WO1997039144A1 - Dispositif optique integre de detection moleculaire et procede correspondant - Google Patents

Dispositif optique integre de detection moleculaire et procede correspondant Download PDF

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Publication number
WO1997039144A1
WO1997039144A1 PCT/US1997/005534 US9705534W WO9739144A1 WO 1997039144 A1 WO1997039144 A1 WO 1997039144A1 US 9705534 W US9705534 W US 9705534W WO 9739144 A1 WO9739144 A1 WO 9739144A1
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WO
WIPO (PCT)
Prior art keywords
substrate
binding site
integrated
optical
molecular
Prior art date
Application number
PCT/US1997/005534
Other languages
English (en)
Inventor
Richard Ackley
Chan-Long Shieh
Original Assignee
Motorola 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
Application filed by Motorola Inc. filed Critical Motorola Inc.
Priority to AU24365/97A priority Critical patent/AU2436597A/en
Publication of WO1997039144A1 publication Critical patent/WO1997039144A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
    • 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
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator

Definitions

  • the present invention relates to optical molecular detection devices and methods of detection therefor.
  • a molecular detection chip includes a substrate on which an array of binding sites is arranged. Each binding site (or hybridization site) has a respective molecular receptor which binds or hybridizes with a molecule having a predetermined structure.
  • a sample solution is applied to the molecular detection chip, and molecules in the sample bind or hybridize at one or more of the binding sites. The particular binding sites at which hybridization occurs are detected, and one or more molecular structures within the sample are subsequently deduced.
  • DNA chips utilize an array of binding sites each having respective single-stranded DNA probes.
  • a sample of single- stranded DNA fragments referred to as target DNA, is applied to the DNA chip.
  • the DNA fragments attach to one or more of the DNA probes by a hybridization process. By detecting which DNA probes have a DNA fragment hybridized thereto, a sequence of nucleotide bases within the DNA fragment can be determined.
  • a local concentration of target DNA can be increased at predetermined sites using electric field enhancements.
  • each site has an electrode associated therewith for selectively generating an electric field thereby.
  • the electric field is generated by applying an electric potential between an electrode at the site and a counter electrode at a peripheral portion of the chip.
  • the polarity of the electric potential is selected to generate an electric field having a polarity opposite to the charge of the DNA fragments.
  • an electric field having the same polarity as the DNA fragments can be generated to repel the DNA fragments from the site.
  • a radioactive marker is attached to each of a plurality of molecules in the sample.
  • the binding of a molecule to a molecular receptor is then detectable by detecting the radioactive marker.
  • fluorescent labels such as fluorophores which selectively illuminate when hybridization occurs. These fluorophores are illuminated by a pump light source external to the substrate. An external charge-coupled device (CCD) camera is utilized to detect fluorescence from the illuminated fluorophores.
  • CCD charge-coupled device
  • An optical-detection-based DNA chip is disclosed in "Real-time detection of DNA hybridization and melting on oligonucleotide arrays by using optical wave guides", Proceedings of the National Academy of Sciences, Vol. 92, pp. 6379-6383. This DNA chip utilizes a glass substrate having a surface which defines the binding sites. The glass substrate has an end adjacent to the surface into which light is injected by an external light source. An external CCD camera is utilized to detect scatter light at the binding sites at which hybridization occurs.
  • FIG. 1 is a block diagram of an embodiment of an integrated molecular detection device in accordance with the present invention
  • FIG. 2 is a schematic cross-sectional view of an embodiment of an integrated molecular detection device in accordance with the present invention
  • FIG. 3 is a flow chart of an embodiment of a method of detecting hybridization of a molecule to a molecular receptor
  • FIG. 4 is a schematic perspective view of a preferred embodiment of an integrated molecular detection chip in accordance with the present invention.
  • FIG. 5 is a schematic cross-sectional view of the preferred embodiment of the integrated molecular detection chip. Detailed Description of a Preferred Embodiment
  • Embodiments of the present invention advantageously provide an integrated molecular detection device that incorporates binding sites, a light source, and optical detection electronics onto a single chip.
  • a preferred embodiment utilizes a micromachined glass substrate, a polymer light-emitting diode light source, and a thin-film- transistor-based optical imager to provide a fully integrated chip.
  • the device can perform field-enhanced hybridization and de-hybridization.
  • Embodiments of the present invention can be utilized for DNA sequencing and detection applications.
  • FIG. 1 is a block diagram of an embodiment of an integrated molecular detection device 10 in accordance with the present invention.
  • the integrated molecular detection device 10 includes a substrate 12 which defines a binding site 14 for receiving a molecular receptor 16.
  • the molecular receptor 16 is selected in dependence upon the type of molecule which is to be detected.
  • the molecular receptor 16 typically includes a biological or synthetic molecule that has a specific affinity to the molecule to be detected.
  • the molecular receptor 16 includes at least one DNA probe having a specific base pair sequence. It is noted that embodiments of the present invention are not limited to detecting hybridization of DNA molecules. For example, embodiments of the present invention can be utilized to detect antibody-antigen binding events.
  • a light source 18 is integrated in the substrate 12.
  • the light source 18 illuminates the binding site 14 through the substrate 12 to produce an optical indication of a molecule 19 hybridized or bound to the molecular receptor 16.
  • the optical indication can be provided, for example, by a fluorophore that selectively illuminates when hybridization occurs.
  • An optical detector 20 is also integrated in the substrate 12.
  • the optical detector 20 detects the optical indication through the substrate 12, and hence detects the hybridization or the binding of the molecule 19.
  • the substrate 12 is formed of glass so that the binding site 14 can be illuminated through the substrate 12, and so the optical indication can be detected through the substrate 12.
  • the substrate 12 can be formed of a substantially-transparent material to allow illumination and detection therethrough.
  • FIG. 2 is a schematic cross-sectional view of another embodiment of an integrated molecular detection device in accordance with the present invention.
  • the integrated molecular detection device includes a substrate 30 which can be formed of glass.
  • the substrate 30 defines a first side 32 (or a top side) and a second side 34 (or a reverse side) . As illustrated, the first side 32 opposes the second side 34 in this embodiment .
  • a binding site 36 for receiving a molecular receptor 37 is defined on the first side 32 of the substrate 30.
  • the binding site 36 can be formed by etching or micromachining a well into the substrate 30.
  • the molecular receptor 37 is deposited into the binding site 36 using a robotic dispensing technique, a self-assembly technique, or other techniques known in the art.
  • the molecular receptor 37 includes a chain of at least one nucleotide for detecting a molecule having a complementary chain of at least one nucleotide.
  • the molecular receptor 37 can include at least one DNA probe which selectively hybridizes with predetermined DNA molecules.
  • a light source 38 is incorporated on the second side 34 of the substrate 30.
  • the light source 38 includes a polymer light-emitting diode fabricated on the second side 34 of the substrate 30.
  • the binding site 36 is illuminated by the light source 38 through the substrate 30.
  • light generated at the second side 34 of the substrate 30 (by the light source 38) is transmitted through the substrate 30 to the binding site 36 at the first side 32.
  • Illuminating the binding site 36 acts to illuminate an optical indicator of a molecule which has hybridized to the molecular receptor 37.
  • Illuminating the optical indicator acts to produce an optical indication of the hybridization.
  • the optical indicator can be provided by a fluorophore that selectively activates when hybridization occurs.
  • the light source 38 preferably has an spectral output corresponding to absorption bands of the fluorophore.
  • the number of fluorescent molecules can be increased by attaching multiple fluorescent molecules to a bead or like member which is attached to the molecule. It is noted, however, that embodiments of the present invention are not limited to the use of fluorescent indicators, and that other optical indicators can be utilized for the same purpose.
  • An optical detector 40 is incorporated on the second side 34 of the substrate 30 to detect the optical indication.
  • the optical detector 40 can include a photodiode fabricated on the second side 34 of the substrate 30.
  • the optical detector 40 detects the optical indication, and hence detects the hybridization of the molecule, through the substrate 30.
  • the optical indication generated at the first side 32 of the substrate 30 is transmitted through the substrate 30 to the optical detector 40 at the second side 34.
  • the light source 38 and the optical detector 40 are located beside one another on the second side 34 of the substrate 30.
  • the integrated molecular detection device includes a filter 42 incorporated over the optical detector 40.
  • the filter 42 can include a color filter to screen out fluorescent light emanating from the light source 38.
  • the integrated molecular detection device includes a transparent electrode 44 incorporated in the substrate 30 at a location proximate to the binding site 36.
  • the transparent electrode 44 is utilized for field- assisted hybridization and de-hybridization of the molecule with the molecular receptor 37. Because of its transparency, the transparent electrode 44 allows the optical indication to be illuminated and detected at the second side 34 of the substrate 30.
  • FIG. 3 is a flow chart of an embodiment of a method of detecting hybridization of a molecule to a molecular receptor.
  • the method is utilized in conjunction with a molecular detection device as described herein. It is noted, however, that the method may also be utilized for detecting hybridization in other devices.
  • the method includes a step of providing a substrate which defines a binding site for receiving the molecular receptor.
  • the substrate can be provided within a molecular detection device, such as embodiments of the molecular detection device described herein.
  • the substrate is formed of a material, such as glass, which allows a transmission of light therethrough.
  • the method includes an optional step of generating an electric field using a transparent electrode proximate to the binding site.
  • the electric field is generated to attract molecules to the binding site to enhance hybridization to the molecular receptor.
  • a step of illuminating the binding site through the substrate is performed to produce an optical indication of a molecule which has hybridized to the molecular receptor.
  • a transparent electrode is included, the binding site and the optical indication are illuminated through the transparent electrode.
  • the binding site can be illuminated from a side of the substrate opposite to a side on which the binding site is defined.
  • the binding site can be illuminated by a light source, such as a light-emitting diode, fabricated into the substrate.
  • a step of detecting the optical indication through the substrate is performed. If a transparent electrode is included, the optical indication is detected through the transparent electrode.
  • the optical indication can be detected from a side of the substrate opposite to the side on which the binding site is defined.
  • the optical indication can be detected by an optical detector, such as a photodiode, fabricated on the substrate.
  • the method includes an optional step of generating an electric field using the transparent electrode to de-hybridize the molecule from the molecular receptor.
  • FIG. 4 is a schematic perspective view of a preferred embodiment of an integrated molecular detection chip in accordance with the present invention.
  • the integrated molecular detection chip includes a substrate 60 which defines a plurality of binding sites 62.
  • the plurality of binding sites 62 are formed by wells 64 etched on a top surface 66 of the substrate 60.
  • the plurality of selective binding sites 62 are arranged as a matrix in this embodiment. In alternative embodiments, the plurality of binding sites 62 can be arranged as another type of array.
  • FIG. 5 is a schematic cross-sectional view of the preferred embodiment of the integrated molecular detection chip.
  • Specific DNA receptors 70 having specific base pair sequences are deposited into the wells 64.
  • the specific DNA receptors 70 can be deposited using techniques which include, but are not limited to, robotic dispensing techniques and self-assembly techniques.
  • each of the plurality of binding sites 62 is a respective one of a plurality of optical detectors 72.
  • Each of the plurality of optical detectors 72 includes an a-Si photodiode fabricated into a bottom surface 74 of the substrate 60.
  • the plurality of optical detectors 72 can be matrix addressed using a-Si or poly-Si thin-film transistors to form an imaging array with a one-to-one correspondence to the binding sites 62.
  • Each of the plurality of light sources 76 includes a polymer light-emitting diode fabricated into the bottom surface 74 of the substrate 60.
  • the plurality of light sources 76 have a spectral output corresponding to absorption bands of fluorophores used to provide the optical indication.
  • a color filter 78 is incorporated over each of the plurality of optical detectors 72 to screen out light emanating from the plurality of light sources 76.
  • each of the plurality of binding sites 62 has a respective one of a plurality of transparent electrodes 80 integrated in proximity thereto.
  • the transparent electrodes 80 generate electric fields for field-enhanced hybridization and de-hybridization.
  • the transparent electrodes 80 are individually addressable for self- addressing and self-assembly purposes.
  • the integrated molecular detection chip can further include a polymer membrane at each of the plurality of binding sites 62 to aid in binding molecules thereto. In operation, appropriate DNA sequences hybridize onto selective ones of the plurality of binding sites 62 using either conventional, field-assisted, or thermally- assisted hybridization.
  • phase-sensitive detecting techniques can be incorporated by modulating a drive current to each of the plurality of optical detectors 72. These techniques are beneficial to enhance sensitivity of the detection.
  • the hybridized sites can then be detected to determine specific nucleotide sequences within the DNA sample.
  • the molecular detection chip can then be fully de- hybridized using chemical, thermal, or field-assisted means so that the hybridization and detection process can be repeated.
  • the various embodiments of the present invention integrate a binding site, a light source, and an optical detector on a single substrate, they provide a significant improvement in that a fully integrated molecular detection device is produced.
  • the various embodiments of the present invention as herein-described include a transparent electrode proximate to each binding site to provide field- assisted hybridization and de-hybridization.
  • the transparency of the electrode allows an optical indication of binding to be both illuminated and detected through the electrode.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Physics & Mathematics (AREA)
  • Pathology (AREA)
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  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Urology & Nephrology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
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  • Chemical Kinetics & Catalysis (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

Un substrat (12) définit un site de liaison (14) permettant d'accueillir un récepteur moléculaire (16). Une source lumineuse (18) éclaire le site de liaison (14) au travers du substrat (12) de façon à donner une indication optique d'une molécule (19) hybridée au récepteur moléculaire (16). Le détecteur optique (20) détecte l'indication optique au travers du substrat (12). De préférence, le substrat (12) définit un premier côté sur lequel est défini le site de liaison, et un second côté, opposé au premier côté, et sur lequel une source lumineuse et le détecteur optique sont intégrés. Il est possible d'inclure au niveau du site de liaison une électrode transparente en cas d'hybridation et de déshybridation assistées sur place.
PCT/US1997/005534 1996-04-17 1997-04-03 Dispositif optique integre de detection moleculaire et procede correspondant WO1997039144A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU24365/97A AU2436597A (en) 1996-04-17 1997-04-03 Integrated optical molecular detection device and method therefor

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US63407396A 1996-04-17 1996-04-17
US08/634,073 1996-04-17

Publications (1)

Publication Number Publication Date
WO1997039144A1 true WO1997039144A1 (fr) 1997-10-23

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PCT/US1997/005534 WO1997039144A1 (fr) 1996-04-17 1997-04-03 Dispositif optique integre de detection moleculaire et procede correspondant

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AU (1) AU2436597A (fr)
FR (1) FR2747786B1 (fr)
ID (1) ID19531A (fr)
IL (1) IL120626A0 (fr)
WO (1) WO1997039144A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004035834A1 (fr) * 2002-10-17 2004-04-29 Radius Biosciences, Inc. Procede mettant en oeuvre un substrat d'oxyde d'indium-etain dans un criblage a haut debit
US7125710B2 (en) * 2000-09-07 2006-10-24 Yokogawa Electric Corporation Apparatus for measuring the genetic sequence of biopolymers
WO2017199025A1 (fr) * 2016-05-17 2017-11-23 Cambridge Display Technology Limited Appareil d'excitation et de détection optiques de fluorescence
WO2017198709A1 (fr) * 2016-05-17 2017-11-23 Cambridge Display Technology Limited Dispositif d'excitation optique de fluorescence

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10015821A1 (de) * 2000-03-30 2001-10-18 Infineon Technologies Ag Sensor zum Erfassen von makromolekularen Biopolymeren, Sensoranordnung, Verfahren zum Erfassen von makromolekularen Biopolymeren und Verfahren zum Herstellen eines Sensors zum Erfassen von makromolekularen Biopolymeren
US20060241351A1 (en) * 2005-04-08 2006-10-26 Rosemount Analytical Inc. Integrated optical device for luminescence sensing

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Publication number Priority date Publication date Assignee Title
US4915812A (en) * 1986-06-20 1990-04-10 Molecular Devices Corporation Zero volume cell
US5527670A (en) * 1990-09-12 1996-06-18 Scientific Generics Limited Electrochemical denaturation of double-stranded nucleic acid
US5580794A (en) * 1993-08-24 1996-12-03 Metrika Laboratories, Inc. Disposable electronic assay device

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Publication number Priority date Publication date Assignee Title
DE3869237D1 (de) * 1987-07-07 1992-04-23 Siemens Ag Sensor fuer gase oder ionen.
GB8827853D0 (en) * 1988-11-29 1988-12-29 Ares Serono Res & Dev Ltd Sensor for optical assay
US5439647A (en) * 1994-02-25 1995-08-08 Fiberchem, Inc. Chip level waveguide sensor

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4915812A (en) * 1986-06-20 1990-04-10 Molecular Devices Corporation Zero volume cell
US5527670A (en) * 1990-09-12 1996-06-18 Scientific Generics Limited Electrochemical denaturation of double-stranded nucleic acid
US5580794A (en) * 1993-08-24 1996-12-03 Metrika Laboratories, Inc. Disposable electronic assay device

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7125710B2 (en) * 2000-09-07 2006-10-24 Yokogawa Electric Corporation Apparatus for measuring the genetic sequence of biopolymers
WO2004035834A1 (fr) * 2002-10-17 2004-04-29 Radius Biosciences, Inc. Procede mettant en oeuvre un substrat d'oxyde d'indium-etain dans un criblage a haut debit
WO2017199025A1 (fr) * 2016-05-17 2017-11-23 Cambridge Display Technology Limited Appareil d'excitation et de détection optiques de fluorescence
WO2017198709A1 (fr) * 2016-05-17 2017-11-23 Cambridge Display Technology Limited Dispositif d'excitation optique de fluorescence
GB2550560A (en) * 2016-05-17 2017-11-29 Sumitomo Chemical Co Device for optically exciting fluorescence
GB2550561A (en) * 2016-05-17 2017-11-29 Sumitomo Chemical Co Apparatus for optically exciting and detecting fluorescence
US20190162664A1 (en) * 2016-05-17 2019-05-30 Cambridge Display Technology Limited Device for optically exciting fluorescence
US20190293561A1 (en) * 2016-05-17 2019-09-26 Cambridge Display Technology Limited Apparatus for optically exciting and detecting fluorescence
US10989663B2 (en) 2016-05-17 2021-04-27 Cambridge Display Technology Limited Device for optically exciting fluorescence

Also Published As

Publication number Publication date
IL120626A0 (en) 1997-08-14
ID19531A (id) 1998-07-16
FR2747786A1 (fr) 1997-10-24
FR2747786B1 (fr) 2000-06-16
AU2436597A (en) 1997-11-07

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