Substrates, systems and methods for analyzing materials

• US 7,834,329 B2
• Filed: 10/31/2007
• Issued: 11/16/2010
• 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:
  US11/981,795
• Publication Number:
  US 20080152281A1
• Time in Patent Office: 1,112 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 analyses, cellular analyses and other molecular analyses.

Claims

1. A method of detecting a signal from an analyte, comprising:

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

providing 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;

directing light through the first waveguide such that the evanescent field from the waveguide illuminates the individual analyte; and

detecting a signal from the individual analyte.

2. The method of claim 1, wherein the individual analyte is confined proximal to a selected portion of the first surface.

3. The method of claim 2, wherein a structural barrier confines the individual analyte proximal to the selected portion of the first surface.

4. The method of claim 3, wherein the structural barrier comprises a well in a mask layer, and further wherein the mask layer is an element of the substrate and disposed over the first surface, and further wherein the well provides confinement of the individual analyte proximal to a portion of the first surface.

5. The method of claim 4, wherein the mask layer blocks the evanescent field above portions of the first optical waveguide.

6. The method of claim 5, wherein said detecting is performed using an optical train and further wherein the optical train is aligned with the mask layer.

7. The method of claim 1, wherein the individual analyte comprises a fluorescent moiety, and the step of directing light comprises directing light having an excitation wavelength for the fluorescent moiety.

8. The method of claim 1, wherein the individual analyte is confined proximal to a portion of the first surface by a hydrophobic coating on the first surface.

9. The method of claim 8, wherein the hydrophobic coating surrounds the portion of the first surface.

10. The method of claim 1, wherein the substrate comprises at least a first fluidic channel disposed over and in fluid communication with the first surface, and the providing step comprises delivering the individual analyte proximal to the first surface through the first fluidic channel.

11. The method of claim 1, wherein the individual analyte comprises a complex of a polymerase enzyme, a target nucleic acid sequence, a primer sequence complementary to at least a portion of the target sequence, and a nucleotide or nucleotide analog.

12. The method of claim 1, wherein the first optical waveguide is coupled to multiple light sources, wherein the multiple light sources have different characteristics.

13. The method of claim 12, wherein the different characteristics are different wavelengths.

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

15. A method of detecting a signal from an individual analyte, comprising:

providing a substrate comprising a first surface and at least a first optical waveguide disposed upon the first surface, wherein the substrate has a refractive index that is lower than that of the first optical waveguide and further wherein the first optical waveguide extends into and is at least partially enclosed by the substrate;

providing 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;

directing light through the first waveguide such that the evanescent field from the waveguide illuminates the individual analyte; and

detecting a signal from the individual analyte.