties Company Centrifugal filling of sample processing devices
EP1301268A2 (en) * 2000-07-03 2003-04-16 Xeotron Corporation Devices and methods for carrying out chemical reactions using photogenerated reagents
US8262989B2 (en) * 2000-07-21 2012-09-11 Senzime Ab (Publ.) Micro-calorimeter apparatus
FR2812306B1 (en) * 2000-07-28 2005-01-14 Gabriel Festoc POLYMERSIS CHAIN AMPLIFICATION SYSTEM OF TARGET NUCLEIC SEQUENCES
US6561479B1 (en) * 2000-08-23 2003-05-13 Micron Technology, Inc. Small scale actuators and methods for their formation and use
US6939451B2 (en) * 2000-09-19 2005-09-06 Aclara Biosciences, Inc. Microfluidic chip having integrated electrodes
US6855553B1 (en) 2000-10-02 2005-02-15 3M Innovative Properties Company Sample processing apparatus, methods and systems
US6796733B2 (en) 2000-10-31 2004-09-28 International Imaging Materials Inc. Thermal transfer ribbon with frosting ink layer
US6990904B2 (en) * 2000-10-31 2006-01-31 International Imaging Materials, Inc Thermal transfer assembly for ceramic imaging
US6854386B2 (en) * 2000-10-31 2005-02-15 International Imaging Materials Inc. Ceramic decal assembly
US8097471B2 (en) * 2000-11-10 2012-01-17 3M Innovative Properties Company Sample processing devices
AU2002241602A1 (en) * 2000-11-16 2002-06-11 Burstein Technologies, Inc. Methods and apparatus for detecting and quantifying lymphocytes with optical biodiscs
US7087203B2 (en) * 2000-11-17 2006-08-08 Nagaoka & Co., Ltd. Methods and apparatus for blood typing with optical bio-disc
US7026131B2 (en) * 2000-11-17 2006-04-11 Nagaoka & Co., Ltd. Methods and apparatus for blood typing with optical bio-discs
US6521188B1 (en) * 2000-11-22 2003-02-18 Industrial Technology Research Institute Microfluidic actuator
WO2002042780A2 (en) * 2000-11-22 2002-05-30 Burstein Technologies, Inc. Apparatus and methods for separating agglutinants and disperse particles
US20020076354A1 (en) * 2000-12-01 2002-06-20 Cohen David Samuel Apparatus and methods for separating components of particulate suspension
AU2002231189A1 (en) * 2000-12-22 2002-07-08 Burstein Technologies, Inc. Optical bio-discs and methods relating thereto
US6770434B2 (en) 2000-12-29 2004-08-03 The Provost, Fellows And Scholars Of The College Of The Holy & Undivided Trinity Of Queen Elizabeth Near Dublin Biological assay method
CA2433350A1 (en) * 2001-01-02 2002-08-01 President And Fellows Of Harvard College Method and apparatus for measurement of the sulfate concentration in air samples
US6653625B2 (en) * 2001-03-19 2003-11-25 Gyros Ab Microfluidic system (MS)
US20040099310A1 (en) * 2001-01-05 2004-05-27 Per Andersson Microfluidic device
US6692700B2 (en) 2001-02-14 2004-02-17 Handylab, Inc. Heat-reduction methods and systems related to microfluidic devices
JP4199544B2 (en) * 2001-03-19 2008-12-17 ユィロス・パテント・アクチボラグ Microfluidic system (EDI)
US6717136B2 (en) 2001-03-19 2004-04-06 Gyros Ab Microfludic system (EDI)
US7010391B2 (en) 2001-03-28 2006-03-07 Handylab, Inc. Methods and systems for control of microfluidic devices
US7323140B2 (en) 2001-03-28 2008-01-29 Handylab, Inc. Moving microdroplets in a microfluidic device
US6852287B2 (en) 2001-09-12 2005-02-08 Handylab, Inc. Microfluidic devices having a reduced number of input and output connections
US6806088B2 (en) * 2001-04-09 2004-10-19 Matsushita Electric Industrial Co., Ltd. Method and apparatus for improving the performance of microanalytic and microsynthetic procedures
US20030118804A1 (en) * 2001-05-02 2003-06-26 3M Innovative Properties Company Sample processing device with resealable process chamber
US6805841B2 (en) 2001-05-09 2004-10-19 The Provost Fellows And Scholars Of The College Of The Holy And Undivided Trinity Of Queen Elizabeth Near Dublin Liquid pumping system
DE10123259A1 (en) * 2001-05-12 2002-11-21 Eppendorf Ag Microfluidic storage and / or dosing component
US7060292B2 (en) * 2001-06-09 2006-06-13 Gesynthesis, Inc. Lipid structures and uses thereof
WO2003027723A2 (en) * 2001-07-24 2003-04-03 Burstein Technologies, Inc. Method and apparatus for bonded fluidic circuit for optical bio-disc
US6825127B2 (en) 2001-07-24 2004-11-30 Zarlink Semiconductor Inc. Micro-fluidic devices
SE0104077D0 (en) * 2001-10-21 2001-12-05 Gyros Ab A method and instrumentation for micro dispensation of droplets
US20030129665A1 (en) * 2001-08-30 2003-07-10 Selvan Gowri Pyapali Methods for qualitative and quantitative analysis of cells and related optical bio-disc systems
US20030143637A1 (en) * 2001-08-31 2003-07-31 Selvan Gowri Pyapali Capture layer assemblies for cellular assays including related optical analysis discs and methods
EP1423699A4 (en) * 2001-09-07 2006-01-18 Burstein Technologies Inc Nuclear morphology based identification and quantitation of white blood cell types using optical bio-disc systems
US7338760B2 (en) * 2001-10-26 2008-03-04 Ntu Ventures Private Limited Sample preparation integrated chip
EP2287330A1 (en) * 2001-11-01 2011-02-23 Rensselaer Polytechnic Institute Biocatalytic Solgel Microarrays
US20030175947A1 (en) * 2001-11-05 2003-09-18 Liu Robin Hui Enhanced mixing in microfluidic devices
EP1448992A4 (en) * 2001-11-20 2005-11-02 Burstein Technologies Inc Optical bio-discs and fluidic circuits for analysis of cells and methods relating thereto
GB0128350D0 (en) 2001-11-27 2002-01-16 Lab901 Ltd Non-rigid apparatus for microfluidic applications
US7244393B2 (en) 2001-12-21 2007-07-17 Kimberly-Clark Worldwide, Inc. Diagnostic device and system
JP3952019B2 (en) * 2001-12-28 2007-08-01 株式会社日立ハイテクノロジーズ Extraction apparatus, chemical analysis apparatus, and chemical analysis method
US6532997B1 (en) 2001-12-28 2003-03-18 3M Innovative Properties Company Sample processing device with integral electrophoresis channels
EP1461414B1 (en) * 2001-12-31 2009-04-22 The Provost Fellows and Scholars of the College of the Holy Undivided Trinity of Queen Elizabeth Near Dublin Assembly for cell-based assays
US20050221048A1 (en) * 2002-01-31 2005-10-06 James Rodney Norton Manufacturing processes for making optical analysis discs including successive patterning operations and optical discs thereby manufactured
US20050176059A1 (en) * 2002-01-31 2005-08-11 Pal Andrew A. Bio-safe dispenser and optical analysis disc assembly
WO2003065355A2 (en) * 2002-01-31 2003-08-07 Burstein Technologies, Inc. Bio-safety features for optical analysis disc and disc system including same
US20050170490A1 (en) * 2002-01-31 2005-08-04 Chen Yihfar Processes for manufacturing optical analysis discs with molded microfluidic structures and discs made according thereto
US20040241381A1 (en) * 2002-01-31 2004-12-02 Chen Yihfar Microfluidic structures with circumferential grooves for bonding adhesives and related optical analysis discs
WO2003068857A1 (en) * 2002-02-15 2003-08-21 Bridgestone Corporation Rubber composition and pneumatic tire made therefrom
JP3933058B2 (en) * 2002-02-25 2007-06-20 日立化成工業株式会社 Support unit for microfluidic system and method for manufacturing the same
US7459127B2 (en) * 2002-02-26 2008-12-02 Siemens Healthcare Diagnostics Inc. Method and apparatus for precise transfer and manipulation of fluids by centrifugal and/or capillary forces
US7419821B2 (en) 2002-03-05 2008-09-02 I-Stat Corporation Apparatus and methods for analyte measurement and immunoassay
ITVE20020012A1 (en) * 2002-03-13 2003-09-15 Alper Srl ELECTRIC DEANANT OR INSECTICIDE EMANATOR.
US20030180965A1 (en) * 2002-03-25 2003-09-25 Levent Yobas Micro-fluidic device and method of manufacturing and using the same
CA2480990A1 (en) * 2002-04-11 2003-10-23 Sequenom, Inc. Methods and devices for performing chemical reactions on a solid support
CN1650173B (en) * 2002-04-30 2012-06-06 爱科来株式会社 Analysis instrument, sample analysis method and analysis device using the instrument, and method of forming opening in the instrument
CN1551982A (en) * 2002-05-08 2004-12-01 株式会社日立高新技术 Chemical analysis equipment and gene diagnosis equipment
US7384602B2 (en) 2002-05-08 2008-06-10 Hitachi High-Technologies Corporation Chemical analysis apparatus and genetic diagnostic apparatus
JP4095886B2 (en) * 2002-05-08 2008-06-04 株式会社日立ハイテクノロジーズ Chemical analysis device and genetic diagnosis device
US20050106714A1 (en) * 2002-06-05 2005-05-19 Zarur Andrey J. Rotatable reactor systems and methods
US20040011650A1 (en) * 2002-07-22 2004-01-22 Frederic Zenhausern Method and apparatus for manipulating polarizable analytes via dielectrophoresis
CN1242848C (en) * 2002-07-22 2006-02-22 松下电器产业株式会社 Component separating device, method of manufacturing the device, and method of separating fine solid component by using the device
US7198759B2 (en) * 2002-07-26 2007-04-03 Applera Corporation Microfluidic devices, methods, and systems
US7201881B2 (en) * 2002-07-26 2007-04-10 Applera Corporation Actuator for deformable valves in a microfluidic device, and method
US7135147B2 (en) * 2002-07-26 2006-11-14 Applera Corporation Closing blade for deformable valve in a microfluidic device and method
AU2003259289A1 (en) * 2002-07-30 2004-02-16 University Of Washington Apparatus and methods for binding molecules and cells
EP1391237B1 (en) * 2002-08-01 2011-09-21 Tosoh Corporation Fine channel device, desksize chemical plant and fine particle producing apparatus employing them
TW590982B (en) * 2002-09-27 2004-06-11 Agnitio Science & Technology I Micro-fluid driving device
KR100444751B1 (en) * 2002-11-11 2004-08-16 한국전자통신연구원 Device of Controlling Fluid using Surface Tension
CN101158695A (en) 2002-12-04 2008-04-09 斯宾克斯公司 Devices and methods for programmable microscale manipulation of fluids
US7285255B2 (en) * 2002-12-10 2007-10-23 Ecolab Inc. Deodorizing and sanitizing employing a wicking device
US7125711B2 (en) * 2002-12-19 2006-10-24 Bayer Healthcare Llc Method and apparatus for splitting of specimens into multiple channels of a microfluidic device
US7507376B2 (en) * 2002-12-19 2009-03-24 3M Innovative Properties Company Integrated sample processing devices
US7094354B2 (en) * 2002-12-19 2006-08-22 Bayer Healthcare Llc Method and apparatus for separation of particles in a microfluidic device
US7476361B2 (en) * 2002-12-24 2009-01-13 Tecan Trading Ag Microfluidics devices and methods of diluting samples and reagents
US7332129B2 (en) * 2003-01-09 2008-02-19 3M Innovative Properties Company Sample processing device having process chambers with bypass slots
DE10302721A1 (en) * 2003-01-23 2004-08-05 Steag Microparts Gmbh Microfluidic arrangement for dosing liquids
KR100959101B1 (en) * 2003-02-20 2010-05-25 삼성전자주식회사 Polymerase chain reaction device and method for regulating opening or shutting of inlet and outlet of PCR device
JP2007501407A (en) * 2003-03-03 2007-01-25 長岡実業株式会社 Methods and apparatus used for the detection and quantification of various cell types and the use of optical biodiscs to do this
WO2004085668A2 (en) * 2003-03-20 2004-10-07 Northeastern Ohio Universities College Of Medecine Self-contained assay device for rapid detection of biohazardous agents
JP3888632B2 (en) * 2003-03-26 2007-03-07 靖浩 堀池 Micromixer, sample analysis kit and manufacturing method thereof
US20050037484A1 (en) * 2003-04-23 2005-02-17 Norbert Staimer Optical bio-discs including spiral fluidic circuits for performing assays
US8309039B2 (en) * 2003-05-14 2012-11-13 James Russell Webster Valve structure for consistent valve operation of a miniaturized fluid delivery and analysis system
US7435381B2 (en) * 2003-05-29 2008-10-14 Siemens Healthcare Diagnostics Inc. Packaging of microfluidic devices
US7736889B2 (en) * 2003-06-10 2010-06-15 The United States Of America As Represented By The Secretary Of The Navy Fluidic force discrimination
JP4151483B2 (en) * 2003-06-10 2008-09-17 ソニー株式会社 Bioassay substrate and bioassay apparatus and method
FR2856046B1 (en) * 2003-06-16 2005-07-29 Biomerieux Sa FLUIDIC MICROVANNE WITH OPENING BY ELECTRICAL CONTROL
EP1656203A2 (en) * 2003-06-19 2006-05-17 Nagaoka & Co., Ltd. Fluidic circuits for sample preparation including bio-discs and methods relating thereto
US20040265171A1 (en) * 2003-06-27 2004-12-30 Pugia Michael J. Method for uniform application of fluid into a reactive reagent area
US20080257754A1 (en) * 2003-06-27 2008-10-23 Pugia Michael J Method and apparatus for entry of specimens into a microfluidic device
EP1644184A2 (en) * 2003-06-27 2006-04-12 Nagaoka & Co., Ltd. Fluidic circuits, methods and apparatus for use of whole blood samples in colorimetric assays
US20040265172A1 (en) * 2003-06-27 2004-12-30 Pugia Michael J. Method and apparatus for entry and storage of specimens into a microfluidic device
US7238269B2 (en) * 2003-07-01 2007-07-03 3M Innovative Properties Company Sample processing device with unvented channel
US7077996B2 (en) * 2003-07-15 2006-07-18 Randall Brandon L Methods and apparatus for blood separation and analysis using membranes on an optical bio-disc
US20070274863A1 (en) * 2003-07-25 2007-11-29 Horacio Kido Fluidic circuits for sample preparation including bio-discs and methods relating thereto
US20060210426A1 (en) * 2003-08-05 2006-09-21 Taiyo Yuden Co., Ltd. Sample analyzer and disk-like sample analyzing medium
US7347617B2 (en) * 2003-08-19 2008-03-25 Siemens Healthcare Diagnostics Inc. Mixing in microfluidic devices
EP1508373A3 (en) * 2003-08-19 2005-08-03 Herbener, Heinz-Gerd Sample carrier with channel for liquids
JP3959436B2 (en) * 2003-08-29 2007-08-15 独立行政法人物質・材料研究機構 Flow fluctuation structure and micromixer
JPWO2005027593A1 (en) * 2003-09-12 2007-11-15 日本碍子株式会社 Plasma generating electrode and plasma reactor
EP1673167A2 (en) * 2003-09-15 2006-06-28 Tecan Trading AG Microfluidics devices and methods for performing cell based assays
US7296592B2 (en) * 2003-09-16 2007-11-20 Eksigent Technologies, Llc Composite polymer microfluidic control device
EP1669733B1 (en) * 2003-10-03 2019-01-16 National Institute for Materials Science Measuring chip
JP4391790B2 (en) * 2003-10-03 2009-12-24 独立行政法人物質・材料研究機構 Chip usage and inspection chip
WO2005036182A1 (en) 2003-10-15 2005-04-21 Matsushita Electric Industrial Co., Ltd. Method of passing fluid in capillary chip
GB0327094D0 (en) * 2003-11-21 2003-12-24 Inverness Medical Switzerland Laminated device
US7196295B2 (en) * 2003-11-21 2007-03-27 Watlow Electric Manufacturing Company Two-wire layered heater system
US7329391B2 (en) * 2003-12-08 2008-02-12 Applera Corporation Microfluidic device and material manipulating method using same
US20050130177A1 (en) * 2003-12-12 2005-06-16 3M Innovative Properties Company Variable valve apparatus and methods
US7322254B2 (en) * 2003-12-12 2008-01-29 3M Innovative Properties Company Variable valve apparatus and methods
US7837947B2 (en) * 2003-12-12 2010-11-23 3M Innovative Properties Company Sample mixing on a microfluidic device
DE10359160A1 (en) * 2003-12-16 2005-07-21 Roche Diagnostics Gmbh Test element for the examination of sample material
US7939249B2 (en) * 2003-12-24 2011-05-10 3M Innovative Properties Company Methods for nucleic acid isolation and kits using a microfluidic device and concentration step
US20050142570A1 (en) * 2003-12-24 2005-06-30 3M Innovative Properties Company Methods for nucleic acid isolation and kits using a microfluidic device and sedimenting reagent
US20050142571A1 (en) * 2003-12-24 2005-06-30 3M Innovative Properties Company Methods for nucleic acid isolation and kits using solid phase material
JP4750412B2 (en) * 2003-12-25 2011-08-17 モレックス インコーポレイテド Biologically derived molecules, dioxins and endocrine disrupting substance detection apparatus and detection method using the apparatus
CN100571860C (en) * 2004-02-18 2009-12-23 日立化成工业株式会社 Micro fluid system support unit
JP4482926B2 (en) * 2004-02-20 2010-06-16 富士フイルム株式会社 Scientific phenomenon evaluation apparatus, diffusion velocity measurement experimental apparatus, and manufacturing method thereof
US20070095393A1 (en) * 2004-03-30 2007-05-03 Piero Zucchelli Devices and methods for programmable microscale manipulation of fluids
US7056473B2 (en) * 2004-04-29 2006-06-06 Response Biomedical Corp. Method and apparatus of quantitative assays
AU2005241080B2 (en) 2004-05-03 2011-08-11 Handylab, Inc. Processing polynucleotide-containing samples
US7491363B2 (en) * 2004-07-15 2009-02-17 Applied Biosystems, Llc. Fluid processing device
US7932090B2 (en) * 2004-08-05 2011-04-26 3M Innovative Properties Company Sample processing device positioning apparatus and methods
WO2007001378A2 (en) * 2004-09-20 2007-01-04 Boston Microfluidics Microfluidic device for detecting soluble molecules
DE102004046396A1 (en) * 2004-09-24 2006-04-13 Land Baden-Württemberg, vertreten durch das Ministerium für Wissenschaft, Forschung und Kunst Baden-Württemberg, vertreten durch den Minister A particle sedimentation apparatus and method for performing a particle sedimentation
WO2006035800A1 (en) * 2004-09-30 2006-04-06 Arkray, Inc. Thin film heater and analytical instrument
EP1824601B1 (en) * 2004-10-18 2016-12-28 Life Technologies Corporation A device including a dissolvable structure for flow control
US7361315B2 (en) * 2004-10-26 2008-04-22 Konica Minolta Medical & Graphic, Inc. Micro-reactor for biological substance inspection and biological substance inspection device
EP1812604A4 (en) * 2004-11-03 2011-02-23 Third Wave Tech Inc Single step detection assay
JP4687653B2 (en) * 2004-11-30 2011-05-25 日立化成工業株式会社 Analysis pretreatment parts
CN101664662A (en) * 2004-12-09 2010-03-10 日立化成工业株式会社 Support unit for micro fluid system and process for producing the same
JP4646204B2 (en) * 2005-01-27 2011-03-09 ブラザー工業株式会社 Receiving target, sorting device, and sorting method
WO2006080140A1 (en) 2005-01-28 2006-08-03 Matsushita Electric Industrial Co., Ltd. Blood treatment apparatus and blood supply method
KR100647320B1 (en) * 2005-02-05 2006-11-23 삼성전자주식회사 Microsystem for separating serum from blood
US20060204403A1 (en) * 2005-02-28 2006-09-14 Careside Medical, Llc Micro-fluidic fluid separation device and method
EP1899702A2 (en) * 2005-04-09 2008-03-19 Boehringer Ingelheim microParts GmbH Device and method for analyzing a sample liquid
WO2006110096A1 (en) * 2005-04-14 2006-10-19 Gyros Patent Ab Microfluidic device with serial valve
JP2008538319A (en) * 2005-04-14 2008-10-23 ギロス・パテント・エービー Separation structure
US20080286156A1 (en) * 2005-04-14 2008-11-20 Gyros Patent Ab Upward Microconduits
US7565596B2 (en) 2005-09-09 2009-07-21 Searete Llc Data recovery systems
US8220014B2 (en) 2005-05-09 2012-07-10 The Invention Science Fund I, Llc Modifiable memory devices having limited expected lifetime
US9396752B2 (en) 2005-08-05 2016-07-19 Searete Llc Memory device activation and deactivation
US7668068B2 (en) 2005-06-09 2010-02-23 Searete Llc Rotation responsive disk activation and deactivation mechanisms
US7694316B2 (en) * 2005-05-09 2010-04-06 The Invention Science Fund I, Llc Fluid mediated disk activation and deactivation mechanisms
US7916592B2 (en) * 2005-05-09 2011-03-29 The Invention Science Fund I, Llc Fluid mediated disk activation and deactivation mechanisms
US8140745B2 (en) 2005-09-09 2012-03-20 The Invention Science Fund I, Llc Data retrieval methods
US8121016B2 (en) 2005-05-09 2012-02-21 The Invention Science Fund I, Llc Rotation responsive disk activation and deactivation mechanisms
US7907486B2 (en) 2006-06-20 2011-03-15 The Invention Science Fund I, Llc Rotation responsive disk activation and deactivation mechanisms
US7916615B2 (en) 2005-06-09 2011-03-29 The Invention Science Fund I, Llc Method and system for rotational control of data storage devices
US7748012B2 (en) 2005-05-09 2010-06-29 Searete Llc Method of manufacturing a limited use data storing device
US7770028B2 (en) 2005-09-09 2010-08-03 Invention Science Fund 1, Llc Limited use data storing device
US7512959B2 (en) * 2005-05-09 2009-03-31 Searete Llc Rotation responsive disk activation and deactivation mechanisms
US8218262B2 (en) 2005-05-09 2012-07-10 The Invention Science Fund I, Llc Method of manufacturing a limited use data storing device including structured data and primary and secondary read-support information
US8159925B2 (en) 2005-08-05 2012-04-17 The Invention Science Fund I, Llc Limited use memory device with associated information
US8099608B2 (en) 2005-05-09 2012-01-17 The Invention Science Fund I, Llc Limited use data storing device
US7668069B2 (en) * 2005-05-09 2010-02-23 Searete Llc Limited use memory device with associated information
US8462605B2 (en) 2005-05-09 2013-06-11 The Invention Science Fund I, Llc Method of manufacturing a limited use data storing device
US20060263265A1 (en) * 2005-05-23 2006-11-23 Der-Ren Kang Blood micro-separator
EP1899064A2 (en) * 2005-06-03 2008-03-19 Spinx, Inc. Dosimeter for programmable microscale manipulation of fluids
US7935318B2 (en) * 2005-06-13 2011-05-03 Hewlett-Packard Development Company, L.P. Microfluidic centrifugation systems
US20060281192A1 (en) * 2005-06-13 2006-12-14 Harding Philip H Method for mixing fluids in microfluidic systems
US7437914B2 (en) * 2005-06-28 2008-10-21 Hewlett-Packard Development Company, L.P. Microfluidic test systems with gas bubble reduction
CN101213447B (en) * 2005-06-28 2012-09-26 三星电子股份有限公司 Blo drive apparatus, and assay method using the same
JP4546889B2 (en) * 2005-07-08 2010-09-22 ローム株式会社 Chip with weighing unit
JP4619224B2 (en) * 2005-07-27 2011-01-26 パナソニック株式会社 Rotational analysis device
US20070031293A1 (en) * 2005-08-04 2007-02-08 Beatty Christopher C Method and apparatus for collecting and diluting a liquid sample
US20070031282A1 (en) * 2005-08-04 2007-02-08 Piero Zucchelli Devices and methods for interfacing microfluidic devices with fluid handling devices
US7731910B2 (en) * 2005-08-05 2010-06-08 Hewlett-Packard Development Company, L.P. Microfluidic mixing assembly
US7791728B2 (en) * 2005-08-11 2010-09-07 Hewlett-Packard Development Company, L.P. System for optically analyzing a substance with a selected single-wavelength
JP4802925B2 (en) * 2005-08-19 2011-10-26 パナソニック株式会社 Analytical device and analytical apparatus using the same
JP4973800B2 (en) * 2005-08-19 2012-07-11 パナソニック株式会社 Analytical device and analytical apparatus using the same
JP2007098223A (en) * 2005-09-30 2007-04-19 Fujifilm Corp Fluid operation method of chemical device
US8123834B2 (en) 2005-10-06 2012-02-28 The Board Of Trustees Of The University Of Illinois High gain selective metal organic framework preconcentrators
JP4531677B2 (en) * 2005-10-24 2010-08-25 パナソニック株式会社 Assay method, sensor device for implementing the same, and measuring device for sensor device
JP4942662B2 (en) * 2005-10-28 2012-05-30 アークレイ株式会社 Liquid feeding method and cartridge used therefor
JPWO2007052647A1 (en) * 2005-11-01 2009-04-30 パナソニック株式会社 Sample solution analysis disk and mixed sample solution analysis method
JPWO2007052648A1 (en) * 2005-11-02 2009-04-30 パナソニック株式会社 Sample analysis disc
JP4331158B2 (en) * 2005-11-15 2009-09-16 シャープ株式会社 Blade cleaning jig
US20070113908A1 (en) * 2005-11-18 2007-05-24 The Ohio State University And Bioloc, Inc. Valve for microfluidic chips
US8741136B2 (en) * 2006-02-10 2014-06-03 Boehringer Ingelheim Microparts Gmbh Device and method for treating or cleaning sample material, in particular nucleic acids
TWI272464B (en) * 2006-03-01 2007-02-01 Instr Technology Res Ct Nat Ap Fluid order controlling apparatus and method
US7651869B2 (en) * 2006-03-14 2010-01-26 Research International, Inc. Optical assay apparatus and methods
US8088616B2 (en) 2006-03-24 2012-01-03 Handylab, Inc. Heater unit for microfluidic diagnostic system
US20070224591A1 (en) * 2006-03-27 2007-09-27 General Electric Company Rotation-based microsampler, system and method of using the same
US7998433B2 (en) * 2006-04-04 2011-08-16 Samsung Electronics Co., Ltd. Valve unit and apparatus having the same
KR100763922B1 (en) * 2006-04-04 2007-10-05 삼성전자주식회사 Valve unit and apparatus with the same
JP4754394B2 (en) 2006-04-14 2011-08-24 ローム株式会社 Microchip
KR100738113B1 (en) * 2006-05-10 2007-07-12 삼성전자주식회사 Phase transition type valve and the manufacturing method thereof
US8137626B2 (en) * 2006-05-19 2012-03-20 California Institute Of Technology Fluorescence detector, filter device and related methods
US8432777B2 (en) 2006-06-19 2013-04-30 The Invention Science Fund I, Llc Method and system for fluid mediated disk activation and deactivation
US8264928B2 (en) 2006-06-19 2012-09-11 The Invention Science Fund I, Llc Method and system for fluid mediated disk activation and deactivation
KR101335920B1 (en) * 2006-08-02 2013-12-03 삼성전자주식회사 Thin film chemical analysis apparatus and analysis method using the same
EP1884284A1 (en) * 2006-08-04 2008-02-06 Samsung Electronics Co., Ltd. Closing valve unit and reaction apparatus having closing valve
US8464760B2 (en) 2006-08-16 2013-06-18 Samsung Electronic Co., Ltd. Valve unit, reaction apparatus with the same, and method of forming valve in channel
US7951333B2 (en) * 2006-09-05 2011-05-31 Samsung Electronics Co., Ltd. Centrifugal force-based microfluidic device for protein detection and microfluidic system including the same
KR101343034B1 (en) * 2006-09-05 2013-12-18 삼성전자 주식회사 Centrifugal microfluidic device for target protein detection and microfluidic system comprising the same
US8273310B2 (en) * 2006-09-05 2012-09-25 Samsung Electronics Co., Ltd. Centrifugal force-based microfluidic device for nucleic acid extraction and microfluidic system including the microfluidic device
KR100851980B1 (en) * 2006-09-05 2008-08-12 삼성전자주식회사 Centrifugal force based microfluidic device having heat-activative unit, microfluidic system comprinsing the same and method for driving the system
WO2008036614A1 (en) * 2006-09-18 2008-03-27 California Institute Of Technology Apparatus for detecting target molecules and related methods
US7857141B2 (en) 2006-12-11 2010-12-28 Samsung Electronics Co., Ltd. Apparatus and method for separating components
US8307988B2 (en) * 2006-12-11 2012-11-13 Samsung Electronics Co., Ltd. Apparatus and method for separating components
KR100818290B1 (en) * 2006-12-11 2008-03-31 삼성전자주식회사 Component separating apparatus and method
TW200844420A (en) 2006-12-22 2008-11-16 3M Innovative Properties Co Enhanced sample processing devices, systems and methods
EP2117713B1 (en) 2006-12-22 2019-08-07 DiaSorin S.p.A. Thermal transfer methods for microfluidic systems
US20080241844A1 (en) * 2007-02-06 2008-10-02 Network Biosystems, Inc. Devices and Methods for the Performance of Miniaturized In Vitro Assays
KR101305976B1 (en) * 2007-02-12 2013-09-12 삼성전자주식회사 Centrifugal force based microfluidic device for serial dilution and microfluidic system comprising the same
WO2008121446A2 (en) * 2007-02-14 2008-10-09 The Board Of Trustees Of The University Of Illinois Portable electrochemical multiphase microreactor for sensing trace chemical vapors
US7799656B2 (en) 2007-03-15 2010-09-21 Dalsa Semiconductor Inc. Microchannels for BioMEMS devices
KR100883658B1 (en) 2007-04-02 2009-02-18 삼성전자주식회사 Centrifugal force-based microfluidic device and microfluidic system including the same
ES2748134T3 (en) 2007-04-04 2020-03-13 Ande Corp Systems for rapid multiple amplification of target nucleic acids
US8168444B2 (en) * 2007-04-09 2012-05-01 Panasonic Corporation Substrate having channel portion including chambers, and method of transferring liquid by using the substrate
WO2008132873A1 (en) * 2007-04-13 2008-11-06 Shimadzu Corporation Reaction container plate and reaction processing method
JP4947140B2 (en) * 2007-04-13 2012-06-06 株式会社島津製作所 Reaction vessel plate and reaction processing method
KR100858091B1 (en) * 2007-04-24 2008-09-10 삼성전자주식회사 Centrifugal force-based microfluidic device having sample distribution structure and microfluidic system including the microfluidic device
EP2140001A2 (en) * 2007-04-25 2010-01-06 3M Innovative Properties Company Methods for nucleic acid amplification
KR101258434B1 (en) 2007-05-03 2013-05-02 삼성전자주식회사 Microfluidic system and fabricating method of the same
US8124030B2 (en) * 2007-05-08 2012-02-28 The Regents Of The University Of California Microfluidic device having regulated fluid transfer between elements located therein
US7980272B2 (en) * 2007-06-21 2011-07-19 Samsung Electronics Co., Ltd. Microfluidic valve, method of manufacturing the same, and microfluidic device comprising the microfluidic valve
US7829867B2 (en) * 2007-07-02 2010-11-09 Caridianbct Biotechnologies, Llc Apparatus for photo reduction of contaminants in blood and blood products with calibration means
JPWO2009008128A1 (en) * 2007-07-06 2010-09-02 パナソニック株式会社 Liquid feeding device and liquid feeding method
US20090136385A1 (en) 2007-07-13 2009-05-28 Handylab, Inc. Reagent Tube
US8133671B2 (en) 2007-07-13 2012-03-13 Handylab, Inc. Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples
USD621060S1 (en) 2008-07-14 2010-08-03 Handylab, Inc. Microfluidic cartridge
EP3141901B1 (en) 2007-10-04 2018-12-12 PHC Holdings Corporation Analysis device, and analysis apparatus and method using the same
JP5013423B2 (en) * 2007-10-18 2012-08-29 ローム株式会社 Microchip
US8381169B2 (en) * 2007-10-30 2013-02-19 International Business Machines Corporation Extending unified process and method content to include dynamic and collaborative content
TWI362491B (en) * 2007-11-02 2012-04-21 Ind Tech Res Inst Fluid analytical device and fluid analytical method thereof
SE531873C2 (en) * 2007-11-12 2009-09-01 Lifeassays Ab Device for biochemical processing and analysis of sample liquid
EP2214801A4 (en) * 2007-11-22 2016-12-21 Samsung Electronics Co Ltd Thin film valve device and its controlling apparatus
CN101450336B (en) * 2007-12-06 2012-02-08 财团法人工业技术研究院 Fluid separation method and device
EP2072131B1 (en) * 2007-12-13 2015-04-22 Roche Diagnostics GmbH Microfluid element for mixing a fluid into a reagent
JP4872922B2 (en) * 2008-01-07 2012-02-08 株式会社島津製作所 Reaction vessel
JP4872921B2 (en) * 2008-01-07 2012-02-08 株式会社島津製作所 Reaction vessel
JP4872923B2 (en) * 2008-01-07 2012-02-08 株式会社島津製作所 Reaction vessel
DE102008003979B3 (en) * 2008-01-11 2009-06-10 Hahn-Schickard-Gesellschaft für angewandte Forschung e.V. Fluidic device, fluidic module and method for handling a fluid
US8123841B2 (en) 2008-01-16 2012-02-28 The Board Of Trustees Of The University Of Illinois Column design for micro gas chromatograph
WO2009093838A2 (en) * 2008-01-21 2009-07-30 Thin-film layered centrifuge device and analysis method using the same
US8632243B2 (en) * 2008-03-10 2014-01-21 The Hong Kong Polytechnic University Microfluidic mixing using continuous acceleration/deceleration methodology
US8735846B1 (en) 2008-03-14 2014-05-27 Advanced Technology Applications, Llc Electromagnetic biosensor
US7812318B1 (en) 2008-03-14 2010-10-12 Advanced Technology Applications, Llc Electromagnetic biosensor
US7854893B2 (en) * 2008-03-28 2010-12-21 Panasonic Corporation Analysis device and an analysis apparatus using the analysis device
KR100919400B1 (en) * 2008-04-07 2009-09-29 삼성전자주식회사 Microfluidic device and method for fabricating the same
US8269029B2 (en) 2008-04-08 2012-09-18 The Board Of Trustees Of The University Of Illinois Water repellent metal-organic frameworks, process for making and uses regarding same
KR100960066B1 (en) * 2008-05-14 2010-05-31 삼성전자주식회사 Microfluidic device containing lyophilized reagent therein and analysing method using the same
KR100955481B1 (en) 2008-05-14 2010-04-30 삼성전자주식회사 Valve unit, microfluidic device with the same, and method for fabricating the valve unit
TWI385383B (en) 2008-05-28 2013-02-11 Ind Tech Res Inst Analytical system, and analytical method and flowing structure thereof
JP5239552B2 (en) * 2008-06-26 2013-07-17 株式会社島津製作所 Reaction vessel plate and reaction processing method
USD618820S1 (en) 2008-07-11 2010-06-29 Handylab, Inc. Reagent holder
KR20100007809A (en) * 2008-07-14 2010-01-22 삼성전자주식회사 Microfluidic device, sample analyzing method using the same, and dilution ratio measuring method
USD787087S1 (en) 2008-07-14 2017-05-16 Handylab, Inc. Housing
JP5430569B2 (en) * 2008-07-29 2014-03-05 シャープ株式会社 Microdevice and microchip apparatus, and analysis method using them
US7947492B2 (en) * 2008-08-20 2011-05-24 Northeastern Ohio Universities College Of Medicine Device improving the detection of a ligand
KR101390717B1 (en) * 2008-09-02 2014-04-30 삼성전자주식회사 Microfluidic device and method of loading sample thereto
US9289765B2 (en) * 2008-09-23 2016-03-22 Samsung Electronics Co., Ltd. Micro-fluidic device and sample testing apparatus using the same
WO2010042355A1 (en) * 2008-10-06 2010-04-15 Response Biomedical Corporation Sample preparation device
US20100088855A1 (en) * 2008-10-14 2010-04-15 Magna Mirrors Of America, Inc. Vehicle door handle assembly
GB2464721C (en) 2008-10-23 2013-08-14 Biosurfit Sa Jet deflection device
GB2466644B (en) 2008-12-30 2011-05-11 Biosurfit Sa Liquid handling
US7927904B2 (en) 2009-01-05 2011-04-19 Dalsa Semiconductor Inc. Method of making BIOMEMS devices
KR101578149B1 (en) * 2009-01-29 2015-12-17 삼성전자주식회사 Microfluidic valve unit for controlling fluid flow and method for fabricating the same
KR20100102781A (en) * 2009-03-12 2010-09-27 삼성전자주식회사 Centrifugal force based microfluidic device, and method for fabricating the same
WO2010132167A1 (en) * 2009-05-11 2010-11-18 Caridianbct Biotechnologies, Llc Stable calibration means for apparatus for photo reduction of contaminants in blood
US20100297745A1 (en) * 2009-05-19 2010-11-25 Nanyang Technological University Flow-through method of functionalizing inner surfaces of a microfluidic device
US9550985B2 (en) 2009-06-15 2017-01-24 Netbio, Inc. Methods for forensic DNA quantitation
TWI360438B (en) * 2009-08-25 2012-03-21 Ind Tech Res Inst Analytical system, analytical method and flow-path
JP5311406B2 (en) * 2009-09-10 2013-10-09 独立行政法人産業技術総合研究所 Immunosensor
KR20110043034A (en) * 2009-10-20 2011-04-27 삼성전자주식회사 Microfluidic device and method of testing sample using the same
US20120282682A1 (en) * 2009-11-02 2012-11-08 Ffei Limited Micro-channel structure method and apparatus
US20110117607A1 (en) * 2009-11-13 2011-05-19 3M Innovative Properties Company Annular compression systems and methods for sample processing devices
USD667561S1 (en) 2009-11-13 2012-09-18 3M Innovative Properties Company Sample processing disk cover
USD638951S1 (en) 2009-11-13 2011-05-31 3M Innovative Properties Company Sample processing disk cover
US8834792B2 (en) 2009-11-13 2014-09-16 3M Innovative Properties Company Systems for processing sample processing devices
USD638550S1 (en) 2009-11-13 2011-05-24 3M Innovative Properties Company Sample processing disk cover
KR101422573B1 (en) * 2009-11-26 2014-07-25 삼성전자 주식회사 Centrifugal Micro-fluidic Device and Method to measure biological makers from liquid specimen
GB2476474B (en) 2009-12-22 2012-03-28 Biosurfit Sa Surface plasmon resonance detection system
US20110292963A1 (en) * 2010-01-28 2011-12-01 Conductive Compounds, Inc. Laser position detection system
JP2011155921A (en) * 2010-02-02 2011-08-18 Seiko Epson Corp Biochip, specimen reactor, and method for specimen reaction
GB201003614D0 (en) * 2010-03-04 2010-04-21 Airbus Operations Ltd Water drain tool
GB2479139A (en) 2010-03-29 2011-10-05 Biosurfit Sa A liquid distribution and metering device
JP6049621B2 (en) * 2010-09-26 2016-12-21 ダ・ユー・エンタープライジズ、エルエルシー Analyte separation
KR20120091631A (en) * 2011-02-09 2012-08-20 삼성전자주식회사 Microfluidic device
KR20120093636A (en) * 2011-02-15 2012-08-23 삼성전자주식회사 Microfluidic device
EP3270141B1 (en) * 2011-03-08 2020-05-20 Universite Laval Fluidic centripetal device
CA2830103C (en) * 2011-03-15 2020-03-24 National Research Council Of Canada Microfluidic system having monolithic nanoplasmonic structures
EP2688675A1 (en) * 2011-03-24 2014-01-29 Boehringer Ingelheim Microparts GmbH Device and method for removing a supernatant of a liquid sample, and use of a valve device having a soluble membrane
JP5740650B2 (en) * 2011-05-17 2015-06-24 国立研究開発法人産業技術総合研究所 Disk type microfluidic chip and measurement system using the same
MX337943B (en) 2011-05-18 2016-03-29 Focus Diagnostics Inc Systems and methods for detecting the presence of a selected volume of material in a sample processing device.
USD672467S1 (en) 2011-05-18 2012-12-11 3M Innovative Properties Company Rotatable sample processing disk
GB2491813A (en) 2011-06-03 2012-12-19 Univ Dublin City A microfluidic device with sacrificial valve
JP2013033687A (en) 2011-08-03 2013-02-14 Sharp Corp Storage battery module
US9822890B2 (en) 2011-08-30 2017-11-21 The Royal Institution For The Advancement Of Learning/Mcgill University Method and system for pre-programmed self-power microfluidic circuits
US10384209B2 (en) * 2011-09-15 2019-08-20 The Chinese University Of Hong Kong Microfluidic platform and method for controlling the same
JP5465758B2 (en) * 2011-09-26 2014-04-09 富士フイルム株式会社 Thyroxine immunoassay using fluorescent particles
WO2013053953A1 (en) * 2011-10-14 2013-04-18 Université de Liège Method for measuring beta-lactam antibiotics
JP5951219B2 (en) * 2011-10-24 2016-07-13 ローム株式会社 Microchip with built-in liquid reagent
EP2786120B1 (en) * 2011-12-01 2019-02-13 Biosurfit, S.A. Photometric device and method
DE102012202775B4 (en) * 2012-02-23 2016-08-25 Hahn-Schickard-Gesellschaft für angewandte Forschung e.V. FLUIDIKMODUL, DEVICE AND METHOD FOR PUMPING A LIQUID
RU2015107730A (en) 2012-08-08 2016-09-27 Конинклейке Филипс Н.В. CENTRIFUGAL MICROFLUIDIC DEVICE AND METHODS OF APPLICATION
DK3425401T3 (en) 2012-10-08 2023-10-23 Bl Technologies Inc PREFILLED TEST SUBSTRATES FOR TESTING LAL-REACTIVE SUBSTANCES, USING METHODS AND MANUFACTURING METHODS
KR20140055528A (en) * 2012-10-31 2014-05-09 삼성전자주식회사 Microfluidic structure, microfluidic system and control method for microfluidic test device
US10888862B2 (en) 2012-12-05 2021-01-12 Radisens Diagnostics Limited Acceleration-primed valving system for centrifugal microfluidics
US10130947B2 (en) * 2012-12-05 2018-11-20 Radisens Diagnostics Limited Valving system for use in centrifugal microfluidic platforms
KR101441594B1 (en) * 2012-12-27 2014-09-23 매쓰파워 주식회사 Heat sensitive polymer composite based Nucleic Acid Amplification disc apparatus and analysis method using the same
US20160290962A1 (en) * 2013-03-21 2016-10-06 Nec Corporation Method and apparatus for electrophoresis
DE102013206622B4 (en) * 2013-04-13 2017-10-19 Pr Germany Gmbh Closure element in the form of a window or a door
JP6244589B2 (en) * 2013-05-21 2017-12-13 国立大学法人名古屋大学 Micro-channel chip for separating fine particles, advection integrated unit, system for separating fine particles, and method for separating fine particles
US9308508B2 (en) 2013-07-22 2016-04-12 Kianoosh Peyvan Sequential delivery device and method
KR20150027939A (en) * 2013-09-04 2015-03-13 삼성전자주식회사 Microfluidic apparatus
US10076751B2 (en) * 2013-12-30 2018-09-18 General Electric Company Systems and methods for reagent storage
GB201401422D0 (en) * 2014-01-28 2014-03-12 Secr Defence Method and device for heating a chemical reaction
AU2015221851B2 (en) * 2014-02-25 2019-05-09 C.Y. O'connor Erade Village Foundation Inc. Methods and systems for measuring melting temperatures
EP3163306A4 (en) 2014-06-30 2018-01-24 Panasonic Healthcare Holdings Co., Ltd. Substrate for sample analysis, and sample analysis apparatus
US10539582B2 (en) * 2014-06-30 2020-01-21 Phc Holdings Corporation Substrate for sample analysis, sample analysis device, sample analysis system, and method for removing liquid from liquid that contains magnetic particles
US10309976B2 (en) 2014-06-30 2019-06-04 Phc Holdings Corporation Substrate for sample analysis, sample analysis device, sample analysis system, and program for sample analysis system
JP6588910B2 (en) 2014-06-30 2019-10-09 Phcホールディングス株式会社 Sample analysis substrate, sample analysis apparatus, sample analysis system, and program for sample analysis system
CN105717313A (en) * 2014-12-04 2016-06-29 广明光电股份有限公司 Biological disc detection drive device and method
CN107209193B (en) 2014-12-12 2019-08-02 普和希控股公司 Sample analysis substrate, sample analyzer, sample analysis system and sample analysis system program
US9736888B2 (en) * 2015-03-12 2017-08-15 The Boeing Company Composite panel with integrated heater and associated methods for manufacturing
US10356847B2 (en) 2015-03-12 2019-07-16 The Boeing Company Composite panel with integrated heater system and associated methods for manufacturing
DK3311160T3 (en) * 2015-06-20 2021-03-15 Capitainer Ab A PLASMA SEPARATOR MICROFLUID DEVICE
CA2991470A1 (en) * 2015-08-07 2017-02-16 Poc Medical Systems, Inc. Microfluidic devices and methods of use thereof
US10501715B1 (en) * 2015-09-11 2019-12-10 Mark H. Widick System for the formation of fibrin foam
USD799715S1 (en) 2015-10-23 2017-10-10 Gene POC, Inc. Fluidic centripetal device
WO2017127248A1 (en) * 2016-01-18 2017-07-27 The Board Of Trustees Of The Leland Stanford Junior University Paperfuge: an integrated paper-based centrifugation and microfluidics platform for low-cost diagnostics
CN109477821A (en) 2016-03-18 2019-03-15 奎多心血管股份有限公司 Microfluidic device, system and method
US10368396B2 (en) 2016-04-01 2019-07-30 The Boeing Company Heat pipe with printed heater and associated methods for manufacturing
CN107398307B (en) 2016-05-18 2023-07-25 博奥生物集团有限公司 Integrated micro-fluidic chip
CN106513063B (en) * 2016-06-06 2018-09-07 苏州汶颢芯片科技有限公司 The centrifugal chip and its mixed method of consecutive reaction can be achieved
WO2018005464A1 (en) * 2016-06-27 2018-01-04 Abaxis, Inc. Devices with modified conduits
JP6635897B2 (en) * 2016-08-30 2020-01-29 シスメックス株式会社 Sample analysis cartridge, method for producing the same, and use thereof
JP6917451B2 (en) * 2016-10-03 2021-08-11 テルモ ビーシーティー、インコーポレーテッド Fluid centrifuge
US10639635B2 (en) * 2016-10-07 2020-05-05 Biosurfit, SA Device and method for handling liquid
US10591078B2 (en) 2016-12-15 2020-03-17 The Boeing Company Fluid flow control device
US20200238279A1 (en) * 2017-03-08 2020-07-30 Northwestern University Devices, systems, and methods for specimen preparation and analysis using capillary and centrifugal forces
DE102017204002B4 (en) 2017-03-10 2019-05-23 Hahn-Schickard-Gesellschaft für angewandte Forschung e.V. CENTRIFUGO-PNEUMATIC SWITCHING OF LIQUID
WO2019017927A1 (en) * 2017-07-19 2019-01-24 Hewlett-Packard Development Company, L.P. Microfluidic fluid flow in a target fluid
JP6876328B2 (en) * 2017-08-28 2021-05-26 国立大学法人大阪大学 Container and its manufacturing method and detection method of test substance
JP6950955B2 (en) * 2017-12-28 2021-10-13 国立研究開発法人産業技術総合研究所 Assay device
JP6846388B2 (en) * 2018-06-26 2021-03-24 シスメックス株式会社 Measurement cartridge and liquid feeding method
GB201820617D0 (en) * 2018-12-18 2019-01-30 Greindx As System for capturing cells
EP3742174A4 (en) * 2019-04-04 2022-01-12 Biobank Inc. Multi-system for simultaneously performing biochemical test and blood test, and multi-disc used therefor
CN110762238B (en) * 2019-11-19 2021-05-04 华北水利水电大学 Automatic fluid flow rate control device
CA3098079C (en) * 2019-12-13 2023-06-20 Autonomous Medical Devices Inc. Apparatus and method for point-of-care, rapid, field-deployable diagnostic testing of covid-19, viruses, antibodies and markers
US20220099622A1 (en) * 2020-09-30 2022-03-31 Icare Diagnostics International Co. Ltd. Nucleic acid detection kit and nucleic acid detection device
KR20220135997A (en) * 2021-03-31 2022-10-07 주식회사 씨티셀즈 Apparatus to control cell separation
DE102021208891B3 (en) 2021-08-13 2022-10-13 Hahn-Schickard-Gesellschaft für angewandte Forschung e.V. Fluid vacuum switching
GB202118919D0 (en) * 2021-12-23 2022-02-09 Osler Diagnostics Ltd Liquid handling device
US20230285964A1 (en) * 2022-03-10 2023-09-14 Hewlett-Packard Development Company L.P. Metered volume microfluidic devices

Citations (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR1292407A (en) * 1961-03-24 1962-05-04 Rotary electric distributor
US3713062A (en) * 1970-12-29 1973-01-23 Texas Instruments Inc Snap disc thermal sequencer
GB1307628A (en) * 1970-08-21 1973-02-21 Ver Volkseigener Betriebe Elek Slip ring assembly for electrical machines
US3952116A (en) * 1974-09-17 1976-04-20 Texas Instruments Incorporated Process for forming electrical resistance heaters
US4030834A (en) * 1976-04-08 1977-06-21 The United States Of America As Represented By The United States Energy Research And Development Administration Dynamic multistation photometer
US4258740A (en) * 1977-05-16 1981-03-31 Packard Instrument Company, Inc. Fluid flow control device
US4722853A (en) * 1985-08-12 1988-02-02 Raychem Corporation Method of printing a polymer thick film ink
US4872821A (en) * 1987-03-23 1989-10-10 Gold Medal Products Co. Cotton candy machine
EP0408207A2 (en) * 1989-07-13 1991-01-16 Illinois Tool Works Inc. Positive temperature coefficient heater
US5160702A (en) * 1989-01-17 1992-11-03 Molecular Devices Corporation Analyzer with improved rotor structure
US5171533A (en) * 1989-07-31 1992-12-15 Fine Richard A Biological assay cassette and method for making same
WO1994026414A1 (en) * 1993-05-17 1994-11-24 Syntex (U.S.A.) Inc. Reaction container for specific binding assays and method for its use
WO1996007919A1 (en) * 1994-09-02 1996-03-14 Biometric Imaging, Inc. A disposable cartridge and method for an assay of a biological sample
WO1997008556A1 (en) * 1995-08-31 1997-03-06 First Medical, Inc. Analytical rotor and method for detecting analytes in liquid samples
WO1997021090A1 (en) * 1995-12-05 1997-06-12 Gamera Bioscience Devices and methods for using centripetal acceleration to drive fluid movement in a microfluidics system with on-board informatics
WO1998007019A1 (en) * 1996-08-12 1998-02-19 Gamera Bioscience Corporation Capillary microvalve

Family Cites Families (55)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3528449A (en) * 1968-02-27 1970-09-15 Trw Inc Fluid flow control apparatus
US3679367A (en) 1970-09-14 1972-07-25 Technicon Instr Apparatus for determining the pack volume of particulates in liquid mixtures
US3916152A (en) * 1972-05-31 1975-10-28 Union Carbide Corp Temperature control system for a centrifugal-type chemistry analyzer
FR2400700A1 (en) 1977-08-18 1979-03-16 Guigan Jean CONDITIONING DEVICE FOR A LIQUID SAMPLE FOR ANALYSIS
FR2409514A1 (en) 1977-11-17 1979-06-15 Bretaudiere Jean Pierre DEVELOPMENT OF CENTRIFUGATION ANALYSIS EQUIPMENT
US4381291A (en) 1979-02-23 1983-04-26 Ab Fortia Measurement of free ligands
DE3044385A1 (en) 1980-11-25 1982-06-24 Boehringer Mannheim Gmbh, 6800 Mannheim METHOD FOR CARRYING OUT ANALYTICAL PROVISIONS AND ROTOR INSERT ELEMENT SUITABLE FOR THIS
US4729862A (en) 1981-04-24 1988-03-08 Allied-Signal Inc. Nylon composition for use in rotational molding
US5496520A (en) 1982-01-08 1996-03-05 Kelton; Arden A. Rotary fluid manipulator
DE3314961A1 (en) 1983-04-25 1984-10-25 Boehringer Mannheim Gmbh, 6800 Mannheim ANALYZER FOR PHOTOMETRICALLY DETERMINING A PARAMETER OF A LIQUID
GB8317124D0 (en) 1983-06-23 1983-07-27 Ekins R P Free ligand assay
US4612959A (en) * 1985-05-07 1986-09-23 Mobil Oil Corporation Valveless shut-off and transfer device
WO1988001058A1 (en) 1986-08-06 1988-02-11 Roger Philip Ekins Determination of analyte concentration using two labelling markers
US4935346A (en) 1986-08-13 1990-06-19 Lifescan, Inc. Minimum procedure system for the determination of analytes
US4940527A (en) 1987-06-01 1990-07-10 Abbott Laboratories Two-part test cartridge for centrifuge
US5173262A (en) 1987-07-17 1992-12-22 Martin Marietta Energy Systems, Inc. Rotor assembly and method for automatically processing liquids
US5242803A (en) 1987-07-17 1993-09-07 Martin Marietta Energy Systems, Inc. Rotor assembly and assay method
CA1333850C (en) 1987-08-27 1995-01-10 Michael M. Gorin Apparatus and method for dilution and mixing of liquid samples
US4999304A (en) 1987-12-28 1991-03-12 Miles Inc. Dynamic braking centrifuge
US5252294A (en) 1988-06-01 1993-10-12 Messerschmitt-Bolkow-Blohm Gmbh Micromechanical structure
KR920700403A (en) 1989-03-07 1992-02-19 혼고오 므쯔미 Analysis device for liquid samples and analysis method for liquid samples using this analysis device
US5006749A (en) 1989-10-03 1991-04-09 Regents Of The University Of California Method and apparatus for using ultrasonic energy for moving microminiature elements
US5122284A (en) 1990-06-04 1992-06-16 Abaxis, Inc. Apparatus and method for optically analyzing biological fluids
US5061381A (en) 1990-06-04 1991-10-29 Abaxis, Inc. Apparatus and method for separating cells from biological fluids
US5186844A (en) 1991-04-01 1993-02-16 Abaxis, Inc. Apparatus and method for continuous centrifugal blood cell separation
US5173193A (en) 1991-04-01 1992-12-22 Schembri Carol T Centrifugal rotor having flow partition
US5242606A (en) 1990-06-04 1993-09-07 Abaxis, Incorporated Sample metering port for analytical rotor having overflow chamber
DE69230951T2 (en) * 1991-06-11 2001-03-01 Univ Cambridge Tech Centrifuges, in particular for geotechnical test methods
US5413732A (en) 1991-08-19 1995-05-09 Abaxis, Inc. Reagent compositions for analytical testing
US5304348A (en) 1992-02-11 1994-04-19 Abaxis, Inc. Reagent container for analytical rotor
US5195967A (en) * 1992-02-18 1993-03-23 Nakao Naomi L Anticlotting device and method for use with IV catheters
AU4047493A (en) 1992-04-02 1993-11-08 Abaxis, Inc. Analytical rotor with dye mixing chamber
US5275016A (en) 1992-04-24 1994-01-04 Abaxis, Inc. Cryogenic apparatus
AU674685B2 (en) 1992-05-01 1997-01-09 Trustees Of The University Of Pennsylvania, The Fluid handling in microfabricated analytical devices
US5304487A (en) 1992-05-01 1994-04-19 Trustees Of The University Of Pennsylvania Fluid handling in mesoscale analytical devices
US5259829A (en) * 1993-01-11 1993-11-09 Vanegmond Cornelis F H Centrifugal separating apparatus
JPH06265447A (en) 1993-03-16 1994-09-22 Hitachi Ltd Trace quantity reactor and trace element measuring instrument therewith
US5478750A (en) 1993-03-31 1995-12-26 Abaxis, Inc. Methods for photometric analysis
US5432009A (en) 1993-06-17 1995-07-11 Sekisui Kagaku Kogyo Kabushiki Kaisha Film for first-aid sticking plaster
US5368704A (en) 1993-08-06 1994-11-29 Teknekron Corporation Micro-electrochemical valves and method
US5591643A (en) 1993-09-01 1997-01-07 Abaxis, Inc. Simplified inlet channels
US5409665A (en) 1993-09-01 1995-04-25 Abaxis, Inc. Simultaneous cuvette filling with means to isolate cuvettes
US5431303A (en) * 1993-09-30 1995-07-11 Miskell; David L. Two-part aerosol dispenser employing fusible plug
US5403415A (en) 1993-11-17 1995-04-04 Abaxis, Inc. Method and device for ultrasonic welding
JP3365067B2 (en) 1994-02-10 2003-01-08 ソニー株式会社 Plasma apparatus and plasma processing method using the same
US5413009A (en) 1994-03-16 1995-05-09 Fragnito; Frank Variable speed transmission assembly
DE4410224C2 (en) 1994-03-24 1996-02-29 Meinhard Prof Dr Knoll Miniaturized flow analysis system
US5590052A (en) 1994-04-14 1996-12-31 Abaxis, Inc. Error checking in blood analyzer
CA2192196C (en) 1994-06-06 2004-11-23 Anne R. Kopf-Sill Modified siphons for improved metering precision
US5639428A (en) 1994-07-19 1997-06-17 Becton Dickinson And Company Method and apparatus for fully automated nucleic acid amplification, nucleic acid assay and immunoassay
US5593378A (en) * 1995-03-07 1997-01-14 Dyck; Howard F. Centrifugal separator for flowable mixtures and having magnets and housing scrapers
US5622819A (en) * 1995-03-28 1997-04-22 Kinetic Biosystems, Inc. Centrifugal fermentation process
JP3469585B2 (en) 1997-05-23 2003-11-25 ガメラ バイオサイエンス コーポレイション Apparatus and method for using centripetal acceleration to drive flow motion in microfluidics systems
AU4713400A (en) * 1999-05-14 2000-12-05 Gamera Bioscience Corporation A centripetally-motivated microfluidics system for performing in vitro hybridization and amplification of nucleic acids
DE19944617C2 (en) * 1999-09-17 2002-01-10 Fresenius Ag Centrifuge with a magnetic gear

Patent Citations (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR1292407A (en) * 1961-03-24 1962-05-04 Rotary electric distributor
GB1307628A (en) * 1970-08-21 1973-02-21 Ver Volkseigener Betriebe Elek Slip ring assembly for electrical machines
US3713062A (en) * 1970-12-29 1973-01-23 Texas Instruments Inc Snap disc thermal sequencer
US3952116A (en) * 1974-09-17 1976-04-20 Texas Instruments Incorporated Process for forming electrical resistance heaters
US4030834A (en) * 1976-04-08 1977-06-21 The United States Of America As Represented By The United States Energy Research And Development Administration Dynamic multistation photometer
US4258740A (en) * 1977-05-16 1981-03-31 Packard Instrument Company, Inc. Fluid flow control device
US4722853A (en) * 1985-08-12 1988-02-02 Raychem Corporation Method of printing a polymer thick film ink
US4872821A (en) * 1987-03-23 1989-10-10 Gold Medal Products Co. Cotton candy machine
US5160702A (en) * 1989-01-17 1992-11-03 Molecular Devices Corporation Analyzer with improved rotor structure
EP0408207A2 (en) * 1989-07-13 1991-01-16 Illinois Tool Works Inc. Positive temperature coefficient heater
US5171533A (en) * 1989-07-31 1992-12-15 Fine Richard A Biological assay cassette and method for making same
WO1994026414A1 (en) * 1993-05-17 1994-11-24 Syntex (U.S.A.) Inc. Reaction container for specific binding assays and method for its use
WO1996007919A1 (en) * 1994-09-02 1996-03-14 Biometric Imaging, Inc. A disposable cartridge and method for an assay of a biological sample
WO1997008556A1 (en) * 1995-08-31 1997-03-06 First Medical, Inc. Analytical rotor and method for detecting analytes in liquid samples
WO1997021090A1 (en) * 1995-12-05 1997-06-12 Gamera Bioscience Devices and methods for using centripetal acceleration to drive fluid movement in a microfluidics system with on-board informatics
WO1998007019A1 (en) * 1996-08-12 1998-02-19 Gamera Bioscience Corporation Capillary microvalve

Cited By (176)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6503359B2 (en) 1999-03-05 2003-01-07 Burstein Technologies, Inc. Monomolecular adhesion methods for manufacturing microfabricated multilaminate devices
WO2000069560A1 (en) * 1999-05-14 2000-11-23 Gamera Bioscience Corporation A centripetally-motivated microfluidics system for performing in vitro hybridization and amplification of nucleic acids
US7332326B1 (en) 1999-05-14 2008-02-19 Tecan Trading Ag Centripetally-motivated microfluidics system for performing in vitro hybridization and amplification of nucleic acids
WO2000079285A2 (en) * 1999-06-18 2000-12-28 Gamera Bioscience Corporation Devices and methods for the performance of miniaturized homogeneous assays
WO2000079285A3 (en) * 1999-06-18 2002-06-20 Gamera Bioscience Corp Devices and methods for the performance of miniaturized homogeneous assays
US6582662B1 (en) 1999-06-18 2003-06-24 Tecan Trading Ag Devices and methods for the performance of miniaturized homogeneous assays
US6706519B1 (en) 1999-06-22 2004-03-16 Tecan Trading Ag Devices and methods for the performance of miniaturized in vitro amplification assays
WO2000078455A1 (en) * 1999-06-22 2000-12-28 Tecan Trading Ag Devices and methods for the performance of miniaturized in vitro amplification assays
US6888951B1 (en) 1999-08-23 2005-05-03 Nagaoka & Co., Ltd. Methods and apparatus for analyzing operational and analyte data acquired from optical disc
US7664289B2 (en) 1999-08-23 2010-02-16 Vindur Technologies, Inc. Methods and apparatus for analyzing operational and analyte data acquired from optical discs
JP2003518250A (en) * 1999-12-23 2003-06-03 ユィロス・アクチボラグ Micro analyzer
WO2002000347A3 (en) * 2000-06-28 2002-10-10 3M Innovative Properties Co Sample processing devices, systems and methods
US7939018B2 (en) 2000-06-28 2011-05-10 3M Innovative Properties Company Multi-format sample processing devices and systems
US7164107B2 (en) 2000-06-28 2007-01-16 3M Innovative Properties Company Enhanced sample processing devices, systems and methods
US6734401B2 (en) 2000-06-28 2004-05-11 3M Innovative Properties Company Enhanced sample processing devices, systems and methods
EP2316569A1 (en) * 2000-06-28 2011-05-04 3M Innovative Properties Company Enhanced sample processing devices, systems and methods
US7435933B2 (en) 2000-06-28 2008-10-14 3M Innovative Properties Company Enhanced sample processing devices, systems and methods
US6987253B2 (en) 2000-06-28 2006-01-17 3M Innovative Properties Company Enhanced sample processing devices, systems and methods
US6965433B2 (en) 2000-11-16 2005-11-15 Nagaoka & Co., Ltd. Optical biodiscs with reflective layers
US7668443B2 (en) 2000-11-23 2010-02-23 Gyros Ab Device and method for the controlled heating in micro channel systems
US6985672B2 (en) 2000-11-23 2006-01-10 Gyros Ab Device and method for the controlled heating in micro channel systems
US6990290B2 (en) 2000-11-23 2006-01-24 Gyros Ab Device for thermal cycling
WO2002041995A1 (en) * 2000-11-27 2002-05-30 Pyrosequencing Ab Microfluidic flow control
US7599275B2 (en) 2000-12-08 2009-10-06 Vindur Technologies, Inc. Optical discs for measuring analytes
US7200100B2 (en) 2000-12-08 2007-04-03 Nagaoka & Co., Ltd. Optical disc assemblies for performing assays
US7054258B2 (en) 2000-12-08 2006-05-30 Nagaoka & Co., Ltd. Optical disc assemblies for performing assays
US7079468B2 (en) 2000-12-08 2006-07-18 Burstein Technologies, Inc. Optical discs for measuring analytes
US7542383B2 (en) 2000-12-08 2009-06-02 Vindur Technologies, Inc. Optical disc assemblies for performing assays
US7889615B2 (en) 2000-12-08 2011-02-15 Vindur Technologies, Inc. Optical discs for measuring analytes
US7366063B2 (en) 2000-12-08 2008-04-29 Burstein Technologies, Inc. Optical discs for measuring analytes
US7091034B2 (en) 2000-12-15 2006-08-15 Burstein Technologies, Inc. Detection system for disk-based laboratory and improved optical bio-disc including same
US7429354B2 (en) 2001-03-19 2008-09-30 Gyros Patent Ab Structural units that define fluidic functions
US20140199783A1 (en) * 2001-03-19 2014-07-17 Gyros Patent Ab Characterization of reaction variables
US10620194B2 (en) 2001-03-19 2020-04-14 Gyros Patent Ab Characterization of reaction variables
US10619191B2 (en) 2001-03-28 2020-04-14 Handylab, Inc. Systems and methods for thermal actuation of microfluidic devices
US10351901B2 (en) 2001-03-28 2019-07-16 Handylab, Inc. Systems and methods for thermal actuation of microfluidic devices
US10571935B2 (en) 2001-03-28 2020-02-25 Handylab, Inc. Methods and systems for control of general purpose microfluidic devices
US7033747B2 (en) 2001-04-11 2006-04-25 Nagaoka & Co., Ltd Multi-parameter assays including analysis discs and methods relating thereto
US7141416B2 (en) 2001-07-12 2006-11-28 Burstein Technologies, Inc. Multi-purpose optical analysis optical bio-disc for conducting assays and various reporting agents for use therewith
WO2003009107A3 (en) * 2001-07-20 2003-11-27 Burstein Technologies Inc Optical analysis disc and related drive assembly for performing interactive centrifugation
WO2003009107A2 (en) * 2001-07-20 2003-01-30 Burstein Technologies, Inc. Optical analysis disc and related drive assembly for performing interactive centrifugation
EP2283924A1 (en) 2001-08-28 2011-02-16 Gyros Patent Ab Retaining microfluidic microcavity and other microfluidic structures
US6919058B2 (en) 2001-08-28 2005-07-19 Gyros Ab Retaining microfluidic microcavity and other microfluidic structures
EP2281633A1 (en) 2001-08-28 2011-02-09 Gyros Patent Ab Retaining microfluidic microcavity and other microfluidic structures
US7459129B2 (en) 2001-08-28 2008-12-02 Gyros Patent Ab Retaining microfluidic microcavity and other microfluidic structures
EP2269736A1 (en) 2001-08-28 2011-01-05 Gyros Patent Ab Retaining microfluidic microcavity and other microfluidic structures
WO2003018198A1 (en) 2001-08-28 2003-03-06 Gyros Ab Retaining microfluidic microcavity and other microfluidic structures
US7275858B2 (en) 2001-08-28 2007-10-02 Gyros Patent Ab Retaining microfluidic microcavity and other microfluidic structures
US7300199B2 (en) 2001-08-28 2007-11-27 Gyros Ab Retaining microfluidic microcavity and other microfluidic structures
US7189368B2 (en) 2001-09-17 2007-03-13 Gyros Patent Ab Functional unit enabling controlled flow in a microfluidic device
US7749442B2 (en) 2001-12-14 2010-07-06 Arkray, Inc. Sample measuring device
EP1468286A4 (en) * 2001-12-18 2008-08-20 Serenex Inc Integrated protein affinity capture centrifugation device
EP1468286A2 (en) * 2001-12-18 2004-10-20 Serenex, Inc. Integrated protein affinity capture centrifugation device
US8003051B2 (en) 2001-12-28 2011-08-23 3M Innovative Properties Company Thermal structure for sample processing systems
US7005265B1 (en) 2002-06-20 2006-02-28 Wenhong Fan Nonenzymatic catalytic signal amplification for nucleic acid hybridization assays
US8288156B2 (en) 2002-06-21 2012-10-16 Hitachi, Ltd. Analytical chip and analyzer
JP2004028589A (en) * 2002-06-21 2004-01-29 Hitachi Ltd Analytical chip and analytical device
US7431889B2 (en) 2003-01-30 2008-10-07 Gyros Patent Ab Inner walls of microfluidic devices
US10052630B2 (en) 2003-03-23 2018-08-21 Gyros Patent Ab Preloaded microfluidic devices
EP2261663A2 (en) 2003-03-23 2010-12-15 Gyros Patent Ab Method for quantifying a plurality of different analytes
US7390464B2 (en) 2003-06-19 2008-06-24 Burstein Technologies, Inc. Fluidic circuits for sample preparation including bio-discs and methods relating thereto
US9670528B2 (en) 2003-07-31 2017-06-06 Handylab, Inc. Processing particle-containing samples
US10865437B2 (en) 2003-07-31 2020-12-15 Handylab, Inc. Processing particle-containing samples
US10731201B2 (en) 2003-07-31 2020-08-04 Handylab, Inc. Processing particle-containing samples
US11078523B2 (en) 2003-07-31 2021-08-03 Handylab, Inc. Processing particle-containing samples
WO2005065827A1 (en) * 2004-01-06 2005-07-21 Gyros Patent Ab Contact heating arrangement
US8133438B2 (en) 2004-01-29 2012-03-13 Gyros Patent Ab Flow paths comprising one or two porous beds
US10364456B2 (en) 2004-05-03 2019-07-30 Handylab, Inc. Method for processing polynucleotide-containing samples
US10494663B1 (en) 2004-05-03 2019-12-03 Handylab, Inc. Method for processing polynucleotide-containing samples
US11441171B2 (en) 2004-05-03 2022-09-13 Handylab, Inc. Method for processing polynucleotide-containing samples
US10443088B1 (en) 2004-05-03 2019-10-15 Handylab, Inc. Method for processing polynucleotide-containing samples
US10604788B2 (en) 2004-05-03 2020-03-31 Handylab, Inc. System for processing polynucleotide-containing samples
US8168136B2 (en) 2004-11-29 2012-05-01 Canon Kabushiki Kaisha Fluid-conveying apparatus and method for conveying fluid by using the same
US8592219B2 (en) 2005-01-17 2013-11-26 Gyros Patent Ab Protecting agent
WO2006075966A1 (en) 2005-01-17 2006-07-20 Gyros Patent Ab A versatile flow path
US7754151B2 (en) 2005-04-04 2010-07-13 Panasonic Corporation Liquid homogenizer and analyzer employing the same
WO2006110093A1 (en) * 2005-04-14 2006-10-19 Gyros Patent Ab Liquid plugs
WO2006110095A1 (en) 2005-04-14 2006-10-19 Gyros Patent Ab A microfluidic device with finger valves
US8372357B2 (en) 2005-04-14 2013-02-12 Gyros Patent Ab Liquid plugs
US7369471B2 (en) 2005-05-09 2008-05-06 Searete Llc Method and system for fluid mediated disk activation and deactivation
US7763210B2 (en) 2005-07-05 2010-07-27 3M Innovative Properties Company Compliant microfluidic sample processing disks
US7754474B2 (en) 2005-07-05 2010-07-13 3M Innovative Properties Company Sample processing device compression systems and methods
US7767937B2 (en) 2005-07-05 2010-08-03 3M Innovative Properties Company Modular sample processing kits and modules
US8080409B2 (en) 2005-07-05 2011-12-20 3M Innovative Properties Company Sample processing device compression systems and methods
US7534618B2 (en) 2005-10-28 2009-05-19 Hewlett-Packard Development Company, L.P. Systems and methods for measuring glycated hemoglobin
EP1946830A4 (en) * 2005-11-07 2012-09-19 Konica Minolta Med & Graphic Microreactor and method of liquid feeding making use of the same
EP1946830A1 (en) * 2005-11-07 2008-07-23 Konica Minolta Medical & Graphic, Inc. Microreactor and method of liquid feeding making use of the same
EP2237037A1 (en) 2005-12-12 2010-10-06 Gyros Patent Ab Microfluidic device and use thereof
US7708874B2 (en) 2006-01-13 2010-05-04 Panasonic Corporation Electrophoresis apparatus
US11666903B2 (en) 2006-03-24 2023-06-06 Handylab, Inc. Integrated system for processing microfluidic samples, and method of using same
US11806718B2 (en) 2006-03-24 2023-11-07 Handylab, Inc. Fluorescence detector for microfluidic diagnostic system
US9040288B2 (en) 2006-03-24 2015-05-26 Handylab, Inc. Integrated system for processing microfluidic samples, and method of using the same
US10821436B2 (en) 2006-03-24 2020-11-03 Handylab, Inc. Integrated system for processing microfluidic samples, and method of using the same
US9080207B2 (en) 2006-03-24 2015-07-14 Handylab, Inc. Microfluidic system for amplifying and detecting polynucleotides in parallel
US10799862B2 (en) 2006-03-24 2020-10-13 Handylab, Inc. Integrated system for processing microfluidic samples, and method of using same
US10913061B2 (en) 2006-03-24 2021-02-09 Handylab, Inc. Integrated system for processing microfluidic samples, and method of using the same
US9802199B2 (en) 2006-03-24 2017-10-31 Handylab, Inc. Fluorescence detector for microfluidic diagnostic system
US10695764B2 (en) 2006-03-24 2020-06-30 Handylab, Inc. Fluorescence detector for microfluidic diagnostic system
US10843188B2 (en) 2006-03-24 2020-11-24 Handylab, Inc. Integrated system for processing microfluidic samples, and method of using the same
US11085069B2 (en) 2006-03-24 2021-08-10 Handylab, Inc. Microfluidic system for amplifying and detecting polynucleotides in parallel
US10821446B1 (en) 2006-03-24 2020-11-03 Handylab, Inc. Fluorescence detector for microfluidic diagnostic system
US11142785B2 (en) 2006-03-24 2021-10-12 Handylab, Inc. Microfluidic system for amplifying and detecting polynucleotides in parallel
US10857535B2 (en) 2006-03-24 2020-12-08 Handylab, Inc. Integrated system for processing microfluidic samples, and method of using same
US11141734B2 (en) 2006-03-24 2021-10-12 Handylab, Inc. Fluorescence detector for microfluidic diagnostic system
US10900066B2 (en) 2006-03-24 2021-01-26 Handylab, Inc. Microfluidic system for amplifying and detecting polynucleotides in parallel
US8057759B2 (en) 2006-09-05 2011-11-15 Samsung Electronics Co., Ltd. Microfluidic system and apparatus and method of controlling the same
US9815057B2 (en) 2006-11-14 2017-11-14 Handylab, Inc. Microfluidic cartridge and method of making same
US10710069B2 (en) 2006-11-14 2020-07-14 Handylab, Inc. Microfluidic valve and method of making same
US9550184B2 (en) 2007-02-05 2017-01-24 Shimadzu Corporation Reactor plate and reaction processing method
US9308530B2 (en) 2007-03-02 2016-04-12 Shimadzu Corporation Reaction container plate and reaction treatment apparatus
US8580575B2 (en) 2007-04-13 2013-11-12 Shimadzu Corporation Reactor plate and reaction processing method
US8414848B2 (en) 2007-05-10 2013-04-09 Panasonic Corporation Substrate including channel part having chamber, and multistage liquid feed device comprising the same
US9701957B2 (en) 2007-07-13 2017-07-11 Handylab, Inc. Reagent holder, and kits containing same
US10625261B2 (en) 2007-07-13 2020-04-21 Handylab, Inc. Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples
US10065185B2 (en) 2007-07-13 2018-09-04 Handylab, Inc. Microfluidic cartridge
US10071376B2 (en) 2007-07-13 2018-09-11 Handylab, Inc. Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples
US11266987B2 (en) 2007-07-13 2022-03-08 Handylab, Inc. Microfluidic cartridge
US10100302B2 (en) 2007-07-13 2018-10-16 Handylab, Inc. Polynucleotide capture materials, and methods of using same
US11549959B2 (en) 2007-07-13 2023-01-10 Handylab, Inc. Automated pipetting apparatus having a combined liquid pump and pipette head system
US10717085B2 (en) 2007-07-13 2020-07-21 Handylab, Inc. Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples
US10139012B2 (en) 2007-07-13 2018-11-27 Handylab, Inc. Integrated heater and magnetic separator
US10179910B2 (en) 2007-07-13 2019-01-15 Handylab, Inc. Rack for sample tubes and reagent holders
US11254927B2 (en) 2007-07-13 2022-02-22 Handylab, Inc. Polynucleotide capture materials, and systems using same
US10234474B2 (en) 2007-07-13 2019-03-19 Handylab, Inc. Automated pipetting apparatus having a combined liquid pump and pipette head system
US11060082B2 (en) 2007-07-13 2021-07-13 Handy Lab, Inc. Polynucleotide capture materials, and systems using same
US10844368B2 (en) 2007-07-13 2020-11-24 Handylab, Inc. Diagnostic apparatus to extract nucleic acids including a magnetic assembly and a heater assembly
US10632466B1 (en) 2007-07-13 2020-04-28 Handylab, Inc. Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples
US10625262B2 (en) 2007-07-13 2020-04-21 Handylab, Inc. Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples
US10875022B2 (en) 2007-07-13 2020-12-29 Handylab, Inc. Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples
US11466263B2 (en) 2007-07-13 2022-10-11 Handylab, Inc. Diagnostic apparatus to extract nucleic acids including a magnetic assembly and a heater assembly
US10590410B2 (en) 2007-07-13 2020-03-17 Handylab, Inc. Polynucleotide capture materials, and methods of using same
US9618139B2 (en) 2007-07-13 2017-04-11 Handylab, Inc. Integrated heater and magnetic separator
EP2019320A1 (en) 2007-07-27 2009-01-28 Hewlett-Packard Development Company, L.P. Systems and methods for measuring glycated hemoglobin
CN103499702A (en) * 2007-11-08 2014-01-08 松下电器产业株式会社 Analyzing device
US9182384B2 (en) 2007-11-08 2015-11-10 Panasonic Healthcare Holdings Co., Ltd. Analyzing device and analyzing method using same
US10101317B2 (en) 2007-11-08 2018-10-16 Phc Holdings Corporation Rotatable analyzing device with a separating cavity and a capillary cavity
CN101520454B (en) * 2008-02-04 2014-02-26 深圳微点生物技术有限公司 Centrifugal fluid analyzer rotor
US8486350B2 (en) 2008-05-09 2013-07-16 Konica Minolta Medical & Graphic, Inc. Microchip, microchip liquid supply system, and microchip liquid supply method
US9625916B2 (en) 2011-03-24 2017-04-18 Biosurfit, S.A. Control of liquid flow sequence on a microfluidic device
EP3000530A3 (en) * 2011-03-24 2016-05-11 Biosurfit, S.A. Control of liquid flow sequence on microfluidic device
CN103517762A (en) * 2011-03-24 2014-01-15 拜森菲特有限公司 Control of liquid flow sequence on microfluidic device
US9908116B2 (en) 2011-03-24 2018-03-06 Biosurfit, S.A. Control of liquid flow sequence on microfluidic device
WO2012131556A1 (en) * 2011-03-24 2012-10-04 Biosurfit, S.A. Control of liquid flow sequence on microfluidic device
US11788127B2 (en) 2011-04-15 2023-10-17 Becton, Dickinson And Company Scanning real-time microfluidic thermocycler and methods for synchronized thermocycling and scanning optical detection
US10781482B2 (en) 2011-04-15 2020-09-22 Becton, Dickinson And Company Scanning real-time microfluidic thermocycler and methods for synchronized thermocycling and scanning optical detection
US9765389B2 (en) 2011-04-15 2017-09-19 Becton, Dickinson And Company Scanning real-time microfluidic thermocycler and methods for synchronized thermocycling and scanning optical detection
WO2012158990A1 (en) * 2011-05-18 2012-11-22 3M Innovative Properties Company Systems and methods for volumetric metering on a sample processing device
US9067205B2 (en) 2011-05-18 2015-06-30 3M Innovative Properties Company Systems and methods for valving on a sample processing device
KR20190025731A (en) * 2011-05-18 2019-03-11 디아소린 에스.피.에이. Systems and methods for volumetric metering on a sample processing device
KR102110174B1 (en) 2011-05-18 2020-05-14 디아소린 에스.피.에이. Systems and methods for volumetric metering on a sample processing device
AU2012255144B2 (en) * 2011-05-18 2015-01-29 Diasorin Italia S.P.A. Systems and methods for volumetric metering on a sample processing device
WO2012164552A1 (en) 2011-06-03 2012-12-06 Radisens Diagnostics Ltd. Microfluidic disc for use in with bead-based immunoassays
US9897596B2 (en) 2011-06-03 2018-02-20 Radisens Diagnostics Limited Microfluidic disc for use in with bead-based immunoassays
US10076754B2 (en) 2011-09-30 2018-09-18 Becton, Dickinson And Company Unitized reagent strip
USD905269S1 (en) 2011-09-30 2020-12-15 Becton, Dickinson And Company Single piece reagent holder
USD831843S1 (en) 2011-09-30 2018-10-23 Becton, Dickinson And Company Single piece reagent holder
US11453906B2 (en) 2011-11-04 2022-09-27 Handylab, Inc. Multiplexed diagnostic detection apparatus and methods
US9933348B2 (en) 2011-12-08 2018-04-03 Biosurfit, S.A. Sequential aliqoting and determination of an indicator of sedimentation rate
CN103158353A (en) * 2011-12-09 2013-06-19 施乐公司 Printhead having particle circulation with separation
CN103158353B (en) * 2011-12-09 2015-12-02 施乐公司 Phase transformation ink fountain and printhead
US10822644B2 (en) 2012-02-03 2020-11-03 Becton, Dickinson And Company External files for distribution of molecular diagnostic tests and determination of compatibility between tests
WO2014086951A1 (en) 2012-12-05 2014-06-12 Radisens Diagnostics Limited Reference and normalisation method for use with bead-based immunoassays in a microfluidic disc
ITMI20131541A1 (en) * 2013-09-19 2015-03-20 Eugenio Iannone DIAGNOSIS DEVICE, PARTICULARLY OF THE LAB-ON-CHIP TYPE.
US10046321B2 (en) 2013-09-19 2018-08-14 Dianax S.R.L. Diagnostic device, particularly of the lab-on-chip type
WO2015040009A1 (en) * 2013-09-19 2015-03-26 Eugenio Iannone Diagnostic device, particularly of the lab-on-chip type
US10493416B2 (en) 2014-05-08 2019-12-03 Osaka University Thermal convection generating chip and liquid measuring device
WO2015170753A1 (en) * 2014-05-08 2015-11-12 国立大学法人大阪大学 Heat convection-generating chip and liquid-weighing instrument
JPWO2015170753A1 (en) * 2014-05-08 2017-04-20 国立大学法人大阪大学 Thermal convection generating chip and liquid weighing tool
WO2015197618A1 (en) * 2014-06-27 2015-12-30 Hemocue Ab Device and method for determination of an analyte in blood
JP2020522704A (en) * 2017-11-20 2020-07-30 エルジー・ケム・リミテッド Device and method for qualitative and quantitative analysis of heavy metals utilizing rotary disk system
US10935149B2 (en) 2018-03-15 2021-03-02 University Of Washington Temperature-actuated valve, fluidic device, and related methods of use
EP3825808A4 (en) * 2018-07-20 2022-04-20 Daegu Gyeongbuk Institute of Science and Technology Centrifugal valve control device
CN111372685A (en) * 2018-08-30 2020-07-03 株式会社Lg化学 High-speed screening analysis system for reaction optimization
WO2021152333A1 (en) * 2020-01-31 2021-08-05 ODx Innovations Limited Centrifugally motivated fluidic systems, devices and methods
EP3895801A1 (en) * 2020-04-18 2021-10-20 CapitalBio Corporation Valve system for driving fluid and method for using the same
US11565255B2 (en) 2020-04-18 2023-01-31 Capitalbio Corporation Valve system for driving fluid and method for using the same

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US6399361B2 (en) 2002-06-04
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AU7591998A (en) 1998-12-11
US6548788B2 (en) 2003-04-15
US6719682B2 (en) 2004-04-13
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US6302134B1 (en) 2001-10-16
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US6063589A (en) 2000-05-16
US20040191125A1 (en) 2004-09-30
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