Substrates, systems and methods for analyzing materials

• US 7,820,983 B2
• Filed: 08/31/2007
• Issued: 10/26/2010
• Est. Priority Date: 09/01/2006
• Status: Active Grant

• Current Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Original Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Inventors: Turner, Stephen, Lundquist, Paul
• Primary Examiner(s): Porta; David P
• Assistant Examiner(s): Kim; Kiho
• Application Number:
  US11/849,157
• Publication Number:
  US 20080128627A1
• Time in Patent Office: 1,152 Days
• Field of Search: 250/458.1
• US Class Current: 250/458.1
• CPC Class Codes: G01N 2021/7786 Fluorescence
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/648 using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
• CPC Class Codes: G01N 21/7703 using reagent-clad optical fibres or optical waveguides using measurement of total internal reflection or attenuated total reflection G01N21/552; optical fibres or waveguides per se G02B
• CPC Class Codes: G02B 6/43 Arrangements comprising a plurality of opto-electronic elements and associated optical interconnections

Abstract

Substrates, systems and methods for analyzing materials that include waveguide arrays disposed upon or within the substrate such that evanescent fields emanating from the waveguides illuminate materials disposed upon or proximal to the surface of the substrate, permitting analysis of such materials. The substrates, systems and methods are used in a variety of analytical operations, including, inter alia, nucleic acid analysis, including hybridization and sequencing analyzes, cellular analyzes and other molecular analyzes.

Claims

1. An analytical device, comprising:

a substrate comprising a first surface and at least a first optical waveguide disposed upon the first surface; and

an individual analyte disposed sufficiently proximal to the first surface and external to the waveguide to be illuminated by an evanescent field emanating from the waveguide when light is passed through the waveguide, wherein the individual analyte is immobilized on the first surface such that a signal from the individual analyte is optically resolvable from any other signal from any other analyte immobilized on the first surface and illuminated by the evanescent field.

2. The device of claim 1, wherein the substrate comprises a plurality of waveguides disposed upon the first surface that are optically coupled to a common originating waveguide disposed upon the first surface.

3. The device of claim 1, wherein the at least first waveguide is disposed within the first surface of the substrate.

4. The device of claim 1, wherein the substrate comprises at least 10 waveguides disposed upon the first surface.

5. The device of claim 1, wherein the substrate comprises at least 20 waveguides disposed upon the first surface.

6. The device of claim 1, wherein the substrate comprises at least 40 waveguides disposed upon the first surface.

7. The device of claim 1, wherein the substrate comprises SiO₂.

8. The device of claim 1, wherein the substrate comprises a polymer.

9. The device of claim 1, further comprising at least a first excitation radiation source optically coupled to the waveguide to pass excitation light therethrough.

10. The device of claim 9, wherein the at least first excitation radiation source is coupled to the first optical waveguide by at least a first optical fiber.

11. The device of claim 1, further comprising at least two excitation radiation sources optically coupled to the first optical waveguide to pass excitation light therethrough, each of the two excitation radiation sources providing light of a different spectrum.

12. The device of claim 11, wherein the first optical waveguide is further coupled to at least one optical device for controlling light passage through the first optical waveguide.

13. The device of claim 1, further comprising a mask layer disposed over the first surface of the substrate, the mask layer covering at least a first portion of the waveguide on the first surface and not covering at least a second portion of the waveguide on the first surface.

14. The device of claim 13, wherein the mask layer comprises a plurality of apertures disposed therethrough, the apertures providing access to the at least a portion of the waveguide on the first surface.

15. The device of claim 14, wherein one of said apertures provides structural confinement for the individual analyte.

16. The device of claim 15, wherein said one of said apertures is a zero mode waveguide.

17. The device of claim 13, wherein the mask layer and the substrate layer comprise the same material.

18. The device of claim 17, wherein the same material comprises a silica based material.

19. The device of claim 18, wherein the silica based material comprises SiO₂.

20. The device of claim 13, wherein the mask layer comprises a polymer.

21. The device of claim 13, wherein the mask layer comprises a metal.

22. The device of claim 13, wherein the mask layer blocks the evanescent field above portions of the first optical waveguide.

23. The device of claim 13, wherein the mask layer is aligned with an optical train.

24. The device of claim 1, wherein the first optical waveguide is imbedded beneath the first surface of the substrate.

25. The device of claim 1, wherein a plurality of analytes disposed sufficiently proximal to the first surface to be illuminated by the evanescent field are disposed in a pattern on the substrate, further wherein the plurality of analytes comprises said individual analyte and said any other analyte.

26. The device of claim 1, wherein the first optical waveguide has a cross sectional dimension between about 0.1 μ

m and about 10 μ

M.

27. The device of claim 1, wherein the individual analyte is immobilized on the first surface via interaction with an immobilized complex.

28. The device of claim 27, wherein the immobilized complex comprises an enzyme.

29. The device of claim 28, wherein the enzyme is a polymerase enzyme.