Multi-site detection apparatus

• US 5,891,630 A
• Filed: 04/15/1996
• Issued: 04/06/1999
• Est. Priority Date: 11/19/1991
• Status: Expired due to Term

• Current Assignee: Daniel J. Ehrlich, John Shumaker, Kenneth Loren Beattie, Mark Hollis, Michael E. Hogan, Mitchell D. Eggers
• Original Assignee: Daniel J. Ehrlich, John Shumaker, Kenneth Loren Beattie, Mark Hollis, Michael E. Hogan, Mitchell D. Eggers
• Inventors: Hogan, Michael E., Hollis, Mark, Eggers, Mitchell D., Shumaker, John, Ehrlich, Daniel J., Beattie, Kenneth Loren
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US08/633,861
• Time in Patent Office: 1,086 Days
• Field of Search: 204/400, 204/403, 204/153.1, 204/153.12, 422/82.01, 422/82.02, 435/6, 435/7.1, 435/7.2, 435/287.1, 435/287.2, 436/518, 436/524, 436/525, 436/527, 436/806, 436/809
• US Class Current: 506/3
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00511 Walls of reactor vessels
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00653 the compounds being bound to electrodes embedded in or on the solid supports
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• CPC Class Codes: B01J 2219/00711 Light-directed synthesis
• CPC Class Codes: B01J 2219/00713 Electrochemical synthesis
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C12Q 1/6825 : Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/6832 : Enhancement of hybridisation reaction
• CPC Class Codes: C12Q 1/6837 : using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 1/6874 : involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 2565/607 : being a sensor, e.g. electrode
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 21/253 : for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 27/221 : by investigating the dielectric properties using microwaves G01N22/00; measuring loss factors or dielectric constants per se G01R27/26
• CPC Class Codes: G01N 27/226 : Construction of measuring vessels; Electrodes therefor
• CPC Class Codes: G01N 33/54373 : involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 33/5438 : Electrodes
• CPC Class Codes: Y10S 436/806 : Electrical property or magnetic property
• CPC Class Codes: Y10S 436/809 : Multifield plates or multicontainer arrays

Abstract

A method and apparatus for identifying molecular structures within a sample substance using an array having a plurality of test sites upon which the sample substance is applied. Each test site includes a probe formed therein to bond with an associated target molecular structure. An electrical signal is applied to the test site and the electrical properties of the test sites are detected to determine which probes have bonded to an associated target molecular structure.

Claims

1. Apparatus for identifying molecular structures within a sample substance, comprising:

a plurality of test sites formed in a single substrate, said plurality of test sites having probes attached thereto which bind to a specific target molecular structure, such that different ones of said plurality of test sites have different probes for binding to different specific target molecular structures;

a plurality of electrodes associated with said plurality of test sits, each test site having at least a first electrode attached thereto for electrical coupling with a second electrode, wherein said first electrode and said second electrode are disposed to form a capacitor in conjunction with the sample substance;

circuitry, coupled to said plurality of electrodes, for applying an electronic signal to said plurality of test sites at multiple frequencies; and

circuitry for detecting a frequency dependent electrical property of the sample substance at said plurality of test sites to determine which probes have bound to an associated target molecular structure.

2. The apparatus of claim 1 wherein one or more of said probes binds to a DNA molecule.

3. The apparatus of claim 1 wherein one or more of said probes binds to an RNA molecule.

4. The apparatus of claim 1 wherein one or more of said probes include oligonucleotide probes.

5. The apparatus of claim 1 wherein one or more of said probes binds to a cell.

6. The apparatus of claim 1 wherein one or more of said probes binds to an antibody.

7. The apparatus of claim 6 wherein one or mote of said probes include anti-antibody probes.

8. The apparatus of claim 1 wherein one or more of said probes include peptide probes.

9. The apparatus of claim 1 wherein said first electrode includes a first capacitor plate formed in each test site and said second electrode includes a capacitor plate common to each test site.

10. The apparatus of claim 1 wherein said first electrode includes a first capacitor plate formed in each test site and said second electrode includes a second capacitor plate formed in each test site.

11. The apparatus of claim 1 wherein said circuitry for applying an electronic signal to said plurality of test sites at multiple frequencies includes circuitry for applying an electronic signal to said plurality of test sites over a predetermined range of frequencies.

12. The apparatus of claim 11 wherein said detecting circuitry includes circuitry for detecting said frequency dependent electrical property of the sample substance at said plurality of test sites within said predetermined range of frequencies.

13. The apparatus of claim 12 wherein said detecting circuitry further includes circuitry for comparing said detected frequency dependent electrical property to a known value of said frequency dependent electrical property at a test site with unbonded probes.

14. An integrated circuit for identifying molecular structures within a sample substance, comprising:

a plurality of test sites formed in a single substrate;

a plurality of electrodes associated with said plurality of test sites, each test site having at least a first electrode attached thereto for electrical coupling with a second electrode such that the sample substance acts as a dielectric between said first and second electrodes;

probes attached at respective test sites, said probes being of various structures for binding with respective predetermined molecular structures; and

circuitry, coupled to said plurality of electrodes, for detecting a frequency dependent electrical property of the sample substance at said plurality of test sites to determine whether probes associated with each test sites have bound with the respective predetermined molecular structures.

15. Apparatus for identifying molecular structures within a sample substance, comprising:

a plurality of test sites formed in a single substrate, said plurality of test sites having respective probes attached thereto which bind to a specific target molecular structure, such that different test sites have probes for binding to different specific target molecular structures;

a plurality of electrodes associated with said plurality of test sites, each one of said plurality of test sites having at least one electrode formed therein for forming a capacitive coupling with a second electrode;

circuitry, coupled to said electrodes, for applying an electronic signal to said plurality of test sites over a range of frequencies; and

circuitry for detecting capacitive parameters of the sample substance at said plurality of test sites within said range of frequencies to determine a relaxation frequency and comparing said relaxation frequency to a known frequency at a test site with unbound probes to determine which probes have bound to an associated target molecular structure.

16. Apparatus for identifying molecular structures within a sample substance, comprising:

a plurality of test sites formed as respective wells in a single substrate, said test sites having respective probes attached to said wells to specifically bind to an associated target molecular structure, such that different test sites have probes for binding to different target molecular structures;

a plurality of electrodes formed on sidewalls of said wells;

a ground plate common to each well for providing a capacitive coupling to said plurality of electrodes;

circuitry, coupled to said electrodes, for applying an electronic signal to said test sites over a range of frequencies; and

circuitry for detecting a frequency dependent electrical property of the sample substance at said test sites to determine which probes have bonded to an associated target molecular structure.

17. Apparatus for identifying molecular structures within a sample substance, comprising:

a plurality of test sites formed in a single substrate, said test sites attached to respective probes for binding to a target molecular structure, such that different test sites have probes which bind to different target molecular structures;

a plurality of electrodes associated with each of said test sites, each test site having a least one electrode formed therein for electrical coupling with a second electrode;

circuitry, coupled to said electrodes, for applying an electronic signal to said test sites at multiple frequencies; and

circuitry for detecting a frequency dependent property at said test sites by measuring capacitive properties of said test sites to determine which ones of said probes have bound to an associated target molecular structure.

18. An integrated circuit for identifying molecular structures within a sample substance, comprising:

a plurality of test sites formed in a single substrate;

a plurality of electrodes associated with each of said test sites, each of said test sites having at least one electrode formed therein for electrical coupling with a second electrode;

a plurality of probes attached at respective ones of said test sites, said probes being of various structures to bind with respective predetermined molecular structures; and

circuitry, coupled to said electrodes, for detecting frequency dependent capacitive properties at said test sites at multiple frequencies to determine whether the probes associated with the test sites have bound with the respective predetermined molecular structures.

19. Apparatus for identifying molecular structures within a sample substance, comprising:

a plurality of test sites formed in a single substrate, said test sites having probe means which bind to a target molecular structure, such that different test sites have probe means which bind to different target molecular structures;

electrode means associated with said plurality of test sites, such that said electrode means form a capacitor in conjunction with the sample substance at said test sites;

signal means for applying an electronic signal to said test sites at multiple frequencies; and

detecting means for detecting a frequency dependent electrical property at said test sites to determine which probes have bound to an associated target molecular structure.

20. Apparatus for identifying molecular structures within a sample substance, comprising.

a plurality of test sites formed in a single substrate, said test sites having respective probes attached thereto which bind to a target molecular structure, such that different test sites have probes which bind to different target molecular structures;

a plurality of electrodes associated with each of said sites, each of said test sites having at least a first electrode attached thereto for electrical coupling with a second electrode, wherein the first electrode and the second electrode are disposed to form a capacitor in conjunction with the sample substance;

circuitry, coupled to said plurality electrodes, for applying an electronic signal to test sites at multiple frequencies; and

circuitry for detecting frequency dependent electrical properties at said test sites to determine which probes have bound to an associated target molecular structure, wherein the frequency dependent electrical properties include at least relaxation frequency, permittivity and dispersion.