US20010052976A1 - Scanning optical detection system - Google Patents

Scanning optical detection system Download PDF

Info

Publication number
US20010052976A1
US20010052976A1 US09/927,820 US92782001A US2001052976A1 US 20010052976 A1 US20010052976 A1 US 20010052976A1 US 92782001 A US92782001 A US 92782001A US 2001052976 A1 US2001052976 A1 US 2001052976A1
Authority
US
United States
Prior art keywords
microlocation
examined
radiation
diameter
excitation
Prior art date
Legal status (The legal status 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 status listed.)
Abandoned
Application number
US09/927,820
Inventor
Robert Juncosa
William Butler
Lei Wu
Robert Cormack
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nanogen Inc
Original Assignee
Nanogen Inc
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
Priority claimed from US08/146,504 external-priority patent/US5605662A/en
Priority claimed from US08/271,882 external-priority patent/US6017696A/en
Priority claimed from US08/304,657 external-priority patent/US5632957A/en
Priority claimed from US08/534,454 external-priority patent/US5849486A/en
Application filed by Nanogen Inc filed Critical Nanogen Inc
Priority to US09/927,820 priority Critical patent/US20010052976A1/en
Publication of US20010052976A1 publication Critical patent/US20010052976A1/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/6456—Spatial resolved fluorescence measurements; Imaging
    • G01N21/6458—Fluorescence microscopy
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J19/0046—Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J19/0093—Microreactors, e.g. miniaturised or microfabricated reactors
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y10/00—Nanotechnology for information processing, storage or transmission, e.g. quantum computing or single electron logic
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
    • C07K1/045—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers using devices to improve synthesis, e.g. reactors, special vessels
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
    • C07K1/047—Simultaneous synthesis of different peptide species; Peptide libraries
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6816—Hybridisation assays characterised by the detection means
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6816—Hybridisation assays characterised by the detection means
    • C12Q1/6825—Nucleic acid detection involving sensors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
    • C12Q1/6837—Enzymatic or biochemical coupling of nucleic acids to a solid phase using probe arrays or probe chips
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/6452—Individual samples arranged in a regular 2D-array, e.g. multiwell plates
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/76—Chemiluminescence; Bioluminescence
    • G—PHYSICS
    • G11—INFORMATION STORAGE
    • G11C—STATIC STORES
    • G11C13/00—Digital stores characterised by the use of storage elements not covered by groups G11C11/00, G11C23/00, or G11C25/00
    • G11C13/0002—Digital stores characterised by the use of storage elements not covered by groups G11C11/00, G11C23/00, or G11C25/00 using resistive RAM [RRAM] elements
    • G11C13/0009—RRAM elements whose operation depends upon chemical change
    • G11C13/0014—RRAM elements whose operation depends upon chemical change comprising cells based on organic memory material
    • G—PHYSICS
    • G11—INFORMATION STORAGE
    • G11C—STATIC STORES
    • G11C13/00—Digital stores characterised by the use of storage elements not covered by groups G11C11/00, G11C23/00, or G11C25/00
    • G11C13/0002—Digital stores characterised by the use of storage elements not covered by groups G11C11/00, G11C23/00, or G11C25/00 using resistive RAM [RRAM] elements
    • G11C13/0009—RRAM elements whose operation depends upon chemical change
    • G11C13/0014—RRAM elements whose operation depends upon chemical change comprising cells based on organic memory material
    • G11C13/0019—RRAM elements whose operation depends upon chemical change comprising cells based on organic memory material comprising bio-molecules
    • G—PHYSICS
    • G11—INFORMATION STORAGE
    • G11C—STATIC STORES
    • G11C19/00—Digital stores in which the information is moved stepwise, e.g. shift registers
    • H—ELECTRICITY
    • H01—ELECTRIC ELEMENTS
    • H01L—SEMICONDUCTOR DEVICES NOT COVERED BY CLASS H10
    • H01L25/00—Assemblies consisting of a plurality of individual semiconductor or other solid state devices ; Multistep manufacturing processes thereof
    • H01L25/50—Multistep manufacturing processes of assemblies consisting of devices, each device being of a type provided for in group H01L27/00 or H01L29/00
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00279—Features relating to reactor vessels
    • B01J2219/00306—Reactor vessels in a multiple arrangement
    • B01J2219/00313—Reactor vessels in a multiple arrangement the reactor vessels being formed by arrays of wells in blocks
    • B01J2219/00315—Microtiter plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00279—Features relating to reactor vessels
    • B01J2219/00306—Reactor vessels in a multiple arrangement
    • B01J2219/00313—Reactor vessels in a multiple arrangement the reactor vessels being formed by arrays of wells in blocks
    • B01J2219/00315—Microtiter plates
    • B01J2219/00317—Microwell devices, i.e. having large numbers of wells
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00497—Features relating to the solid phase supports
    • B01J2219/00527—Sheets
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00585—Parallel processes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/0059—Sequential processes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00596—Solid-phase processes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
    • B01J2219/00612—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports the surface being inorganic
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
    • B01J2219/00614—Delimitation of the attachment areas
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
    • B01J2219/00614—Delimitation of the attachment areas
    • B01J2219/00621—Delimitation of the attachment areas by physical means, e.g. trenches, raised areas
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00653—Making arrays on substantially continuous surfaces the compounds being bound to electrodes embedded in or on the solid supports
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00659—Two-dimensional arrays
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/0068—Means for controlling the apparatus of the process
    • B01J2219/00686—Automatic
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/0068—Means for controlling the apparatus of the process
    • B01J2219/00686—Automatic
    • B01J2219/00689—Automatic using computers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00709—Type of synthesis
    • B01J2219/00713—Electrochemical synthesis
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00722—Nucleotides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00725—Peptides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00731—Saccharides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/025—Align devices or objects to ensure defined positions relative to each other
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/14—Process control and prevention of errors
    • B01L2200/143—Quality control, feedback systems
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0636—Integrated biosensor, microarrays
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0645—Electrodes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0816—Cards, e.g. flat sample carriers usually with flow in two horizontal directions
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0832—Geometry, shape and general structure cylindrical, tube shaped
    • B01L2300/0838—Capillaries
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0877—Flow chambers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0415—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic
    • B01L2400/0421—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic electrophoretic flow
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/11—Compounds covalently bound to a solid support
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/06—Libraries containing nucleotides or polynucleotides, or derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/10—Libraries containing peptides or polypeptides, or derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/12—Libraries containing saccharides or polysaccharides, or derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B60/00—Apparatus specially adapted for use in combinatorial chemistry or with libraries
    • C40B60/14—Apparatus specially adapted for use in combinatorial chemistry or with libraries for creating libraries
    • G—PHYSICS
    • G11—INFORMATION STORAGE
    • G11C—STATIC STORES
    • G11C13/00—Digital stores characterised by the use of storage elements not covered by groups G11C11/00, G11C23/00, or G11C25/00
    • G11C13/04—Digital stores characterised by the use of storage elements not covered by groups G11C11/00, G11C23/00, or G11C25/00 using optical elements ; using other beam accessed elements, e.g. electron or ion beam
    • H—ELECTRICITY
    • H01—ELECTRIC ELEMENTS
    • H01L—SEMICONDUCTOR DEVICES NOT COVERED BY CLASS H10
    • H01L2924/00—Indexing scheme for arrangements or methods for connecting or disconnecting semiconductor or solid-state bodies as covered by H01L24/00
    • H01L2924/0001—Technical content checked by a classifier
    • H01L2924/0002—Not covered by any one of groups H01L24/00, H01L24/00 and H01L2224/00

Definitions

  • This invention relates to optical detection and examining systems, especially systems for examining fluorescent or chemilluminescent radiation. More particularly, the invention relates to optical systems for examining localized areas containing biological fluorescent materials, where those systems require relatively high sensitivity.
  • Molecular biology comprises a wide variety of techniques for the analysis of nucleic acid and protein. Many of these techniques and procedures form the basis of clinical diagnostic assays and tests. These techniques include nucleic acid hybridization analysis, restriction enzyme analysis, genetic sequence analysis, and the separation and purification of nucleic acids and proteins (See, e.g., J. Sambrook, E. F. Fritsch, and T. Maniatis, Molecular Cloning: A Laboratory Manual, 2 Ed., Cold spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989).
  • the complete process for carrying out a DNA hybridization analysis for a genetic or infectious disease is very involved.
  • the complete process may be divided into a number of steps and substeps, broadly including the steps of obtaining the sample, disrupting the cells within the sample, performing complexity reduction or amplification, performing some sort of assay or hybridization, followed by detection of the presence or absence of a desired event serving to generate a result.
  • an excitation energy of one wavelength is delivered to the region of interest and energy of a different wavelength is emitted and detected.
  • Large scale systems generally those having a region of interest of two millimeters or greater, have been manufactured in which the quality of the overall system is not inherently limited by the size requirements of the optical elements or the ability to place them in optical proximity to the region of interest.
  • the conventional approaches to fluorometer design have proved inadequate.
  • the excitation and emission optical elements must be placed close to the region of interest.
  • a focused spot size is relatively small, often requiring sophisticated optical designs.
  • the demands for high accuracy in mechanical alignment increase.
  • the size of the optical components required to achieve these goals in relation to their distance from the region of interest becomes important, and in many cases, compromises the performance obtained.
  • One disclosed carrier is a microtiter plate with multiple samples, e.g., 96, arranged in rows and columns.
  • Various imaging devices are disclosed, such as an imaging photon detector, microchannel plate intensifiers and charged coupled devices (CCDs).
  • the signals representing the discrete areas of reactions have the background noise signal subtracted from them.
  • Another multiple location immunoassay system is disclosed in Elings et al. U.S. Pat. No. 4,537,861 entitled “Apparatus and Method for Homogeneous Immunoassay”.
  • a spatial pattern formed by a spatial array of separate regions of antiligand material are disposed on a surface. The presence or absence of a binding reaction between a ligand and the antiligand is then detected.
  • a source of illumination is shined on the combined ligand-antiligand location, and the emitted radiation detected.
  • the contribution to the imager due to free labeled molecules plus background contaminants are suppressed through use of a chopper system in positional correlation to the examined array which generates a reference signal.
  • a scanning confocal microscope is disclosed in U.S. Pat. No. 5,296,703 entitled “Scanning Confocal Microscope Using Fluorescence Detection”.
  • a scanning confocal microscope is provided for scanning a sample with an incident beam of radiation and detecting the resulting fluorescence radiation to provide data suitable for use in a raster scanned display of the fluorescence.
  • First and second closely spaced scanning mirrors direct an incident beam to a sample and direct the fluorescent radiation towards a fluorescence detection system.
  • Spectral resolution is achieved in the detection system by utilizing a dichroic mirror which serves to separate various wavelengths which are then separately detected by photomultiplier tubes.
  • the system additionally generates a reference beam which impinges on one of the scanning mirrors, the reflected scanning reference beam is directed through a grating and having an alternating sequence of transparent and opaque regions.
  • the transmitted beam is detected and utilized to generate a clock signal representative of the position of the scanning reference beam.
  • the clock signal is used to control analog-to-digital circuits in the fluorescence detection system. In this way, the sampling of the outputs of the photomultiplier tubes generates data representative of linear scans of the sample, despite the use of a scanning mirror that scans in a non-linear, sinusoidal fashion.
  • a scanning optical detection system provides for optical and mechanical positioning, alignment and examining of a sample.
  • a source of excitation radiation such as a laser, supplies excitation radiation to an optical detection platform either directly, or in the preferred embodiment, through a mechanically decoupled system such as an optical path, using optics or mirrors, or most preferably through an optical fiber.
  • the optical detection platform receives the excitation radiation, imparts a direction to the radiation, preferably through a x-y scanning system, examines the excitation radiation in the region to be examined, and detects emitted radiation from the object.
  • the detector preferably includes a filter adapted to substantially reject, preferably greater than a factor of 10 7 , excitation radiation.
  • the field of view of the detector preferably a photomultiplier tube, is of a restricted size, preferably restricted through an aperture disposed at the inlet to the photomultiplier tube.
  • the system comprises a confocal microscope system in which the excitation radiation illuminates one microlocation in an array of microlocations, but not other microlocations or intervening or interstitial areas at the same time.
  • the excitation radiation significantly illuminates a subset of the area comprising a microlocation.
  • the detector aperture is preferably sized to be of substantially the same lateral examining area as is the excitation radiation at the examined microlocation.
  • Improved methods of scanning utilizing a confocal optical detection system generally comprise the steps of, first, providing focused excitation energy to a region to be examined, that region comprising less than all of the region to be serially examined, second, focusing a detector on the region to be examined, the diameter of the detector aperture at the object to be examined being substantially the same as or smaller than the diameter of the object being examined, but the same as or greater than the diameter of the excitation radiation at the object to be examined, whereby the region of the object to be examined is illuminated and the detector is focused on the illuminated portion.
  • a dual detector system is utilized.
  • a first detector is operatively positioned to receive radiation reflected from the object, preferably including the microlocations, the first detector being coupled to a position detection system for determining the position of the microlocations.
  • a second detector is operatively positioned to receive the radiation emitted from the microlocations, such as fluorescent or chemiluminescent radiation.
  • Such a dual detector system is advantageously used in the methods for aligning the object including the microlocations to be examined with the optical detection platform.
  • An alignment system is provided for aligning the optical detection platform and the object to be examined.
  • an excitation radiation detector is used in combination with the scanning system and focusing optics.
  • the excitation radiation is scanned over the object to be examined, preferably through operation of the x-y positioning system, and the excitation radiation reflected from the object to be examined is made incident on the excitation detector.
  • the output of the excitation reflectance detector after association with the spatial coordinates available from the scanning system, can be used to extract optical information about the microlocations. If operated in a raster scanning mode, a two-dimensional image can be extracted from a single high sensitivity detector.
  • microlocation pattern is known, image processing techniques can be used to precisely determine the coordinates of the microlocations to the accuracy of the scanning system, which can be much more precise than the initial positioning of the microlocation bearing device in relation to the optical detection system. Once the position of the microlocations is known, the examining and detection of a specific microlocation may then be performed.
  • a laser power monitor is utilized. Both short and long term fluctuations in the power level of the excitation source may be corrected. Long term changes in the power level may be compensated for by changing the sensitivity of the detector, and short term fluctuations may be compensated for by multiplication of a correction factor applied to the output of the detector.
  • FIGS. 1 shows the active, programmable matrix system in perspective view.
  • FIG. 2 shows a plan view of multiple microlocations on an object to be examined.
  • FIG. 3 shows a block diagram of the system.
  • FIG. 4 shows a cross-sectional view of the optical detection platform and associated structures.
  • FIG. 5 shows a perspective view of the optical detection platform and associated components.
  • FIG. 6A shows a plan view of an array of microlocators with overlaid scans.
  • FIG. 6B shows the output of the excitation detector when scanning along line 240 a in FIG. 6A.
  • FIGS. 1 illustrates a simplified version of the active programmable electronic matrix (APEX) hybridization system for use with this invention.
  • a substrate 10 supports a matrix or array of electronically addressable microlocations 12 .
  • Relatively larger microlocations 16 may optionally be disposed around the smaller microlocations 12 .
  • the microlocations generally comprise those physical regions on or near the surface of the substrate 10 where some action or reaction of interest occurs, e.g., hybridization, ligand-anti-ligand reaction, which is later to be optically, e.g., via fluorescent or chemiluminescence, detected.
  • the active, programmable, matrix system transports charged material 14 to any of the specific microlocations 12 , such as the microlocation 12 labeled “+” in FIG. 1.
  • a microlocation as it relates to the detection system and methods of the instant inventions is generally characterized as being a substantially two-dimensional region, the two dimensions being preferably substantially parallel to the surface of the substrate 10 , the lateral extent of the microlocation typically being greater than the diffraction limited size of excitation radiation for use in the detection system.
  • a microlocation has a lateral dimension which is substantially greater, e.g., 5 times greater, and more preferably, 10 times greater than the lateral dimension of a diffraction limited spot size for the excitation source at the microlocation.
  • the microlocations may be separated by intervening or interstitial areas in which no observable reaction is intended to occur. However, microlocations need not be separated, such as in the case of contiguous microlocations.
  • FIG. 2 shows a plan view of an array of microlocations 20 to be examined. As shown, a 5 ⁇ 5 array of microlocations 20 is provided. While this number and arrangement of microlocations 20 is shown for convenience in FIG. 2, the number and positional arrangement of microlocations 20 relative to each other is unlimited.
  • Leads 22 connect a microlocation 20 to a power supply. As shown in FIG. 2, multiple leads 22 may be connected to a given microlocation 20 , though a single lead 22 may also be used to connect to a single microlocation 20 .
  • Electrodes 24 are disposed adjacent the array of microlocations 20 and are connected via one or more leads 26 to a power supply. As shown, the system typically includes interstitial regions 28 between the microlocations 20 .
  • the interstitial regions 28 comprise that space between the various microlocations 20 which contain the diagnostic or information bearing portions of the system.
  • the interstitial regions 28 are formed of material having low or reduced emission at the wavelength which corresponds to the emission wavelength, or is within the range of detection of the emission detector.
  • the array of microlocations is formed in an area nominally 1 cm ⁇ 1 cm.
  • the 5 ⁇ 5 array of microlocations 20 are within a 2 mm ⁇ 2 mm region.
  • An individual microlocation 20 may be of various diameters and shapes, but is preferably less than 100 ⁇ in diameter with the preferred shape being round.
  • the excitation beam and microlocation are both round.
  • FIG. 3 shows a block diagram view of the optical components of the system in association with a perspective view of an object bearing multiple microlocations to be examined.
  • An illumination, excitation source 40 preferably a laser
  • the optical block 44 passes the radiation from the source 40 to the scanning system 46 , which directs the radiation via objective lens assembly 48 towards the object to be examined 50 , which includes microlocations 12 disposed on a substrate 10 .
  • Light reflected from the object 50 including the microlocations 12 retraces through the objective lens assembly 48 , the scanning system 46 and enters the optical block 44 , where upon the reflectance detector 52 generates a reflectance signal 54 which is provided to the data acquisition system 66 .
  • a power monitor 58 generates a monitoring signal 60 , which constitutes a signal indicative of the power of the source 40 .
  • the power monitoring signal 60 is provides to the data acquisition system 66 .
  • Radiation from the excitation source 40 incident upon a microlocation 12 via the optical block 44 and scanning system 46 may, given a detectable condition, generate a detectable signal, such as a fluorescent or chemiluminescent radiation. Such emitted radiation passes via the scanning system 46 to the optical block 44 and to the aperture and focus assembly 62 , and the emitted radiation detector 64 .
  • the detector 64 preferably communicates with the control system 56 .
  • the detector 64 is optionally coupled to a data acquisition unit 66 .
  • a display 68 may be utilized to provide the user with a visual display.
  • a support 70 serves to support the substrate 10 .
  • FIG. 4 shows a cross-sectional view of the optical detection platform and associated devices.
  • An excitation source 160 provides illumination for the system.
  • a single laser source is used. While the excitation wavelength depends on the fluorophore, chromophore or other material to be excited, the preferred wavelength is 594 nm.
  • the diameter of the beam when incident on the surface 164 of the object to be examined is smaller than a given microlocation 162 (not to scale in FIG. 4).
  • various standards are known to the art for such a determination, such as the relative intensity falling to e ⁇ 2 .
  • the diameter of the beam is preferably nominal at 50 microns with the microlocations being at 80 microns.
  • Mode structure in the laser is preferably reduced by using a single mode laser and/or a single mode optical fiber.
  • the light 166 is transferred from the excitation source 160 to an optional fiber coupling 168 when an optical fiber 170 is used to deliver light 166 to the optical detection platform 172 .
  • the light 166 may also be passed through a filter 174 disposed in the optical path to eliminate spurious radiation from entering the optical detection platform 172 at a wavelength range other than that desired for excitation at the microlocation 162 .
  • the light 166 may be delivered to the optical detection platform 172 by other modes, whether by direct input from the excitation source or through the use of intervening optical elements and/or mirrors.
  • the excitation source 160 is mechanically decoupled from the optical detection platform 172 . Such decoupling advantageously permits easier replacement of the excitation source 160 and provides for greater stability of the optical detection platform 172 .
  • the excitation radiation 180 a is supplied to the optical detection platform 172 .
  • the excitation radiation will be labeled 180 a, 180 b, etc. to refer to sequential portions of the optical path.
  • Excitation radiation 180 a is first optionally provided to a first beamsplitter 182 where a reflected fraction of the excitation radiation 180 b is made incident on a laser power monitor 184 .
  • a transmitted portion of excitation radiation 180 c is passed through the first beam splitter 182 and optionally transmitted through a second beam splitter 186 to provide transmitted excitation radiation 180 d.
  • a dichroic beam splitter 188 provides a reflected excitation radiation 180 e towards the scanning system 190 .
  • the dichroic beam splitter 188 is made substantially totally reflective at the excitation wavelength and transmissive at the emission wavelength from the fluorophore, chromophore, or other wavelength to be detected from the microlocation.
  • the scanning system 190 may be of any form of beam placement system consistent with the goals and objects of this invention.
  • a two-axis, servocontrolled moveable mirror 192 imparts motion to the excitation radiation 180 e which is incident upon mirror 192 .
  • Motors 194 in combination with alignment screws 196 actuate contacts 198 bearing upon plate 200 which in turn moves mirror 192 .
  • Motion of the mirror 192 permits the selective directing of excitation radiation 180 e into excitation radiation 180 f which will be directed to a given microlocation 162 .
  • the use of a single mirror 192 permits the manufacture of a relatively smaller optical detection platform 172 as compared to a multiple mirror system and eliminates spatial distortion imparted by one axis upon another. Where size constraints are imposed upon the optical detection platform 172 , the single mirror 192 is preferred.
  • An objective lens 202 is disposed between the scanning system 190 and the object to be examined 164 and receives radiation 180 f and directs the radiation 180 g towards the microlocation 162 .
  • the objective lens 202 may be of any type known to those skilled in the art consistent with the goals and objects of this invention.
  • the objective lens 202 is an infinity corrected microscope lens. That is a lens designed to focus a collimated beam to a point, and vice versa.
  • the objective lens may be a commercially available microscope lens, or alternatively, constructed from one or more discrete lenses, such as those sold by Melles Griot.
  • the lens may be optimized as a scan lens, that is, a lens which has a linear relationship between the angle of the beam input and the position of the spot output. A relatively longer focal length scan lens permits scanning of a relatively larger area.
  • the primary optical path of the returning fluorescence 204 will be described, again using the convention of labeling 204 a, 204 b, etc. to refer to sequential portions of the optical path.
  • the emitted radiation 204 a from the microlocation 162 passes back, preferably, reversing the optical path of excitation radiation 180 g, 180 f and 180 e.
  • the region to be examined may be examined by imaging, or monitoring the emission intensity or otherwise by monitoring any parameter indicative of the biological event to be assayed or detected.
  • the emitted radiation 204 c is incident upon the dichroic beam splitter 188 , and is preferably substantially completely transmitted as emitted radiation 204 d.
  • Emitted radiation 204 d passes to a detector 208 , optionally through a tube 206 .
  • Detector 208 is chosen based upon the type and wavelength of emitted radiation 204 from the microlocations 162 .
  • the detector 208 is preferably a photomultiplier tube, most preferably one responsive in the range of from substantially 488 nm to substantially 800 nm.
  • a high sensitivity, low noise photomultiplier tube is preferred.
  • the photomultiplier tube 208 is operated in a current output mode utilizing a transconductance amplifier.
  • the photomultiplier tube 208 may be operated in a photon counting mode, with an integrator.
  • the emitted radiation 204 d is incident upon a filter 210 which serves to reject radiation at wavelengths which are not substantially the wavelength of the emitted radiation 204 .
  • the filter 210 should reject the excitation radiation 180 , preferably at least by a factor of 10 7 and more preferably by a factor of 10 10 .
  • the filtered emitted radiation 204 e is directed towards the detector 208 .
  • a receiving lens 209 serves to focus the radiation 204 .
  • the receiving lens 209 images the illuminated spot on the object to be examined 164 onto the plane of the aperture 212 .
  • an aperture 212 receives the emitted radiation 204 e.
  • the aperture 212 is preferably a pinhole aperture having a size such that the detector 208 receives light substantially only from a region not larger than, and preferably smaller than, the diameter of the microlocation 162 .
  • the actual aperture size depends on the magnification of the image, which is equal to the ratio of the focal lengths of the receiving lens 209 and the objective lens 202 . By way of example, if the receiving lens 209 has a focal length 3 times longer than the objective lens 202 , then the microlocation 162 will be magnified 3 times at the aperture 212 .
  • the aperture 212 would require a diameter of 180 microns.
  • the apparent size of the aperture 212 may be changed by moving it along the path of the emitted radiation 204 .
  • the aperture 212 limits the emitted radiation 204 from the examined microlocation 162 .
  • the location where the focus occurs moves with respect to the microlocations on the chip.
  • the aperture 212 is in focus at the microlocations on the chip, a relatively sharp cut-off of light emitted from outside of the aperture occurs.
  • the cut-off boundary is relatively larger, similar to the effect of a larger aperture. In this case, the cut-off is relatively less sharp, dropping relatively slowly past the out of focus boundary. Further, the collection efficiency from within the aperture image area is lessened.
  • the emitted radiation 204 f passing from the aperture 212 is supplied to detector 208 .
  • a focus motor 214 moves the detector 208 and aperture 212 . Movement of the aperture 212 permits optimization of the focus on the microlocations on the chip. Such an adjustment permits variations of the z position of the microlocations to be compensated for, thereby permitting more flexibility in the z axis positioning.
  • An optional alignment screw 216 serves to align the detector 208 with the remainder of the optical detection platform 172 .
  • a base 218 is preferably employed to provide support to the various components of the optical detection platform. Light baffles or other environmental modifying barriers may be utilized as desired.
  • the laser power monitor 184 detects the excitation radiation 180 b.
  • the power monitor 184 provides a signal indicative of the power level of the excitation source 160 .
  • Both short and long term fluctuations in the power level of the excitation source 160 may be corrected as necessary for proper examining and quantitation.
  • long term changes in the power level of the excitation source 160 may be compensated for by changing the sensitivity of the detector 208 , such as through changing the sensitivity of a photomultiplier tube.
  • Short term fluctuations in the power level of the excitation source 160 may be compensated through multiplication of a correction factor applied to the output of the detector 208 .
  • Accurate measurement of the laser power requires attention to the polarization states. While a conventional optical fiber may be utilized with a non-polarized laser, the use of a polarized laser in combination with a polarization preserving optical fiber is preferable to avoid polarization induced errors in power determinations.
  • FIG. 5 shows a cross-sectional view of the relationship of the optical detection platform (shown as the base plate 218 from the underside) and the objective lens 202 in relationship to various support components and examining components.
  • the cartridge 220 or other support for the microlocation to be examined is disposed on a support 70 (see e.g., FIG. 3) and is adapted for positioning within the field of view of the objective lens 202 .
  • a system is provided for mechanically positioning the cartridge 220 relative to the optical detection platform 172 .
  • Such a mechanical positioning could include a system such as shown in FIG. 5.
  • the cartridge 220 is optionally formed with multiple location points, such as a circular detent or opening 222 and slot 224 .
  • the opening 222 and slot 224 are formed at least on the upper surface, though may be formed through the cartridge 220 as shown.
  • the base 218 preferably includes pointed pins 226 and at least one, and preferably two, flat pin or pins 228 .
  • the pointed pins 226 are sized to coact with the circular opening 222 and slot 224 such that the pointed section of the pointed pin 226 indexes the cartridge 220 relative to the circular opening and has latitude in the wide direction in slot 224 .
  • the cartridge 220 may optionally be moveable in the x or y direction, preferably the y direction, to be removed from the overall system.
  • a heater is utilized to maintain the cartridge 220 at the desired temperature.
  • a cartridge 220 is presented to the overall system, preferably moving in the y direction into general position relative to base 218 .
  • the cartridge 220 moves in the z direction, resulting in mechanical alignment of the cartridge 220 relative to the base 218 by action of the circular opening 222 , slot 224 and the upper surface of the cartridge 220 in coaction with the pins 228 .
  • Such a system provides mechanical registration between the cartridge 220 and optical detection platform 172 . While a relatively high degree of alignment may be achieved through such a mechanical system, the optical alignment methods described herein are advantageously utilized to provide yet a higher level of precision alignment between the optical detection platform 172 and the microlocations 220 .
  • the optical detection platform 172 and associated components form, in the preferred embodiment, a confocal microscope system having a restricted or narrow excitation source where the diameter of the excitation source is substantially the same size or less than the diameter of a microlocation 162 (FIG. 4) in the array to be examined.
  • the excitation radiation 180 g is preferably in focus in the z-dimension at the microlocation 162 to be examined.
  • the emitted radiation 204 f to be received by the detector 208 is also of restricted or narrow aperture.
  • the lateral diameter of the microlocation examined as emitted radiation 204 f by the detector 208 is of substantially the same diameter as the microlocation, or more preferably less than the diameter of the microlocation 162 , and most preferably substantially the same as or less than the diameter of the excitation radiation 180 g on the microlocation 162 to be examined.
  • the sensitivity may be optimized by controlling the energy density of the excitation radiation and the intrinsic optical sensitivity of the detector.
  • the optical detection platform 172 may advantageously be utilized to provide information regarding the position of the microlocations 162 , interstitial regions 220 and, generally, the placement and positioning of the object to be examined 164 .
  • the excitation radiation 180 a may be supplied via, among others, the scanning system 190 , to multiple points on the surface of the object to be examined 164 .
  • the excitation radiation 180 h comprises excitation radiation 180 which has been reflected from the object to be examined 164 and detected at the excitation detector 240 (FIG. 4).
  • the multiple points are detected by scanning the excitation radiation 180 over the surface of the object to be examined 164 .
  • the received information may be used to form an image of the object to be examined 164 .
  • the received information from the excitation detector 240 is used in conjunction with preentered information regarding the relative position of the microlocations 162 and interstitial regions 220 . Since the structure of the object to be examined 164 is known prior to the alignment step, the amount of information required regarding the position of the object to be examined is reduced, and the positional determination may be made more rapidly as compared to the situation where the structure of the object to be examined 164 is unknown.
  • position may refer to absolute or relative position, for example, the values of the stepper motors 194 corresponding to a given mirror position may be considered a position (since they indicate where the microlocation is for purposes of illumination and detection).
  • FIG. 7A shows an array of microlocations 162 and interstitial areas 220 .
  • the scan lines 240 are shown over a portion of FIG. 6A, so as not to obscure the entire figure. Preferably, the entire area in which the array may be located in scanned. However, a lesser region may be scanned consistent with the goals and objects of this invention.
  • the array of microlocations 162 is oriented such that the scan line 240 A would scan the array along the long dimension of the array.
  • FIG. 6B shows the output from the detector 240 along one scan line (see FIG. 4). The scan in FIG. 6B generally shows relative intensity across the scan 240 A in FIG. 6A.
  • the position of the matrix may be determined. While the location of the microlocations 162 may be determined by examining the output of the detector 240 alone, it is advantageously utilized in conjunction with information regarding the structure of the device, such as the size and relative positioning of the microlocations 162 .
  • the system of this invention utilizes the optical detection platform 172 to both detect the fluorescence 204 from the object under investigation, as well as to detect the excitation radiation 180 h which is used to provide positioning information regarding the microlocations 162 .
  • the scanned excitation radiation 180 h is detected by the excitation detector 240 , which is provided to the detection system, which preferably in combination with the information regarding the positioning of the microlocations relative to one another, serves to direct the scanning system 190 to directly provide the excitation radiation 180 g to the microlocation 162 .
  • the positions of the microlocations 162 may be determined to a degree of precision sufficient to perform the fluorescence detection step by substantially illuminating only a desired microlocation.
  • the signal-to-noise ratio is increased from 10 4 to times 10 5 through use of a confocal system, reducing the area of illumination down to the desired imaging location may result in a reduction of scattered light to 1% or less compared to flood illumination, and imaging of that location provides yet another similar decrease in detected scattered radiation, resulting in a reduction of detected radiation from approximately ⁇ fraction (1/5000) ⁇ to ⁇ fraction (1/70,000) ⁇ .
  • the overall system parameters include that the detector 208 should have a minimum detectable fluorophore density of 0.2 fluorophores per square micron, a minimum optical signal-to-noise ratio of 10:1 at 1 second at 0.2 fluorophores per square micron, a maximum excitation energy of 0.1 microwatt per square micron and a detection resolution of 16 bits ⁇ 2, that is, a maximum decimal integer of 65,536 (2 16 ) for 4 states (2 2 ) for a resolution with a precision of ⁇ 4 parts out of 65,636, or ⁇ 0.006% of full scale.

Abstract

An optical detection system is adapted particularly for detection of biological reactions, especially fluorescent or chemilluminescent reactions. An excitation source, preferably a laser, illuminates a portion of an object to be examined, the portion preferably comprising one microlocation out of an array of microlocations. An intervening optical detection platform serves to direct the excitation radiation, preferably through use of a scanning system, most preferably through use of an x-y mirror-based scanning system to the portion of the object to be illuminated. A detector, preferably a photomultiplier tube, receives the emitted radiation from the objects to be examined, the detector being characterized in that the diameter of the region examined by the detector is the same as or smaller than the diameter of the illuminated region, and comprises less than the entire surface of the object to be examined, and most preferably images a whole or a part of a single microlocation. Preferably, the excitation source is coupled to the optical detection platform via an optical fiber. In operation, a confocal microscopy system is formed in which the excitation radiation is substantially in focus at the surface of the object to be examined, the excitation radiation having a lateral extent less than the entire diameter of the object to be examined and the detection system having a lateral field of view of a diameter substantially the same as or less than the diameter of the excitation region. In one aspect of this invention, the optical detection platform includes an excitation detector which measures reflected excitation radiation from the object to be examined. This information is then compared to prestored information regarding the location of the microlocations and interstitial regions on the object to be examined, whereby alignment information is obtained. The excitation radiation may then be precisely directed to a given microlocation or portion thereof so as to perform the examining through the confocal system. Significant increases in signal-to-noise ratio are achieve.

Description

    RELATED APPLICATION INFORMATION
  • This application is a continuation-in-part application of application Ser. No. 08/534,454, filed Sep. 27, 1995, entitled “Apparatus and Methods for Active Programmable Matrix Devices”, which is a continuation-in-part of application Ser. No. 08/304,657, filed Sep. 9, 1994, entitled, as amended, “Molecular Biological Diagnostic Systems Including Electrodes”, now allowed, which is a continuation-in-part of application Ser. No. 08/271,882, filed Jul. 7, 1994, entitled, as amended, “Methods for Electronic Stringency Control for Molecular Biological Analysis and Diagnostics”, now allowed, which is a continuation-in-part of application Ser. No. 07/146,504, filed Nov. 1, 1993, entitled, as amended, “Active Programmable Electronic Devices for Molecular Biological Analysis and Diagnostics”, now issued as U.S. Pat. No. 5,605,662, all incorporated herein by reference as if fully set forth herein.[0001]
    • FIELD OF THE INVENTION
  • This invention relates to optical detection and examining systems, especially systems for examining fluorescent or chemilluminescent radiation. More particularly, the invention relates to optical systems for examining localized areas containing biological fluorescent materials, where those systems require relatively high sensitivity. [0002]
    • BACKGROUND OF THE INVENTION
  • Molecular biology comprises a wide variety of techniques for the analysis of nucleic acid and protein. Many of these techniques and procedures form the basis of clinical diagnostic assays and tests. These techniques include nucleic acid hybridization analysis, restriction enzyme analysis, genetic sequence analysis, and the separation and purification of nucleic acids and proteins (See, e.g., J. Sambrook, E. F. Fritsch, and T. Maniatis, [0003] Molecular Cloning: A Laboratory Manual, 2 Ed., Cold spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989).
  • Most of these techniques involve carrying out numerous operations (e.g., pipetting, centrifugations, electrophoresis) on a large number of samples. They are often complex and time consuming, and generally require a high degree of accuracy. Many a technique is limited in its application by a lack of sensitivity, specificity, or reproducibility. For example, these problems have limited many diagnostic applications of nucleic acid hybridization analysis. [0004]
  • The complete process for carrying out a DNA hybridization analysis for a genetic or infectious disease is very involved. Broadly speaking, the complete process may be divided into a number of steps and substeps, broadly including the steps of obtaining the sample, disrupting the cells within the sample, performing complexity reduction or amplification, performing some sort of assay or hybridization, followed by detection of the presence or absence of a desired event serving to generate a result. [0005]
  • New techniques are being developed for carrying out multiple sample nucleic acid hybridization analysis on micro-formatted multiplex or matrix devices (e.g., DNA chips) (see M. Barinaga, 253 Science, pp. 1489, 1991; W. Bains, 10 Bio/Technology, pp. 757-758, 1992). These methods usually attach specific DNA sequences to very small specific areas of a solid support, such as micro-wells of a DNA chip. These hybridization formats are micro-scale versions of the conventional “dot blot” and “sandwich” hybridization systems. [0006]
  • A variety of methods exist for detection and analysis of the hybridization events. Depending on the reporter group (fluorophore, enzyme, radioisotope, etc.) used to label the DNA probe, detection and analysis are carried out fluorometrically, colorimetrically, or by autoradiography. By observing and measuring emitted radiation, such as fluorescent radiation or particle emission, information may be obtained about the hybridization events. Even when detection methods have very high intrinsic sensitivity, detection of hybridization events is difficult because of the background presence of non-specifically bound materials and materials with inherent fluorescent characteristics. A number of other factors also reduce the sensitivity and selectivity of DNA hybridization assays. [0007]
  • In conventional fluorometric detection systems, an excitation energy of one wavelength is delivered to the region of interest and energy of a different wavelength is emitted and detected. Large scale systems, generally those having a region of interest of two millimeters or greater, have been manufactured in which the quality of the overall system is not inherently limited by the size requirements of the optical elements or the ability to place them in optical proximity to the region of interest. However, with small geometries, such as those below 2 millimeters, and especially those on the order of 500 microns or less in size of the region of interest, the conventional approaches to fluorometer design have proved inadequate. Generally, the excitation and emission optical elements must be placed close to the region of interest. Preferably, a focused spot size is relatively small, often requiring sophisticated optical designs. As the size of the feature to be observed decreases, the demands for high accuracy in mechanical alignment increase. Further, because it is usually desirable to maximize the detectable area, the size of the optical components required to achieve these goals in relation to their distance from the region of interest becomes important, and in many cases, compromises the performance obtained. [0008]
  • Various prior art attempts have been made to image multiple sites in immunoassay systems. In Leaback, U.S. Pat. No. 5,096,807, there is a disclosure of an imaging immunoassay detection apparatus system and method purported to be capable of detecting and quantifying multiple light-emitting reactions from small volume samples simultaneously. A plurality of individual chemical reactant samples are each capable of emitting photons when a reaction takes place. These samples are arranged in a spaced relationship with respect to each other, and a detection system is operatively positioned so as to simultaneously detect the presence and x-y location of each photon emitted from any reacting sample. One disclosed carrier is a microtiter plate with multiple samples, e.g., 96, arranged in rows and columns. Various imaging devices are disclosed, such as an imaging photon detector, microchannel plate intensifiers and charged coupled devices (CCDs). Preferably, the signals representing the discrete areas of reactions have the background noise signal subtracted from them. [0009]
  • Yet other systems for imaging multiple sites in immunoassay systems utilize sequential scanning techniques. Multiple-well screening fluorometer systems move multiple sites relative to a fluorometer. Certain versions of the systems utilize a motorized stage and others arrange the samples on a wheel, which sequentially rotate samples into position for observation by the fluorometer. With these techniques, the samples are presented to the detector in a serial manner. [0010]
  • Another multiple location immunoassay system is disclosed in Elings et al. U.S. Pat. No. 4,537,861 entitled “Apparatus and Method for Homogeneous Immunoassay”. A spatial pattern formed by a spatial array of separate regions of antiligand material are disposed on a surface. The presence or absence of a binding reaction between a ligand and the antiligand is then detected. A source of illumination is shined on the combined ligand-antiligand location, and the emitted radiation detected. The contribution to the imager due to free labeled molecules plus background contaminants are suppressed through use of a chopper system in positional correlation to the examined array which generates a reference signal. [0011]
  • Various microscope systems for the detection of fluorescence or chemiluminescence have been known to the art. For example, Dixon et al. U.S. Pat. No. 5,192,980 entitled “Apparatus and Method for Spatially- and Spectrally- Resolved Measurements” discloses a scanning optical microscope or mapping system for spectrally-resolved measurement of light reflected, emitted or scattered from a specimen. A confocal scanning laser microscope system is combined with a grating monochromator located in the detector arm of the system. A spectrally resolved image is generated for a given point of illumination. Spatial resolution is achieved by moving the sample on a movable stage. [0012]
  • Another scanning confocal microscope is disclosed in U.S. Pat. No. 5,296,703 entitled “Scanning Confocal Microscope Using Fluorescence Detection”. A scanning confocal microscope is provided for scanning a sample with an incident beam of radiation and detecting the resulting fluorescence radiation to provide data suitable for use in a raster scanned display of the fluorescence. First and second closely spaced scanning mirrors direct an incident beam to a sample and direct the fluorescent radiation towards a fluorescence detection system. Spectral resolution is achieved in the detection system by utilizing a dichroic mirror which serves to separate various wavelengths which are then separately detected by photomultiplier tubes. The system additionally generates a reference beam which impinges on one of the scanning mirrors, the reflected scanning reference beam is directed through a grating and having an alternating sequence of transparent and opaque regions. The transmitted beam is detected and utilized to generate a clock signal representative of the position of the scanning reference beam. The clock signal is used to control analog-to-digital circuits in the fluorescence detection system. In this way, the sampling of the outputs of the photomultiplier tubes generates data representative of linear scans of the sample, despite the use of a scanning mirror that scans in a non-linear, sinusoidal fashion. [0013]
  • Despite the desirability of having an improved examining system, and the need for higher sensitivity in such systems, the systems described previously have been less than optimal. It is the object of this invention to provide an improved examining and scanning system which remedies these deficiencies. [0014]
    • SUMMARY OF THE INVENTION
  • A scanning optical detection system provides for optical and mechanical positioning, alignment and examining of a sample. A source of excitation radiation, such as a laser, supplies excitation radiation to an optical detection platform either directly, or in the preferred embodiment, through a mechanically decoupled system such as an optical path, using optics or mirrors, or most preferably through an optical fiber. The optical detection platform receives the excitation radiation, imparts a direction to the radiation, preferably through a x-y scanning system, examines the excitation radiation in the region to be examined, and detects emitted radiation from the object. The detector preferably includes a filter adapted to substantially reject, preferably greater than a factor of 10[0015] 7, excitation radiation. The field of view of the detector, preferably a photomultiplier tube, is of a restricted size, preferably restricted through an aperture disposed at the inlet to the photomultiplier tube.
  • In the preferred embodiment, the system comprises a confocal microscope system in which the excitation radiation illuminates one microlocation in an array of microlocations, but not other microlocations or intervening or interstitial areas at the same time. In the preferred embodiment, the excitation radiation significantly illuminates a subset of the area comprising a microlocation. Similarly, the detector aperture is preferably sized to be of substantially the same lateral examining area as is the excitation radiation at the examined microlocation. By restricting the scope of the illumination to the area of a given microlocation, or a fraction thereof, coupled with restricting the field of view of the detector to the region of illumination , preferably through use of an aperture, significant improvements in signal-to-noise ratio may be achieved. [0016]
  • Improved methods of scanning utilizing a confocal optical detection system generally comprise the steps of, first, providing focused excitation energy to a region to be examined, that region comprising less than all of the region to be serially examined, second, focusing a detector on the region to be examined, the diameter of the detector aperture at the object to be examined being substantially the same as or smaller than the diameter of the object being examined, but the same as or greater than the diameter of the excitation radiation at the object to be examined, whereby the region of the object to be examined is illuminated and the detector is focused on the illuminated portion. [0017]
  • In one aspect of this invention, a dual detector system is utilized. A first detector is operatively positioned to receive radiation reflected from the object, preferably including the microlocations, the first detector being coupled to a position detection system for determining the position of the microlocations. A second detector is operatively positioned to receive the radiation emitted from the microlocations, such as fluorescent or chemiluminescent radiation. Such a dual detector system is advantageously used in the methods for aligning the object including the microlocations to be examined with the optical detection platform. [0018]
  • An alignment system is provided for aligning the optical detection platform and the object to be examined. In the preferred embodiment, an excitation radiation detector is used in combination with the scanning system and focusing optics. In the preferred method of alignment, the excitation radiation is scanned over the object to be examined, preferably through operation of the x-y positioning system, and the excitation radiation reflected from the object to be examined is made incident on the excitation detector. The output of the excitation reflectance detector, after association with the spatial coordinates available from the scanning system, can be used to extract optical information about the microlocations. If operated in a raster scanning mode, a two-dimensional image can be extracted from a single high sensitivity detector. If the microlocation pattern is known, image processing techniques can be used to precisely determine the coordinates of the microlocations to the accuracy of the scanning system, which can be much more precise than the initial positioning of the microlocation bearing device in relation to the optical detection system. Once the position of the microlocations is known, the examining and detection of a specific microlocation may then be performed. [0019]
  • In yet another aspect of this invention, a laser power monitor is utilized. Both short and long term fluctuations in the power level of the excitation source may be corrected. Long term changes in the power level may be compensated for by changing the sensitivity of the detector, and short term fluctuations may be compensated for by multiplication of a correction factor applied to the output of the detector. [0020]
  • Accordingly, it is an object of this invention to provide an optical detection system having an improved signal-to-noise ratio. [0021]
  • It is yet another object of this invention to provide an examining system having high sensitivity and reliability. [0022]
  • It is yet another object of this invention to provide a sensitive diagnostic system at a relatively low cost.[0023]
    • BRIEF DESCRIPTION OF THE DRAWINGS
  • FIGS. [0024] 1 shows the active, programmable matrix system in perspective view.
  • FIG. 2 shows a plan view of multiple microlocations on an object to be examined. [0025]
  • FIG. 3 shows a block diagram of the system. [0026]
  • FIG. 4 shows a cross-sectional view of the optical detection platform and associated structures. [0027]
  • FIG. 5 shows a perspective view of the optical detection platform and associated components. [0028]
  • FIG. 6A shows a plan view of an array of microlocators with overlaid scans. [0029]
  • FIG. 6B shows the output of the excitation detector when scanning along line [0030] 240 a in FIG. 6A.
    • DETAILED DESCRIPTION OF THE INVENTION
  • FIGS. [0031] 1 illustrates a simplified version of the active programmable electronic matrix (APEX) hybridization system for use with this invention. Generally, a substrate 10 supports a matrix or array of electronically addressable microlocations 12. Relatively larger microlocations 16 may optionally be disposed around the smaller microlocations 12. The microlocations generally comprise those physical regions on or near the surface of the substrate 10 where some action or reaction of interest occurs, e.g., hybridization, ligand-anti-ligand reaction, which is later to be optically, e.g., via fluorescent or chemiluminescence, detected. In one mode of use, the active, programmable, matrix system transports charged material 14 to any of the specific microlocations 12, such as the microlocation 12 labeled “+” in FIG. 1.
  • A microlocation as it relates to the detection system and methods of the instant inventions is generally characterized as being a substantially two-dimensional region, the two dimensions being preferably substantially parallel to the surface of the [0032] substrate 10, the lateral extent of the microlocation typically being greater than the diffraction limited size of excitation radiation for use in the detection system. In the preferred embodiment, a microlocation has a lateral dimension which is substantially greater, e.g., 5 times greater, and more preferably, 10 times greater than the lateral dimension of a diffraction limited spot size for the excitation source at the microlocation. The microlocations may be separated by intervening or interstitial areas in which no observable reaction is intended to occur. However, microlocations need not be separated, such as in the case of contiguous microlocations.
  • FIG. 2 shows a plan view of an array of [0033] microlocations 20 to be examined. As shown, a 5×5 array of microlocations 20 is provided. While this number and arrangement of microlocations 20 is shown for convenience in FIG. 2, the number and positional arrangement of microlocations 20 relative to each other is unlimited. Leads 22 connect a microlocation 20 to a power supply. As shown in FIG. 2, multiple leads 22 may be connected to a given microlocation 20, though a single lead 22 may also be used to connect to a single microlocation 20. Electrodes 24 are disposed adjacent the array of microlocations 20 and are connected via one or more leads 26 to a power supply. As shown, the system typically includes interstitial regions 28 between the microlocations 20. The interstitial regions 28 comprise that space between the various microlocations 20 which contain the diagnostic or information bearing portions of the system. Preferably, the interstitial regions 28 are formed of material having low or reduced emission at the wavelength which corresponds to the emission wavelength, or is within the range of detection of the emission detector.
  • In the preferred embodiment, the array of microlocations is formed in an area nominally 1 cm×1 cm. In the embodiment shown, the 5×5 array of [0034] microlocations 20 are within a 2 mm×2 mm region. An individual microlocation 20 may be of various diameters and shapes, but is preferably less than 100μ in diameter with the preferred shape being round. In the preferred embodiment, the excitation beam and microlocation are both round.
  • FIG. 3 shows a block diagram view of the optical components of the system in association with a perspective view of an object bearing multiple microlocations to be examined. An illumination, [0035] excitation source 40, preferably a laser, provides radiation via a coupler assembly 42 to an optical block 44. The optical block 44 passes the radiation from the source 40 to the scanning system 46, which directs the radiation via objective lens assembly 48 towards the object to be examined 50, which includes microlocations 12 disposed on a substrate 10. Light reflected from the object 50 including the microlocations 12 retraces through the objective lens assembly 48, the scanning system 46 and enters the optical block 44, where upon the reflectance detector 52 generates a reflectance signal 54 which is provided to the data acquisition system 66. Optionally, a power monitor 58 generates a monitoring signal 60, which constitutes a signal indicative of the power of the source 40. The power monitoring signal 60 is provides to the data acquisition system 66.
  • Radiation from the [0036] excitation source 40 incident upon a microlocation 12 via the optical block 44 and scanning system 46 may, given a detectable condition, generate a detectable signal, such as a fluorescent or chemiluminescent radiation. Such emitted radiation passes via the scanning system 46 to the optical block 44 and to the aperture and focus assembly 62, and the emitted radiation detector 64. The detector 64 preferably communicates with the control system 56. The detector 64 is optionally coupled to a data acquisition unit 66. Further, a display 68 may be utilized to provide the user with a visual display. A support 70 serves to support the substrate 10.
  • FIG. 4 shows a cross-sectional view of the optical detection platform and associated devices. An [0037] excitation source 160 provides illumination for the system. In the preferred embodiment, a single laser source is used. While the excitation wavelength depends on the fluorophore, chromophore or other material to be excited, the preferred wavelength is 594 nm. Preferably, the diameter of the beam when incident on the surface 164 of the object to be examined is smaller than a given microlocation 162 (not to scale in FIG. 4). When the diameter of a beam is referred to, various standards are known to the art for such a determination, such as the relative intensity falling to e−2. For an APEX type device, the diameter of the beam is preferably nominal at 50 microns with the microlocations being at 80 microns. Mode structure in the laser is preferably reduced by using a single mode laser and/or a single mode optical fiber.
  • The light [0038] 166 is transferred from the excitation source 160 to an optional fiber coupling 168 when an optical fiber 170 is used to deliver light 166 to the optical detection platform 172. Optionally, the light 166 may also be passed through a filter 174 disposed in the optical path to eliminate spurious radiation from entering the optical detection platform 172 at a wavelength range other than that desired for excitation at the microlocation 162. Alternatively, the light 166 may be delivered to the optical detection platform 172 by other modes, whether by direct input from the excitation source or through the use of intervening optical elements and/or mirrors. Preferably, the excitation source 160 is mechanically decoupled from the optical detection platform 172. Such decoupling advantageously permits easier replacement of the excitation source 160 and provides for greater stability of the optical detection platform 172.
  • The excitation radiation [0039] 180 a is supplied to the optical detection platform 172. In FIG. 4, the excitation radiation will be labeled 180 a, 180 b, etc. to refer to sequential portions of the optical path. Excitation radiation 180 a is first optionally provided to a first beamsplitter 182 where a reflected fraction of the excitation radiation 180 b is made incident on a laser power monitor 184. A transmitted portion of excitation radiation 180 c is passed through the first beam splitter 182 and optionally transmitted through a second beam splitter 186 to provide transmitted excitation radiation 180 d. A dichroic beam splitter 188 provides a reflected excitation radiation 180 e towards the scanning system 190. Preferably, the dichroic beam splitter 188 is made substantially totally reflective at the excitation wavelength and transmissive at the emission wavelength from the fluorophore, chromophore, or other wavelength to be detected from the microlocation.
  • The [0040] scanning system 190 may be of any form of beam placement system consistent with the goals and objects of this invention. In the preferred embodiment, a two-axis, servocontrolled moveable mirror 192 imparts motion to the excitation radiation 180 e which is incident upon mirror 192. Motors 194 in combination with alignment screws 196 actuate contacts 198 bearing upon plate 200 which in turn moves mirror 192. Motion of the mirror 192 permits the selective directing of excitation radiation 180 e into excitation radiation 180 f which will be directed to a given microlocation 162. The use of a single mirror 192 permits the manufacture of a relatively smaller optical detection platform 172 as compared to a multiple mirror system and eliminates spatial distortion imparted by one axis upon another. Where size constraints are imposed upon the optical detection platform 172, the single mirror 192 is preferred.
  • An [0041] objective lens 202 is disposed between the scanning system 190 and the object to be examined 164 and receives radiation 180 f and directs the radiation 180 g towards the microlocation 162. The objective lens 202 may be of any type known to those skilled in the art consistent with the goals and objects of this invention. In the preferred embodiment, the objective lens 202 is an infinity corrected microscope lens. That is a lens designed to focus a collimated beam to a point, and vice versa. The objective lens may be a commercially available microscope lens, or alternatively, constructed from one or more discrete lenses, such as those sold by Melles Griot. Optionally, the lens may be optimized as a scan lens, that is, a lens which has a linear relationship between the angle of the beam input and the position of the spot output. A relatively longer focal length scan lens permits scanning of a relatively larger area.
  • The primary optical path of the returning fluorescence [0042] 204 will be described, again using the convention of labeling 204 a, 204 b, etc. to refer to sequential portions of the optical path. The emitted radiation 204 a from the microlocation 162 passes back, preferably, reversing the optical path of excitation radiation 180 g, 180 f and 180 e. As used herein, the region to be examined may be examined by imaging, or monitoring the emission intensity or otherwise by monitoring any parameter indicative of the biological event to be assayed or detected. The emitted radiation 204 c is incident upon the dichroic beam splitter 188, and is preferably substantially completely transmitted as emitted radiation 204 d. Emitted radiation 204 d passes to a detector 208, optionally through a tube 206. Detector 208 is chosen based upon the type and wavelength of emitted radiation 204 from the microlocations 162. In the case of an APEX device, where typically fluorescence is to be measured, the detector 208 is preferably a photomultiplier tube, most preferably one responsive in the range of from substantially 488 nm to substantially 800 nm. A high sensitivity, low noise photomultiplier tube is preferred. Preferably the photomultiplier tube 208 is operated in a current output mode utilizing a transconductance amplifier. Optionally, the photomultiplier tube 208 may be operated in a photon counting mode, with an integrator.
  • Optionally, the emitted radiation [0043] 204 d is incident upon a filter 210 which serves to reject radiation at wavelengths which are not substantially the wavelength of the emitted radiation 204. Most particularly, the filter 210 should reject the excitation radiation 180, preferably at least by a factor of 107 and more preferably by a factor of 1010. The filtered emitted radiation 204 e is directed towards the detector 208. A receiving lens 209 serves to focus the radiation 204. Preferably, the receiving lens 209 images the illuminated spot on the object to be examined 164 onto the plane of the aperture 212.
  • In the preferred embodiment, an [0044] aperture 212 receives the emitted radiation 204 e. The aperture 212 is preferably a pinhole aperture having a size such that the detector 208 receives light substantially only from a region not larger than, and preferably smaller than, the diameter of the microlocation 162. The actual aperture size depends on the magnification of the image, which is equal to the ratio of the focal lengths of the receiving lens 209 and the objective lens 202. By way of example, if the receiving lens 209 has a focal length 3 times longer than the objective lens 202, then the microlocation 162 will be magnified 3 times at the aperture 212. If the microlocation 162 is, e.g., 80 microns, to create a 60 micron diameter field of view for the detector 208, the aperture 212 would require a diameter of 180 microns.. The apparent size of the aperture 212 may be changed by moving it along the path of the emitted radiation 204. When the aperture 212 is at the focal point of the object lens 202, the aperture limits the emitted radiation 204 from the examined microlocation 162. As the aperture 212 is moved along the optic axis, the location where the focus occurs moves with respect to the microlocations on the chip. When the aperture 212 is in focus at the microlocations on the chip, a relatively sharp cut-off of light emitted from outside of the aperture occurs. If the system is not in focus, the cut-off boundary is relatively larger, similar to the effect of a larger aperture. In this case, the cut-off is relatively less sharp, dropping relatively slowly past the out of focus boundary. Further, the collection efficiency from within the aperture image area is lessened. The emitted radiation 204 f passing from the aperture 212 is supplied to detector 208.
  • Optionally, a [0045] focus motor 214 moves the detector 208 and aperture 212. Movement of the aperture 212 permits optimization of the focus on the microlocations on the chip. Such an adjustment permits variations of the z position of the microlocations to be compensated for, thereby permitting more flexibility in the z axis positioning. An optional alignment screw 216 serves to align the detector 208 with the remainder of the optical detection platform 172. A base 218 is preferably employed to provide support to the various components of the optical detection platform. Light baffles or other environmental modifying barriers may be utilized as desired.
  • The [0046] laser power monitor 184 detects the excitation radiation 180 b. The power monitor 184 provides a signal indicative of the power level of the excitation source 160. Both short and long term fluctuations in the power level of the excitation source 160 may be corrected as necessary for proper examining and quantitation. For example, long term changes in the power level of the excitation source 160 may be compensated for by changing the sensitivity of the detector 208, such as through changing the sensitivity of a photomultiplier tube. Short term fluctuations in the power level of the excitation source 160 may be compensated through multiplication of a correction factor applied to the output of the detector 208. Accurate measurement of the laser power requires attention to the polarization states. While a conventional optical fiber may be utilized with a non-polarized laser, the use of a polarized laser in combination with a polarization preserving optical fiber is preferable to avoid polarization induced errors in power determinations.
  • FIG. 5 shows a cross-sectional view of the relationship of the optical detection platform (shown as the [0047] base plate 218 from the underside) and the objective lens 202 in relationship to various support components and examining components. The cartridge 220 or other support for the microlocation to be examined is disposed on a support 70 (see e.g., FIG. 3) and is adapted for positioning within the field of view of the objective lens 202. In the preferred embodiment, a system is provided for mechanically positioning the cartridge 220 relative to the optical detection platform 172. Such a mechanical positioning could include a system such as shown in FIG. 5. The cartridge 220 is optionally formed with multiple location points, such as a circular detent or opening 222 and slot 224. The opening 222 and slot 224 are formed at least on the upper surface, though may be formed through the cartridge 220 as shown. One or more planar regions exist on the top of the cartridge 220. The base 218 preferably includes pointed pins 226 and at least one, and preferably two, flat pin or pins 228. The pointed pins 226 are sized to coact with the circular opening 222 and slot 224 such that the pointed section of the pointed pin 226 indexes the cartridge 220 relative to the circular opening and has latitude in the wide direction in slot 224. Additionally, the cartridge 220 may optionally be moveable in the x or y direction, preferably the y direction, to be removed from the overall system. In the preferred embodiment, a heater is utilized to maintain the cartridge 220 at the desired temperature.
  • In operation, a [0048] cartridge 220 is presented to the overall system, preferably moving in the y direction into general position relative to base 218. The cartridge 220 moves in the z direction, resulting in mechanical alignment of the cartridge 220 relative to the base 218 by action of the circular opening 222, slot 224 and the upper surface of the cartridge 220 in coaction with the pins 228. Such a system provides mechanical registration between the cartridge 220 and optical detection platform 172. While a relatively high degree of alignment may be achieved through such a mechanical system, the optical alignment methods described herein are advantageously utilized to provide yet a higher level of precision alignment between the optical detection platform 172 and the microlocations 220.
  • In operation, the [0049] optical detection platform 172 and associated components form, in the preferred embodiment, a confocal microscope system having a restricted or narrow excitation source where the diameter of the excitation source is substantially the same size or less than the diameter of a microlocation 162 (FIG. 4) in the array to be examined. The excitation radiation 180 g is preferably in focus in the z-dimension at the microlocation 162 to be examined. The emitted radiation 204 f to be received by the detector 208 is also of restricted or narrow aperture. Preferably the lateral diameter of the microlocation examined as emitted radiation 204 f by the detector 208 is of substantially the same diameter as the microlocation, or more preferably less than the diameter of the microlocation 162, and most preferably substantially the same as or less than the diameter of the excitation radiation 180 g on the microlocation 162 to be examined.
  • In the preferred embodiment, the combination of examining a microlocation through selective illumination by excitation radiation [0050] 180 g to a microlocation 162, but substantially not to interstitial regions 220 (see also interstitial regions 38 in FIG. 2) and by restricting the detection of the emitted radiation 204 f to the diameter of the microlocation, or more preferably to a diameter the same as or less than the diameter of the excitation radiation 180 g on the microlocation 162, the signal-to-noise ratio may be increased. The sensitivity may be optimized by controlling the energy density of the excitation radiation and the intrinsic optical sensitivity of the detector.
  • The [0051] optical detection platform 172 may advantageously be utilized to provide information regarding the position of the microlocations 162, interstitial regions 220 and, generally, the placement and positioning of the object to be examined 164. The excitation radiation 180 a may be supplied via, among others, the scanning system 190, to multiple points on the surface of the object to be examined 164. The excitation radiation 180 h comprises excitation radiation 180 which has been reflected from the object to be examined 164 and detected at the excitation detector 240 (FIG. 4). Preferably, the multiple points are detected by scanning the excitation radiation 180 over the surface of the object to be examined 164. By receiving, storing and comparing the excitation radiation 180 h as determined by the excitation detector 240, the received information may be used to form an image of the object to be examined 164. In the preferred embodiment, the received information from the excitation detector 240 is used in conjunction with preentered information regarding the relative position of the microlocations 162 and interstitial regions 220. Since the structure of the object to be examined 164 is known prior to the alignment step, the amount of information required regarding the position of the object to be examined is reduced, and the positional determination may be made more rapidly as compared to the situation where the structure of the object to be examined 164 is unknown. Once the position of the microlocations 162 relative to the optical detection platform 172 is known, the examining of a given microlocation 162 may be performed as described previously in connection with FIGS. 4 and 5. While the microlocations 162 are the preferred object to be imaged by the excitation radiation 180 h, other markers, alignment marks or fiducals may be utilized, alone or in combination, to form the imaging. When used herein, position may refer to absolute or relative position, for example, the values of the stepper motors 194 corresponding to a given mirror position may be considered a position (since they indicate where the microlocation is for purposes of illumination and detection).
  • FIGS. 6A and 6B show possible modes of scanning in the preferred method of alignment. FIG. 7A shows an array of [0052] microlocations 162 and interstitial areas 220. The scan lines 240 are shown over a portion of FIG. 6A, so as not to obscure the entire figure. Preferably, the entire area in which the array may be located in scanned. However, a lesser region may be scanned consistent with the goals and objects of this invention. In the preferred embodiment, the array of microlocations 162 is oriented such that the scan line 240A would scan the array along the long dimension of the array. FIG. 6B shows the output from the detector 240 along one scan line (see FIG. 4). The scan in FIG. 6B generally shows relative intensity across the scan 240A in FIG. 6A. By determining the periodicity, the position of the matrix may be determined. While the location of the microlocations 162 may be determined by examining the output of the detector 240 alone, it is advantageously utilized in conjunction with information regarding the structure of the device, such as the size and relative positioning of the microlocations 162.
  • In the preferred embodiment, the system of this invention utilizes the [0053] optical detection platform 172 to both detect the fluorescence 204 from the object under investigation, as well as to detect the excitation radiation 180 h which is used to provide positioning information regarding the microlocations 162. In this way, flexibility is provided regarding the mechanical positioning of the microlocations 162 relative to the remainder of the system. In the preferred mode, the scanned excitation radiation 180 h is detected by the excitation detector 240, which is provided to the detection system, which preferably in combination with the information regarding the positioning of the microlocations relative to one another, serves to direct the scanning system 190 to directly provide the excitation radiation 180 g to the microlocation 162. Through the use of the initial imaging step, the positions of the microlocations 162 may be determined to a degree of precision sufficient to perform the fluorescence detection step by substantially illuminating only a desired microlocation.
  • When utilized with an APEX system, the signal-to-noise ratio is increased from 10[0054] 4 to times 105 through use of a confocal system, reducing the area of illumination down to the desired imaging location may result in a reduction of scattered light to 1% or less compared to flood illumination, and imaging of that location provides yet another similar decrease in detected scattered radiation, resulting in a reduction of detected radiation from approximately {fraction (1/5000)} to {fraction (1/70,000)}. For use with the APEX device, the overall system parameters include that the detector 208 should have a minimum detectable fluorophore density of 0.2 fluorophores per square micron, a minimum optical signal-to-noise ratio of 10:1 at 1 second at 0.2 fluorophores per square micron, a maximum excitation energy of 0.1 microwatt per square micron and a detection resolution of 16 bits ±2, that is, a maximum decimal integer of 65,536 (216) for 4 states (22) for a resolution with a precision of ±4 parts out of 65,636, or ±0.006% of full scale.
  • The use of the dual detector system wherein the optical system is utilized to determine the positions of the microlocation and then, based upon that positional information, is utilized to provide excitation radiation to a given microlocation, provides significantly increased alignment characteristics relative to pure mechanical systems. Whereas the mechanical positioning in the combined system provides for a positioning accuracy of ±500 microns in the x and y directions, utilizing the optical position detection system of this invention permits an alignment accuracy of approximately 1 micron. [0055]
  • Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity and understanding, it will be readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims. [0056]

Claims (33)

We claim:
1. A system for optical detection of emitted radiation from microlocations on an object to be examined, comprising:
a light source providing excitation radiation characterized in that the diameter of the excitation radiation when illuminating the microlocation is of the same diameter or less than the diameter of the microlocation to be examined,
a confocal scanning system adapted to receive the excitation radiation and direct it to the microlocation and to provide reflected radiation and emitted radiation from the microlocation,
a first detector adapted to receive the reflected radiation and a position detection system, the first detector having an output connected to a position detection system,
a second detector operatively positioned to receive the emitted radiation from the microlocation, the detector characterized in that the diameter examined by the detector is less than or equal to the diameter of the excitation radiation, and
a control system coupled to the position detection system which causes the confocal scanning system to direct excitation radiation to a specific microlocation.
2. The system of
claim 1
 wherein the excitation source is a laser.
3. The system of
claim 2
 wherein the laser is a single laser source.
4. The system of
claim 1
 wherein the scanning system is an x-y scanning system.
5. The system of
claim 4
 wherein the x-y scanning system includes a mirror adapted to reflect the excitation radiation, the reflected radiation and the emitted radiation.
6. The system of
claim 1
 wherein the second detector further includes an aperture.
7. The system of
claim 6
 wherein the aperture comprises a pinhole aperture.
8. The system of
claim 7
 wherein the pinhole aperture corresponds to a microlocation with a diameter in the range from substantially 20 microns to 80 microns.
9. The system of
claim 8
 wherein the pinhole aperture corresponds to a microlocation with a diameter of substantially 50 microns.
10. The system of
claim 1
 wherein the second detector includes a photomultiplier tube.
11. The system of
claim 1
 further including a focusing motor.
12. The system of
claim 1
 further including a rejection filter disposed between the confocal scanning system and the second detector.
13. The system of
claim 12
 wherein the rejection filter rejects excitation radiation to a factor of 107.
14. The system of
claim 12
 wherein the rejection filter rejects excitation radiation to a factor of 1010.
15. The system of
claim 1
 further including a data acquisition system.
16. The system of
claim 1
 further including a display.
17. The system of
claim 1
 further including a laser power monitor positioned to receive excitation radiation and output an indication of laser power.
18. The system of
claim 17
 wherein the output of the laser power monitor is connected to the control system.
19. A confocal microscopy system including an optical detection platform comprising:
a restricted excitation source characterized in that the diameter of the excitation radiation incident upon the object to be examined is less than the lateral dimension of the object to be examined and at least substantially five times greater than the diffraction limited spot size of the excitation source, and
a detector characterized in that it has a restricted field of view in diameter which is no larger than the diameter of the excitation radiation incident upon the object to be examined.
20. The confocal microscopy system of
claim 19
 wherein the excitation source is a laser.
21. The confocal microscopy system of
claim 20
 wherein the diameter of the laser at the surface of the object to be examined is less than substantially 80 microns.
22. The confocal microscopy system of
claim 20
 wherein the diameter of the laser at the surface of the object to be examined is greater than substantially ten times the diffraction limited spot size of the excitation source.
23. The confocal microscopy system of
claim 20
 wherein the diameter of the laser at the surface of the object to be examined is greater than substantially 20 microns.
24. The confocal microscopy system of
claim 20
 wherein the diameter of the laser at the surface of the object to be examined is greater than substantially 40 microns.
25. The confocal microscopy system of
claim 20
 wherein the diameter of the laser at the surface of the object to be examined is approximately 50 microns.
26. The confocal microscopy system of
claim 20
 wherein the diameter of the laser at the surface of the object to be examined is in the range from 100 microns to 50 microns.
27. A method for optically examining a microlocation on an object comprising the steps of:
illuminating at least a portion of the object by scanning light from a source through a scanning confocal microscope onto the object, and detecting light reflected from the object through the scanning confocal microscope,
determining the position of the microlocation by analyzing the detected reflected light,
illuminating a microlocation with light from the source through the scanning confocal microscope utilizing the determined position of the microlocation, and
detecting emitted radiation from the microlocation.
28. The method of
claim 27
 for optically examining a microlocation wherein the step of determining the position of the microlocation includes use of information regarding the microlocation patterns.
29. The method of
claim 27
 for optically examining a microlocation wherein the step of illuminating the microlocation illuminates no more than a single microlocation.
30. The method of
claim 27
 for optically examining a microlocation wherein the step of detecting emitted radiation is subject to a field of view restricted to a microlocation.
31. A method for examining an object having multiple microlocations separated by interstitial areas comprising the steps of:
illuminating multiple points on the object to be examined,
detecting reflected radiation from the object to be examined,
comparing the information constituting reflected radiation with information regarding the structure of the object to be examined, whereby the position of the object is determined, and
illuminating one microlocation through a confocal microscope based upon the position information.
32. The method of
claim 31
 for alignment of an object having multiple microlocations wherein the multiple points of detection are gathered by scanning the radiation over at least two microlocations and one interstitial area.
33. A method for determining fluorescence intensity from multiple microlocations disposed on the surface of a biological diagnostic system comprising the steps of:
scanning the surface of the diagnostic system which includes the microlocations with a laser source directed through a scanning confocal optical system,
detecting light reflected from the microlocations, determining the position of the microlocations by imaging the reflected light,
illuminating one microlocation through the confocal optical system based upon the determined position, wherein the illumination does not extend substantially beyond the microlocation, and
detecting from the microlocation via the confocal microscope, where the detector masks emissions from the object in regions other than the one micro location.
US09/927,820 1993-11-01 2001-08-09 Scanning optical detection system Abandoned US20010052976A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US09/927,820 US20010052976A1 (en) 1993-11-01 2001-08-09 Scanning optical detection system

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US08/146,504 US5605662A (en) 1993-11-01 1993-11-01 Active programmable electronic devices for molecular biological analysis and diagnostics
US08/271,882 US6017696A (en) 1993-11-01 1994-07-07 Methods for electronic stringency control for molecular biological analysis and diagnostics
US08/304,657 US5632957A (en) 1993-11-01 1994-09-09 Molecular biological diagnostic systems including electrodes
US08/534,454 US5849486A (en) 1993-11-01 1995-09-27 Methods for hybridization analysis utilizing electrically controlled hybridization
US08/846,876 US6309601B1 (en) 1993-11-01 1997-05-01 Scanning optical detection system
US09/927,820 US20010052976A1 (en) 1993-11-01 2001-08-09 Scanning optical detection system

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
US08/846,876 Continuation US6309601B1 (en) 1993-11-01 1997-05-01 Scanning optical detection system

Publications (1)

Publication Number Publication Date
US20010052976A1 true US20010052976A1 (en) 2001-12-20

Family

ID=25299200

Family Applications (2)

Application Number Title Priority Date Filing Date
US08/846,876 Expired - Fee Related US6309601B1 (en) 1993-11-01 1997-05-01 Scanning optical detection system
US09/927,820 Abandoned US20010052976A1 (en) 1993-11-01 2001-08-09 Scanning optical detection system

Family Applications Before (1)

Application Number Title Priority Date Filing Date
US08/846,876 Expired - Fee Related US6309601B1 (en) 1993-11-01 1997-05-01 Scanning optical detection system

Country Status (3)

Country Link
US (2) US6309601B1 (en)
AU (1) AU7258398A (en)
WO (1) WO1998049543A1 (en)

Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020071359A1 (en) * 2000-12-08 2002-06-13 Worthington Mark Oscar Methods for detecting analytes using optical discs and optical disc readers
US20020118355A1 (en) * 2000-11-08 2002-08-29 Worthington Mark Oscar Interactive system for analyzing biological samples and processing related information and the use thereof
US20020171838A1 (en) * 2001-05-16 2002-11-21 Pal Andrew Attila Variable sampling control for rendering pixelization of analysis results in a bio-disc assembly and apparatus relating thereto
US20030052280A1 (en) * 2001-07-23 2003-03-20 Foster Thomas H. Method for operating a laser scanning confocal microscope system and a system thereof
US20030133840A1 (en) * 2001-10-24 2003-07-17 Coombs James Howard Segmented area detector for biodrive and methods relating thereto
US20050018583A1 (en) * 2000-12-08 2005-01-27 Worthington Mark O. Multiple data layer optical discs for detecting analytes
US20050046848A1 (en) * 2003-08-26 2005-03-03 Blueshift Biotechnologies, Inc. Time dependent fluorescence measurements
US20050264805A1 (en) * 2004-02-09 2005-12-01 Blueshift Biotechnologies, Inc. Methods and apparatus for scanning small sample volumes
WO2005124320A1 (en) * 2004-06-15 2005-12-29 Olympus Corporation Reaction container, and reaction device and detection device using the reaction container
US20060063160A1 (en) * 2004-09-22 2006-03-23 West Jay A Microfluidic microarray systems and methods thereof
US7141378B2 (en) 2004-07-02 2006-11-28 Blueshift Biotechnologies, Inc. Exploring fluorophore microenvironments
US7592139B2 (en) 2004-09-24 2009-09-22 Sandia National Laboratories High temperature flow-through device for rapid solubilization and analysis
US8974651B2 (en) 2010-04-17 2015-03-10 C.C. Imex Illuminator for visualization of fluorophores
US9835587B2 (en) 2014-04-01 2017-12-05 C.C. Imex Electrophoresis running tank assembly

Families Citing this family (78)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6129828A (en) 1996-09-06 2000-10-10 Nanogen, Inc. Apparatus and methods for active biological sample preparation
US6331274B1 (en) 1993-11-01 2001-12-18 Nanogen, Inc. Advanced active circuits and devices for molecular biological analysis and diagnostics
US5631734A (en) 1994-02-10 1997-05-20 Affymetrix, Inc. Method and apparatus for detection of fluorescently labeled materials
US7857957B2 (en) 1994-07-07 2010-12-28 Gamida For Life B.V. Integrated portable biological detection system
US6071394A (en) * 1996-09-06 2000-06-06 Nanogen, Inc. Channel-less separation of bioparticles on a bioelectronic chip by dielectrophoresis
US6780591B2 (en) 1998-05-01 2004-08-24 Arizona Board Of Regents Method of determining the nucleotide sequence of oligonucleotides and DNA molecules
US7875440B2 (en) 1998-05-01 2011-01-25 Arizona Board Of Regents Method of determining the nucleotide sequence of oligonucleotides and DNA molecules
US6818437B1 (en) * 1998-05-16 2004-11-16 Applera Corporation Instrument for monitoring polymerase chain reaction of DNA
US7498164B2 (en) 1998-05-16 2009-03-03 Applied Biosystems, Llc Instrument for monitoring nucleic acid sequence amplification reaction
JP2003524754A (en) * 1998-05-16 2003-08-19 ピーイー コーポレイション （エヌワイ） Apparatus for monitoring the polymerase chain reaction of DNA
US6838680B2 (en) 1999-05-12 2005-01-04 Aclara Biosciences, Inc. Multiplexed fluorescent detection in microfluidic devices
US6399952B1 (en) 1999-05-12 2002-06-04 Aclara Biosciences, Inc. Multiplexed fluorescent detection in microfluidic devices
US7423750B2 (en) * 2001-11-29 2008-09-09 Applera Corporation Configurations, systems, and methods for optical scanning with at least one first relative angular motion and at least one second angular motion or at least one linear motion
US7387891B2 (en) * 1999-05-17 2008-06-17 Applera Corporation Optical instrument including excitation source
US20050279949A1 (en) * 1999-05-17 2005-12-22 Applera Corporation Temperature control for light-emitting diode stabilization
US7410793B2 (en) 1999-05-17 2008-08-12 Applera Corporation Optical instrument including excitation source
FR2794861A1 (en) * 1999-06-11 2000-12-15 Commissariat Energie Atomique Device for reading a biochip comprises a light source for illuminating the biochip in the form of a strip covering at least one line of discrete zones
FR2797054A1 (en) * 1999-07-27 2001-02-02 Commissariat Energie Atomique DEVICE FOR READING A BIOLOGICAL CHIP
US6586257B1 (en) * 1999-10-12 2003-07-01 Vertex Pharmaceuticals Incorporated Multiwell scanner and scanning method
US6814933B2 (en) * 2000-09-19 2004-11-09 Aurora Biosciences Corporation Multiwell scanner and scanning method
US6642046B1 (en) * 1999-12-09 2003-11-04 Motorola, Inc. Method and apparatus for performing biological reactions on a substrate surface
US6379884B2 (en) * 2000-01-06 2002-04-30 Caliper Technologies Corp. Methods and systems for monitoring intracellular binding reactions
US6824669B1 (en) 2000-02-17 2004-11-30 Motorola, Inc. Protein and peptide sensors using electrical detection methods
US6358387B1 (en) 2000-03-27 2002-03-19 Caliper Technologies Corporation Ultra high throughput microfluidic analytical systems and methods
WO2001077645A1 (en) * 2000-04-05 2001-10-18 Siemens Production And Logistics Systems Ag Illuminating and imaging device, especially for carrying out quantitative fluorescence immuno tests
US6602400B1 (en) 2000-06-15 2003-08-05 Motorola, Inc. Method for enhanced bio-conjugation events
US7062092B2 (en) 2000-08-22 2006-06-13 Affymetrix, Inc. System, method, and computer software product for gain adjustment in biological microarray scanner
GB0102357D0 (en) * 2001-01-30 2001-03-14 Randox Lab Ltd Imaging method
US6650411B2 (en) 2001-04-26 2003-11-18 Affymetrix, Inc. System, method, and product for pixel clocking in scanning of biological materials
US6643015B2 (en) 2001-04-26 2003-11-04 Affymetrix, Inc. System, method, and product for symmetrical filtering in scanning of biological materials
US6490533B2 (en) 2001-04-26 2002-12-03 Affymetrix, Inc. System, method, and product for dynamic noise reduction in scanning of biological materials
DE10125469B4 (en) * 2001-05-25 2008-01-10 Leica Microsystems Cms Gmbh Device for determining a light output, microscope and method for microscopy
US7635588B2 (en) * 2001-11-29 2009-12-22 Applied Biosystems, Llc Apparatus and method for differentiating multiple fluorescence signals by excitation wavelength
US6887362B2 (en) * 2002-02-06 2005-05-03 Nanogen, Inc. Dielectrophoretic separation and immunoassay methods on active electronic matrix devices
CA2422224A1 (en) 2002-03-15 2003-09-15 Affymetrix, Inc. System, method, and product for scanning of biological materials
ATE537438T1 (en) * 2002-05-17 2011-12-15 Life Technologies Corp DEVICE AND METHOD FOR DISTINGUISHING SEVERAL FLUORESCENCE SIGNALS BASED ON THEIR EXCITATION WAVELENGTH
US7164533B2 (en) 2003-01-22 2007-01-16 Cyvera Corporation Hybrid random bead/chip based microarray
US7900836B2 (en) 2002-08-20 2011-03-08 Illumina, Inc. Optical reader system for substrates having an optically readable code
US7901630B2 (en) 2002-08-20 2011-03-08 Illumina, Inc. Diffraction grating-based encoded microparticle assay stick
US7508608B2 (en) 2004-11-17 2009-03-24 Illumina, Inc. Lithographically fabricated holographic optical identification element
US7923260B2 (en) 2002-08-20 2011-04-12 Illumina, Inc. Method of reading encoded particles
US7872804B2 (en) 2002-08-20 2011-01-18 Illumina, Inc. Encoded particle having a grating with variations in the refractive index
US20100255603A9 (en) 2002-09-12 2010-10-07 Putnam Martin A Method and apparatus for aligning microbeads in order to interrogate the same
US7092160B2 (en) 2002-09-12 2006-08-15 Illumina, Inc. Method of manufacturing of diffraction grating-based optical identification element
EP2096341A1 (en) * 2002-12-04 2009-09-02 Spinx, Inc, Devices and methods for programmable microscale manipulation of fluids
US7265829B2 (en) * 2002-12-17 2007-09-04 Molecular Devices Corporation Reflective optic system for imaging microplate readers
US20040150217A1 (en) * 2003-01-23 2004-08-05 Heffelfinger David M. Identifying indicia and focusing target
US7317415B2 (en) 2003-08-08 2008-01-08 Affymetrix, Inc. System, method, and product for scanning of biological materials employing dual analog integrators
US7169560B2 (en) 2003-11-12 2007-01-30 Helicos Biosciences Corporation Short cycle methods for sequencing polynucleotides
TWI240988B (en) * 2004-01-07 2005-10-01 Powerchip Semiconductor Corp Method for fabricating a through hole on a semiconductor substrate
EP2248911A1 (en) 2004-02-19 2010-11-10 Helicos Biosciences Corporation Methods and kits for analyzing polynucleotide sequences
US7433123B2 (en) 2004-02-19 2008-10-07 Illumina, Inc. Optical identification element having non-waveguide photosensitive substrate with diffraction grating therein
WO2006020363A2 (en) 2004-07-21 2006-02-23 Illumina, Inc. Method and apparatus for drug product tracking using encoded optical identification elements
WO2006055736A1 (en) 2004-11-16 2006-05-26 Illumina, Inc. And methods and apparatus for reading coded microbeads
US7768638B2 (en) * 2005-03-18 2010-08-03 Illumina, Inc. Systems for and methods of facilitating focusing an optical scanner
US8351026B2 (en) 2005-04-22 2013-01-08 Affymetrix, Inc. Methods and devices for reading microarrays
US7666593B2 (en) 2005-08-26 2010-02-23 Helicos Biosciences Corporation Single molecule sequencing of captured nucleic acids
US7329860B2 (en) 2005-11-23 2008-02-12 Illumina, Inc. Confocal imaging methods and apparatus
US7830575B2 (en) 2006-04-10 2010-11-09 Illumina, Inc. Optical scanner with improved scan time
US8009889B2 (en) 2006-06-27 2011-08-30 Affymetrix, Inc. Feature intensity reconstruction of biological probe array
US7791013B2 (en) * 2006-11-21 2010-09-07 Illumina, Inc. Biological microarray line scanning method and system
US7813013B2 (en) * 2006-11-21 2010-10-12 Illumina, Inc. Hexagonal site line scanning method and system
DK2156370T3 (en) 2007-05-14 2012-01-23 Historx Inc Compartment separation by pixel characterization using image data clustering
EP2162728B1 (en) * 2007-06-15 2016-07-27 Novartis AG Microscope system and method for obtaining standardized sample data
EP2335221B8 (en) 2008-09-16 2016-05-25 Novartis AG Reproducible quantification of biomarker expression
JP5715068B2 (en) 2009-01-30 2015-05-07 マイクロニクス， インコーポレイテッド Portable high gain fluorescence detection system
US9767342B2 (en) 2009-05-22 2017-09-19 Affymetrix, Inc. Methods and devices for reading microarrays
WO2011094577A2 (en) 2010-01-29 2011-08-04 Micronics, Inc. Sample-to-answer microfluidic cartridge
JP5817378B2 (en) * 2011-01-28 2015-11-18 東レ株式会社 Microarray analysis method and reader
WO2014100732A1 (en) 2012-12-21 2014-06-26 Micronics, Inc. Fluidic circuits and related manufacturing methods
JP2016509206A (en) 2012-12-21 2016-03-24 マイクロニクス， インコーポレイテッド Portable fluorescence detection system and microassay cartridge
KR20150096788A (en) 2012-12-21 2015-08-25 마이크로닉스 인코포레이티드. Low elasticity films for microfluidic use
US10386377B2 (en) 2013-05-07 2019-08-20 Micronics, Inc. Microfluidic devices and methods for performing serum separation and blood cross-matching
WO2014182847A1 (en) 2013-05-07 2014-11-13 Micronics, Inc. Device for preparation and analysis of nucleic acids
US10190153B2 (en) 2013-05-07 2019-01-29 Micronics, Inc. Methods for preparation of nucleic acid-containing samples using clay minerals and alkaline solutions
US10571676B1 (en) 2018-08-09 2020-02-25 Viavi Solutions Inc. Determining an error in a moving distance of a microscope
US11231571B2 (en) * 2018-08-09 2022-01-25 Viavi Solutions Inc. Determining an erroneous movement of a microscope
WO2022056385A1 (en) 2020-09-14 2022-03-17 Singular Genomics Systems, Inc. Methods and systems for multidimensional imaging

Family Cites Families (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4572668A (en) 1982-08-26 1986-02-25 Midac Corporation Apparatus and method for photoluminescent analysis
US4555731A (en) 1984-04-30 1985-11-26 Polaroid Corporation Electronic imaging camera with microchannel plate
US5096807A (en) 1985-03-06 1992-03-17 Murex Corporation Imaging immunoassay detection system with background compensation and its use
US4707235A (en) 1985-10-16 1987-11-17 Pharmacia, Inc. Electrophoresis method and apparatus having continuous detection means
US4835704A (en) * 1986-12-29 1989-05-30 General Electric Company Adaptive lithography system to provide high density interconnect
JPH0799353B2 (en) 1987-03-31 1995-10-25 株式会社島津製作所 Nucleotide sequencer
US4827125A (en) * 1987-04-29 1989-05-02 The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Confocal scanning laser microscope having no moving parts
IL89874A0 (en) * 1989-04-06 1989-12-15 Nissim Nejat Danon Apparatus for computerized laser surgery
GB9014263D0 (en) 1990-06-27 1990-08-15 Dixon Arthur E Apparatus and method for spatially- and spectrally- resolvedmeasurements
US5296703A (en) 1992-04-01 1994-03-22 The Regents Of The University Of California Scanning confocal microscope using fluorescence detection
US5324401A (en) 1993-02-05 1994-06-28 Iowa State University Research Foundation, Inc. Multiplexed fluorescence detector system for capillary electrophoresis
US5381224A (en) 1993-08-30 1995-01-10 A. E. Dixon Scanning laser imaging system
US5578832A (en) 1994-09-02 1996-11-26 Affymetrix, Inc. Method and apparatus for imaging a sample on a device
US5631734A (en) 1994-02-10 1997-05-20 Affymetrix, Inc. Method and apparatus for detection of fluorescently labeled materials
US5545531A (en) * 1995-06-07 1996-08-13 Affymax Technologies N.V. Methods for making a device for concurrently processing multiple biological chip assays

Cited By (26)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6937323B2 (en) 2000-11-08 2005-08-30 Burstein Technologies, Inc. Interactive system for analyzing biological samples and processing related information and the use thereof
US20020118355A1 (en) * 2000-11-08 2002-08-29 Worthington Mark Oscar Interactive system for analyzing biological samples and processing related information and the use thereof
US7110345B2 (en) 2000-12-08 2006-09-19 Burstein Technologies, Inc. Multiple data layer optical discs for detecting analytes
US6995845B2 (en) 2000-12-08 2006-02-07 Burstein Technologies, Inc. Methods for detecting analytes using optical discs and optical disc readers
US20050018583A1 (en) * 2000-12-08 2005-01-27 Worthington Mark O. Multiple data layer optical discs for detecting analytes
US20020071359A1 (en) * 2000-12-08 2002-06-13 Worthington Mark Oscar Methods for detecting analytes using optical discs and optical disc readers
US20020171838A1 (en) * 2001-05-16 2002-11-21 Pal Andrew Attila Variable sampling control for rendering pixelization of analysis results in a bio-disc assembly and apparatus relating thereto
US20050110994A1 (en) * 2001-07-23 2005-05-26 Foster Thomas H. Method for operating a laser scanning confocal microscope system and a system thereof
US6956647B2 (en) 2001-07-23 2005-10-18 University Of Rochester Method for operating a laser scanning confocal microscope system and a system thereof
US6859273B2 (en) 2001-07-23 2005-02-22 University Of Rochester Method for operating a laser scanning confocal microscope system and a system thereof
US20030052280A1 (en) * 2001-07-23 2003-03-20 Foster Thomas H. Method for operating a laser scanning confocal microscope system and a system thereof
US20030133840A1 (en) * 2001-10-24 2003-07-17 Coombs James Howard Segmented area detector for biodrive and methods relating thereto
US20080174842A1 (en) * 2003-08-26 2008-07-24 Blueshift Biotechnologies, Inc. Time dependent fluorescence measurements
US20050046848A1 (en) * 2003-08-26 2005-03-03 Blueshift Biotechnologies, Inc. Time dependent fluorescence measurements
US7812956B2 (en) 2003-08-26 2010-10-12 Blueshift Biotechnologies, Inc. Time dependent fluorescence measurements
US7576862B2 (en) 2003-08-26 2009-08-18 Blueshift Biotechnologies, Inc. Measuring time dependent fluorescence
US20050264805A1 (en) * 2004-02-09 2005-12-01 Blueshift Biotechnologies, Inc. Methods and apparatus for scanning small sample volumes
WO2005124320A1 (en) * 2004-06-15 2005-12-29 Olympus Corporation Reaction container, and reaction device and detection device using the reaction container
US7141378B2 (en) 2004-07-02 2006-11-28 Blueshift Biotechnologies, Inc. Exploring fluorophore microenvironments
US7524672B2 (en) 2004-09-22 2009-04-28 Sandia Corporation Microfluidic microarray systems and methods thereof
US20060063160A1 (en) * 2004-09-22 2006-03-23 West Jay A Microfluidic microarray systems and methods thereof
US7592139B2 (en) 2004-09-24 2009-09-22 Sandia National Laboratories High temperature flow-through device for rapid solubilization and analysis
US8426135B1 (en) 2004-09-24 2013-04-23 Sandia National Laboratories High temperature flow-through device for rapid solubilization and analysis
US8974651B2 (en) 2010-04-17 2015-03-10 C.C. Imex Illuminator for visualization of fluorophores
US9835587B2 (en) 2014-04-01 2017-12-05 C.C. Imex Electrophoresis running tank assembly
US10641731B2 (en) 2014-04-01 2020-05-05 C.C. Imex Electrophoresis running tank assembly

Also Published As

Publication number Publication date
WO1998049543A1 (en) 1998-11-05
US6309601B1 (en) 2001-10-30
AU7258398A (en) 1998-11-24

Similar Documents

Publication Publication Date Title
US6309601B1 (en) Scanning optical detection system
US6704104B2 (en) Multi-wavelength array reader for biological assay
US20030112432A1 (en) Apparatus for reading signals generated from resonance light scattered particle labels
US7198939B2 (en) Apparatus for interrogating an addressable array
US6750457B2 (en) System for high throughput analysis
US7170597B1 (en) Microplate reader
EP1681556B1 (en) Imaging fluorescence signals using telecentricity
US7199377B2 (en) Optical analytic measurement device for fluorescence measurements in multisample carriers
US7551271B2 (en) Uncaging devices
US20040219536A1 (en) Chemical array reading
US20080068602A1 (en) Device for Reading Plates Bearing Biological Reaction Support Microdepositions
CN101868752A (en) Optical illumination apparatus for illuminating a sample with a line beam
US20020005493A1 (en) Optical components for microarray analysis
JP2002005834A (en) Distribution measuring apparatus for fluorescence labeled substance
US20080253409A1 (en) Multi-Channel Bio-Chip Scanner
EP2225548B1 (en) Detection system and method
WO2001057501A1 (en) Improved microarray reader
JP4967261B2 (en) Probe carrier
Visalli et al. Microarrays as a Tool for Gene Expression Profiling: Application in Ocular and kk

Legal Events

Date Code Title Description
STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION