Compensator for multiple surface imaging
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.
1. A method for detecting features of an array, comprising:
(a) providing a support structure comprising a first interior surface and a second interior surface that are in a generally mutually facing relation, wherein the first interior surface and the second interior surface are separated by a fluid passage, and wherein an array of features is distributed on the first interior surface and on the second interior surface, wherein the features are present at a density of at least 1,000 sites per square millimeter;
(b) detecting the array at a resolution to distinguish the features on the first interior surface; and
(c) detecting the array at a resolution to distinguish the features on the second interior surface.
2. The method of claim 1 , wherein the features are distributed in a spatially ordered pattern on the first interior surface and on the second interior surface.
3. The method of claim 1 , wherein the features are distributed in a random spatial distribution on the first interior surface and on the second interior surface.
4. The method of claim 1 , wherein (b) and (c) further comprise translating the support structure, whereby the detecting occurs at successive regions of the first interior surface and at successive regions of the second interior surface, respectively.
5. The method of claim 1 , wherein the features comprise fluorescent labels, and wherein the detecting comprises detecting the fluorescent labels.
6. The method of claim 5 , wherein the first interior surface and the second interior surface are detected from the same side of the support structure in (b) and (c).
7. The method of claim 6 , wherein the first interior surface is disposed between an excitation source and the second interior surface during the detecting of (c).
8. The method of claim 7 , wherein the first interior surface and the second interior surface are excited by total internal reflection.
9. The method of claim 6 , wherein the first interior surface is disposed between a detector and the second interior surface during the detecting of (c).
10. The method of claim 9 , further comprising inserting or removing corrective optics between the detector and the support structure after (b) and before (c).
11. The method of claim 9 , further comprising inserting or removing corrective optics between the detector and the support structure after (b) and before (c).
12. The method of claim 1 , wherein the detecting in (b) and (c) produce images of the first interior surface and the second interior surface.
13. The method of claim 12 , wherein the images have a resolution of 10 microns or less.
14. The method of claim 1 , wherein the array of features comprises a plurality of nucleic acid sites that is distributed on the first interior surface and on the second interior surface.
15. The method of claim 14 , further comprising:
(d) repeating (b) and (c) in a process of sequencing the nucleic acids at the nucleic acid sites.
16. The method of claim 14 , further comprising:
(d) introducing a fluorescent reagent to the support structure to add fluorescent tags to the nucleic acid sites.
17. The method of claim 16 , further comprising:
(d) repeating (b), (c) and (d) in a process of sequencing the nucleic acids at the nucleic acid sites.
18. A method for detecting features of an array, comprising:
(a) providing a support structure comprising a first interior surface and a second interior surface that are in a generally mutually facing relation, wherein the first interior surface and the second interior surface are separated by a fluid passage, and wherein an array of features is distributed on the first interior surface and on the second interior surface;
(b) detecting the array at a resolution between 0.1 and 50 microns to distinguish the features on the first interior surface; and
(c) detecting the array at a resolution between 0.1 and 50 microns to distinguish the features on the second interior surface.
19. The method of claim 18 , wherein the features are distributed in a spatially ordered pattern on the first interior surface and on the second interior surface.
20. The method of claim 18 , wherein the features are distributed in a random spatial distribution on the first interior surface and on the second interior surface.
21. The method of claim 18 , wherein (b) and (c) further comprise translating the support structure, whereby the detecting occurs at successive regions of the first interior surface and at successive regions of the second interior surface, respectively.
22. The method of claim 18 , wherein the features comprise fluorescent labels, and wherein the detecting comprises detecting the fluorescent labels.
23. The method of claim 22 , wherein the first interior surface and the second interior surface are detected from the same side of the support structure in (b) and (c).
24. The method of claim 23 , wherein the first interior surface is disposed between an excitation source and the second interior surface during the detecting of (c).
25. The method of claim 24 , wherein the first interior surface and the second interior surface are excited by total internal reflection.
26. The method of claim 23 , wherein the first interior surface is disposed between a detector and the second interior surface during the detecting of (c).
27. The method of claim 18 , wherein the detecting in (b) and (c) produce images of the first interior surface and the second interior surface.
28. The method of claim 18 , wherein the array of features comprises a plurality of nucleic acid sites that is distributed on the first interior surface and on the second interior surface.
29. The method of claim 28 , further comprising:
(d) repeating (b) and (c) in a process of sequencing the nucleic acids at the nucleic acid sites.
30. The method of claim 28 , further comprising:
(d) introducing a fluorescent reagent to the support structure to add fluorescent tags to the nucleic acid sites.
31. The method of claim 30 , further comprising:
(d) repeating (b), (c) and (d) in a process of sequencing the nucleic acids at the nucleic acid sites.