Multi-site detection apparatus

• US 5,532,128 A
• Filed: 12/12/1994
• Issued: 07/02/1996
• Est. Priority Date: 11/19/1991
• Status: Expired due to Term

• Current Assignee: Houston Advanced Research Center
• Original Assignee: Houston Advanced Research Center
• Inventors: Hollis, Mark, Hogan, Michael E., Beattie, Kenneth L., Eggers, Mitchell D., Shumaker, John, Ehrlich, Daniel J.
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): Chin, Christopher L.
• Application Number:
  US08/353,957
• Time in Patent Office: 568 Days
• Field of Search: 204/153.1, 204/153.12, 204/400, 204/403, 422/82.01, 422/82.02, 435/6, 435/7.1, 435/7.2, 435/287, 435/291, 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 test sites having respective probes attached thereto which specifically bind to a target molecular structure, such that different test sites have probes which specifically bind to different target molecular structures;

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

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

circuitry for detecting one of a plurality of frequency dependent electrical properties of the test sites to determine which probes have bound to an associated target molecular structure by measuring a dielectric property over a range of frequencies, wherein the plurality of frequency dependent electrical properties comprises permittivity, a dielectric relaxation frequency, and a dispersion of the test sites.

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

3. The apparatus of claim 2 wherein said one or more probes comprise oligonucleotide probes.

4. The apparatus of claim 1 wherein one or more of said probes bind to a RNA molecule.

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

6. The apparatus of claim 5 wherein said one or more probes comprise antibody probes.

7. The apparatus of claim 1 wherein one or more of said probes bind to an antibody.

8. The apparatus of claim 7 wherein said one or more probes comprise anti-antibody probes.

9. The apparatus of claim 8 wherein said one or more probes comprise peptide probes.

10. The apparatus of claim 1 wherein said test sites comprise:

a plurality of wells formed in the substrate, said wells defined by side and bottom walls;

probes for binding with a desired molecular structure formed in each well;

wherein said electrodes are disposed adjacent to said walls to form a capacitor in conjunction with the sample substance.

11. The apparatus of claim 10 wherein said capacitors each comprise a plate formed in each well which is capacitively coupled to a ground plate common to each well.

12. The apparatus of claim 11 wherein said capacitors each comprise first and second plates formed on the sidewalls of the wells.

13. The apparatus of claim 1 wherein said circuitry for applying an electronic signal comprises circuitry for applying a signal over a predetermined range of frequency.

14. The apparatus of claim 13 wherein said detecting circuitry comprises circuitry for measuring capacitance to determine said relaxation frequency within said range.

15. The apparatus of claim 14 wherein said detecting circuitry further comprises circuitry for comparing said relaxation frequency to a known frequency for a test site with unbound probes.

16. The apparatus of claim 1 wherein said circuitry for applying an electronic signal comprises circuitry for applying a signal at discrete predetermined frequencies at a selected test site.

17. The apparatus of claim 16 wherein said detecting circuitry comprises circuitry for measuring capacitance of the sample substance at said selected test site at each of said discrete predetermined frequencies to determine said dispersion at the predetermined frequencies and comparing the dispersion between said frequencies to determine whether binding with a target molecular structure has occurred at said selected test site.

18. The apparatus of claim 1 and further comprising circuitry for determining a structure based on the test sites having bonded probes.

19. The apparatus of claim 1 wherein said test sites are formed on a semiconductor surface.

20. 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 the test sites, each test site having at least one electrode attached thereto for electrical coupling with a second electrode, wherein the one electrode and the second electrode are disposed adjacent to said test sites to form a capacitor in conjunction with the sample substance;

probes attached to respective test sites, said probes being of various structures in order to specifically bind with respective predetermined molecular structures; and

circuitry, coupled to said electrodes, for measuring capacitive parameters at multiple frequencies detect one of a plurality of frequency dependent electrical properties of the test sites to determine whether the probes associated with the test sites have bound with the respective predetermined molecular structures, wherein the plurality of frequency dependent electrical properties of the test sites comprises permittivity a dielectric relaxation frequency, and a dispersion of the tests sites.

21. The integrated circuit of claim 20 and further comprising processing circuitry for processing information from said measuring circuitry.

22. The integrated circuit of claim 20 wherein each of said test sites comprise first and second conducting plates formed with a well, said probes being disposed between said conductive surfaces.

23. The integrated circuit of claim 22 wherein said first and second plates are parallel.

24. The integrated circuit of claim 22 wherein said measuring circuitry is operable to generate a signal over a range of frequencies to said plates of each of said test sites and measure capacitance responsive thereto to determine the dispersion over said range of frequencies.

25. The integrated circuit of claim 22 wherein said detecting circuitry is operable to generate a signal at discrete frequencies to said plates of each of said test sites and measure capacitance responsive thereto to determine dispersion at said discrete frequencies.

26. 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 specifically bind to a target molecular structure, such that different test sites have probes which specifically bind to different target molecular structures;

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

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

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

27. 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 which specifically bind 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 electrodes;

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

circuitry for detecting one of a plurality of frequency dependent electrical properties of the test sites by measuring capacitive parameters and dissipation of the test sites at multiple frequencies to determine which probes have bonded to an associated target molecular structure, wherein the plurality of frequency dependent electrical properties comprises permittivity, a dielectric relaxation frequency, and a dispersion of the test sites,

28. 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 which specifically bind to a target molecular structure, such that different test sites have probes which specifically bind to different target molecular structures;

a plurality of electrodes associated with the test sites, each test site having a least one electrode formed therein for electrical coupling with a second electrode, wherein the one electrode and the second electrode are disposed adjacent to said test sites to form a capacitor in conjunction with the sample substance;

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

circuitry for detecting a frequency dependent capacitive property of the test sites by measuring capacitive parameters of the test sites at multiple frequencies to determine which probes have bound to an associated target molecular structure, wherein the frequency despondent capacitive property of the tests sites is permittivity.

29. 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 the test sites, each test site having at least one electrode formed therein for electrical coupling with a second electrode, wherein the one electrode and the second electrode are disposed adjacent to said test sites to form a capacitor in conjunction with the sample substance;

probes attached to respective test sites, said probes being of various structures in order to specifically bind with respective predetermined molecular structures; and

circuitry, coupled to said electrodes, for detecting a frequency dependent capacitive property of the test sites by measuring capacitive parameters at multiple frequencies to determine whether the probes associated with the test sites have bound with the respective predetermined molecular structures, wherein the frequency dependent capacitive property of the tests sites is a dielectric relaxation frequency of the test sites.

30. 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 the test sites, each test site having at least one electrode formed therein for electrical coupling with a second electrode, wherein the one electrode and the second electrode are disposed adjacent to said test sites to form a capacitor in conjunction with the sample substance;

probes attached to respective test sites, said probes being of various structures in order to specifically bind with respective predetermined molecular structures; and

circuitry, coupled to said electrodes, for detecting a frequency dependent capacitive property of the test sites by measuring capacitive parameters at multiple frequencies to determine whether the probes associated with the test sites have bound with the respective predetermined molecular structures, wherein the frequency dependent capacitive property of the tests sites is a dispersion of the test sites.