Integrated illumination of optical analytical devices

• US 9,624,540 B2
• Filed: 02/21/2014
• Issued: 04/18/2017
• Est. Priority Date: 02/22/2013
• Status: Active Grant

• Current Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Original Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Inventors: Lundquist, Paul, Turner, Stephen
• Primary Examiner(s): WHITE, DENNIS MICHAEL
• Application Number:
  US14/187,198
• Publication Number:
  US 20140287964A1
• Time in Patent Office: 1,152 Days
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: G01N 2021/7789 Cavity or resonator
• 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 21/6454 using an integrated detector array
• CPC Class Codes: G01N 21/648 using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
• CPC Class Codes: G01N 21/7746 the waveguide coupled to a cavity resonator
• CPC Class Codes: G01N 2201/08 Optical fibres; light guides
• CPC Class Codes: G02B 2006/12147 Coupler
• CPC Class Codes: G02B 6/0229 characterised by nanostructures, i.e. structures of size less than 100 nm, e.g. quantum dots

Abstract

Optical analytical devices and their methods of use are provided. The devices are useful in the analysis of highly multiplexed optical reactions in large numbers at high densities, including biochemical reactions, such as nucleic acid sequencing reactions. The devices include integrated illumination elements and optical waveguides for illumination of the optical reactions. The devices further provide for the efficient coupling of optical excitation energy from the waveguides to the optical reactions. Optical signals emitted from the reactions can thus be measured with high sensitivity and discrimination using features such as spectra, amplitude, and time resolution, or combinations thereof. The devices of the invention are well suited for miniaturization and high throughput.

Claims

1. An analytical device comprising:

a substrate;

an integrated illumination element comprising a waveguide source;

a plurality of illumination volumes; and

a plurality of detector elements;

wherein

the integrated illumination element is disposed above the surface of the substrate;

the plurality of detector elements are disposed below the surface of the substrate;

the integrated illumination element is partly surrounded by an opaque layer; and

the plurality of illumination volumes are contained in a plurality of nanowells disposed above the surface of the substrate and in the same layer of the device as the integrated illumination element, wherein the nanowells are configured in a row adjacent to the integrated illumination element, wherein at least a first nanowell comprises a bottom surface and a first side surface, and wherein the first nanowell is optically coupled to the integrated illumination element through an opening in the opaque layer and to one of the detector elements.

2. The analytical device of claim 1, wherein the first nanowell is optically coupled to the integrated illumination element through the first side surface of the first nanowell.

3. The analytical device of claim 2, wherein the opaque layer partly covers the first side surface of the first nanowell.

4. The analytical device of claim 2, wherein the opaque layer does not cover the first side surface of the first nanowell.

5. The analytical device of claim 2, wherein the first nanowell further comprises a second side surface, wherein the second side surface comprises a micromirror.

6. The analytical device of claim 5, wherein the micromirror increases the optical coupling of an illumination volume contained in the first nanowell to the integrated illumination element.

7. The analytical device of claim 5, wherein the second side surface has a concave shape.

8. The analytical device of claim 5, wherein the second side surface is angled toward the bottom surface of the first nanowell.

9. The analytical device of claim 1, wherein the first nanowell is optically coupled to the integrated illumination element through the bottom surface of the first nanowell.

10. The analytical device of claim 9, wherein the opaque layer completely covers the first side surface of the first nanowell.

11. The analytical device of claim 9, wherein the first nanowell is optically coupled to the integrated illumination element through a transfer waveguide disposed in the substrate.

12. The analytical device of claim 11, wherein the transfer waveguide illuminates no more than one of the nanowells.

13. The analytical device of claim 11, wherein the transfer waveguide illuminates more than one of the nanowells.

14. The analytical device of claim 1, wherein the first nanowell is optically coupled to the detector element through the bottom surface of the first nanowell.

15. The analytical device of claim 1, wherein the opaque layer partly surrounding the integrated illumination element comprises a micromirror.

16. The analytical device of claim 15, wherein the micromirror increases the optical coupling of an illumination volume contained in the first nanowell to the integrated illumination element.

17. The analytical device of claim 15, wherein the opaque layer partly surrounding the integrated illumination element is cylindrical.

18. The analytical device of claim 1, further comprising a filter layer disposed between the surface of the substrate and the plurality of detector elements.

19. The analytical device of claim 18, wherein the filter layer decreases the transmission of optical energy from the integrated illumination element to the plurality of detector elements.

20. The analytical device of claim 1, wherein at least one detector element of the plurality of detector elements further comprises a microlens.

21. The analytical device of claim 1, wherein at least one detector element of the plurality of detector elements further comprises a light redirection cone.

22. The analytical device of claim 1, further comprising an analyte disposed within at least one illumination volume.

23. The analytical device of claim 22, wherein the analyte comprises a biological sample.

24. The analytical device of claim 23, wherein the biological sample comprises a nucleic acid.

25. The analytical device of claim 23, wherein the biological sample comprises a polymerase enzyme.

26. The analytical device of claim 1, wherein the analytical device comprises at least 1,000, at least 10,000, at least 100,000, at least 1,000,000, or at least 10,000,000 illumination volumes.

27. The analytical device of claim 1, wherein at least one detector element of the plurality of detector elements further comprises a spectral diversion element.

28. The analytical device of claim 1, wherein at least one detector element of the plurality of detector elements further comprises a light redirection cone.

29. The analytical device of claim 1, wherein the integrated illumination element comprises an array of two or more waveguides.

30. The analytical device of claim 29, wherein the two or more waveguides are two or more channel waveguides.

31. The analytical device of claim 1, wherein the row of nanowells comprises at least 100 nanowells.

32. The analytical device of claim 1, wherein the integrated illumination element comprises an array of two or more channel waveguides, and wherein each of the two or more waveguides is optically coupled to a row of at least 100 nanowells.