Compensator for multiple surface imaging

• US 8,071,962 B1
• Filed: 08/12/2011
• Issued: 12/06/2011
• Est. Priority Date: 05/05/2008
• Status: Active Grant

• Current Assignee: Illumina Incorporated
• Original Assignee: Illumina Incorporated
• Inventors: Feng, Wenyi, Bryant, Jason, Buermann, Dale
• Primary Examiner(s): Kim, Kiho
• Application Number:
  US13/209,306
• Publication Number:
  US 20110301044A1
• Time in Patent Office: 116 Days
• Field of Search: 250/458.1, 250/459.1
• US Class Current: 250/459.1
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: G01N 2021/0346 Capillary cells; Microcells
• CPC Class Codes: G01N 2021/6419 Excitation at two or more wavelengths
• CPC Class Codes: G01N 2021/6421 Measuring at two or more wavelengths
• CPC Class Codes: G01N 21/05 Flow-through cuvettes G01N21/09 takes precedence; handling fluid samples G01N1/10
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G02B 21/0024 Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
• CPC Class Codes: G02B 2207/113 Fluorescence

Abstract

A system and method for imaging biological samples on multiple surfaces of a support structure are disclosed. The support structure may be a flow cell through which a reagent fluid is allowed to flow and interact with the biological samples. Excitation radiation from at least one radiation source may be used to excite the biological samples on multiple surfaces. In this manner, fluorescent emission radiation may be generated from the biological samples and subsequently captured and detected by detection optics and at least one detector. The detected fluorescent emission radiation may then be used to generate image data. This imaging of multiple surfaces may be accomplished either sequentially or simultaneously. In addition, the techniques of the present invention may be used with any type of imaging system. For instance, both epifluorescent and total internal reflection methods may benefit from the techniques of the present invention.

Claims

1. A method for detecting a biological sample, comprising:

(a) detecting radiation emitted from a first emissive component of a biological sample disposed on a first surface of a multi-surface support structure using a detector;

(b) inserting corrective optics between the detector and the multi-surface support structure; and

(c) detecting radiation emitted from a second emissive component of a biological sample disposed on a second surface of the multi-surface support structure using the detector and the corrective optics, wherein the first and second surfaces are in an arrangement whereby one of the surfaces is disposed between the detector and the other surface, wherein the corrective optics reduce aberration of detection at one of the surfaces due to the arrangement.

2. The method of claim 1, wherein the first surface is disposed between the detector and the second surface.

3. The method of claim 2, wherein a fluid is present between the two surfaces and the corrective optics reduce aberration due to the fluid.

4. The method of claim 3, wherein the detector is initially configured for diffraction-limited detection on the second surface and the corrective optics are configured for diffraction-limited detection on the first surface.

5. The method of claim 3, wherein the aberration comprises spherical aberration.

6. The method of claim 1, wherein the first surface is disposed between a source of radiation and the second surface.

7. The method of claim 1, wherein step (b) comprises inserting a corrective device between an objective and the multi-surface support structure, wherein the objective is disposed between the detector and the multi-surface support structure.

8. The method of claim 1, wherein step (b) comprises inserting a corrective lens between the detector and an objective, wherein the objective is disposed between the detector and the multi-surface support structure.

9. The method of claim 1, wherein steps (a) and (c) comprise obtaining an image of the surfaces.

10. The method of claim 1, wherein steps (a) and (c) comprise focusing a line of radiation to an area on the first or second surface comprising the first or second emissive component.

11. The method of claim 1, wherein the emitted radiation is detected using epifluorescent excitation.

12. The method of claim 1, wherein excitation radiation is directed towards the first and second surfaces by total internal reflection excitation.

13. The method of claim 1, wherein the first and second emissive components comprise fluorescently tagged nucleic acids of a nucleic acid sample.

14. The method of claim 1, wherein the first and second emissive components comprise separate features in arrays of features on the first and second surfaces, respectively.

15. The method of claim 1, wherein the multi-surface support structure comprises a flow cell.

16. The method of claim 15, wherein the flow cell is mounted on a detecting station.

17. The method of claim 16, further comprising:

(d) repeating steps (a) through (c) while maintaining the flow cell on the detecting station.

18. The method of claim 17, wherein the repeating of steps (a) through (c) comprises treating the flow cell with several repeated cycles of a nucleic acid sequencing process.

19. The method of claim 17, wherein the repeating of steps (a) through (c) comprises repeated steps of delivering one or more reagents to the first and second emissive components of the biological sample disposed on the first and second surfaces.

20. A system for detecting radiation on a multi-surface support structure, comprising:

a detecting station configured to position a multi-surface support structure in a focus plane of an objective;

an optical train comprising the objective, detecting optics configured to focus the optical train on a first surface of the multi-surface support structure via the objective, and corrective optics configured to focus the optical train on a second surface of the multi-surface support structure and configured to reduce aberration of detection at the first or second surface;

a radiation source configured to direct excitation radiation towards the first and second surfaces;

detection optics configured to capture emitted radiation returned from the first and second surfaces via the optical train; and

a detector positioned to detect the captured radiation.

21. The system of claim 20, wherein the corrective optics comprise a corrective device configured to be removably inserted between the objective and the multi-surface support structure.

22. The system of claim 20, wherein the corrective optics comprise a corrective lens configured to be removably inserted between the detector and the objective.

23. The system of claim 20, wherein the radiation source is configured to direct the excitation radiation toward the first and second surfaces at several different wavelengths, and the detection optics and detector are configured to capture and detect the emitted radiation returned in response to each wavelength.

24. The system of claim 20, wherein the multi-surface support structure is present at the detecting station.

25. The system of claim 24, wherein first and second emissive components of a biological sample are disposed on the first and second surfaces, respectively.

26. The system of claim 25, wherein the multi-surface support structure includes a fluid in an interior volume and in contact with the first and second emissive components of the biological sample.

27. The system of claim 20, wherein the first surface is disposed between the radiation source and the second surface.