Apparatus and methods for analyzing samples
View Patent ↗The present invention relates to apparatus, systems, and methods for analyzing biological samples. The apparatus, systems, and methods can involve using a vacuum source to pull microfluidic volumes through analytical equipment, such as flow cells and the like. Additionally, the invention involves using optical equipment in conjunction with the analytical equipment to analyze samples and control the operation thereof.
1. A system for analyzing a sample, comprising:
a) a flow cell comprising a sample, wherein said sample comprises: i) nucleic acid templates, ii) primer sequences, iii) fluorescently labeled nucleotides, and iv) a polymerase;
b) a computer module, wherein said computer module comprises a processor;
c) an optical instrument for viewing said sample in said flow cell, wherein said optical instrument comprises an image capture device configured to:
i) capture an image of said sample, and
ii) send said image to said computer module for analysis; and
d) a lighting system for illuminating said sample in said flow cell, wherein said lighting system comprises:
i) one or more analytical light sources for illuminating said fluorescently labeled nucleotides in said sample; and
ii) a focusing light source configured to focus said optical instrument on said sample.
2. The system of claim 1 , further comprising: e) a moveable stage, wherein said flow cell is located on said moveable stage.
3. The system of claim 2 , wherein said moveable stage is moveable in the X, Y, and/or Z directions relative to said optical instrument.
4. The system of claim 1 , wherein said optical instrument is moveable in the X, Y, and/or Z directions.
5. The system of claim 1 , wherein said lighting system further comprises a translator configured to modify the angle of an optical path from said one or more analytical light sources.
6. The system of claim 1 , wherein said image capture device comprises a camera.
7. The system of claim 1 , wherein said image capture device comprises a charge coupled device (CCD).
8. The system of claim 1 , said flow cell further comprises a surface, and wherein said nucleic acid templates are linked to said surface via a nucleic acid sequence.
9. The system of claim 1 , wherein said optical instrument further comprises a microscope objective.
10. The system of claim 1 , further comprising an emission filter.
11. The system of claim 10 , wherein said emission filter is configured to aid in detecting an emission spectrum from said fluorescently labeled nucleotides.
12. The system of claim 1 , wherein said one or more analytical light sources comprises a red laser.
13. The system of claim 1 , wherein said one or more analytical light sources comprises a red laser and a green laser.
14. The system of claim 1 , wherein said focusing light source comprises a laser.
15. A method for sequencing comprising:
a) adding reagents to a flow cell, wherein said flow cell comprises an entry port, an exit port, a surface, a primer, and a nucleic acid template, wherein said nucleic acid template is linked to said surface and hybridized to said primer, wherein said reagents comprise fluorescently labeled nucleotides and a polymerase, and wherein said reagents are added to said entry port of said flow cell;
b) detecting an incorporated fluorescently labeled nucleotide added to said primer by said polymerase, wherein said detecting comprises:
i) illuminating said flow cell with a lighting system, wherein said lighting system comprises:
A) one or more analytical light sources for illuminating said incorporated fluorescently labeled nucleotide; and
B) a focusing light source configured to focus an optical instrument on said flow cell; and
ii) capturing an image of said incorporated fluorescently labeled nucleotide with said optical instrument; and
c) sending said image to a computer module for analysis such that the identity of said incorporated fluorescently labeled nucleotide is determined.
16. The method of claim 15 , wherein said flow cell is located on a moveable stage.
17. The method of claim 16 , further comprising moving said moveable stage to as to position said flow cell relative to said optical instrument.
18. The method of claim 15 , wherein said detecting further comprises moving said optical instrument with respect to said flow cell.
19. The method of claim 15 , wherein steps a)-c) are repeated at least once.
20. The method of claim 15 , wherein steps a)-c) are repeated until the complete sequence of said nucleic acid template is determined.
21. The method of claim 15 , wherein said nucleic acid template is linked to said surface via a nucleic acid sequence.
22. The method of claim 15 , wherein said lighting system further comprises a translator configured to modify the angle of an optical path from said one or more analytical light sources.
23. The method of claim 15 , wherein said image capture device comprises a camera.
24. The method of claim 15 , wherein said image capture device comprises a charge coupled device (CCD).
25. The method of claim 15 , wherein said optical instrument further comprises a microscope objective.
26. The method of claim 15 , wherein said image is captured by said optical instrument through an emission filter.
27. The method of claim 15 , wherein said one or more analytical light sources comprises a red laser.
28. The method of claim 15 , wherein said one or more analytical light sources comprises a red laser and a green laser.
29. The method of claim 15 , wherein said focusing light source comprises a laser.
30. The method of claim 15 , further comprising a step of removing reagents from said flow cell via said exit port.
31. A system for analyzing a sample, comprising:
a flow cell comprising a sample, wherein said sample comprises: i) nucleic acid templates, ii) primer sequences, iii) fluorescently labeled nucleotides, and iv) a polymerase;
a lighting system for illuminating the sample in the flow cell;
a computer module, wherein said computer module comprises a processor; and
an optical instrument for viewing the sample in the flow cell, wherein said optical instrument comprises an image capture device configured to: i) capture an image of said sample, and ii) send said image to said computer module for analysis;
wherein the lighting system comprises:
one or more analytical light sources, each light source defining an optical path that intersects the sample; and
a focusing light source operating with any one of the analytical light sources to focus said optical instrument on the sample.
32. The system of claim 31 , further comprising a moveable stage, wherein said flow cell is located on said moveable stage.
33. The system of claim 32 , wherein said moveable stage is moveable in the X, Y, and/or Z directions relative to said optical instrument.
34. The system of claim 31 , wherein said optical instrument is moveable in the X, Y, and/or Z directions.
35. The system of claim 31 , wherein said lighting system further comprises a translator configured to modify the angle of said optical path from said one or more analytical light sources.
36. The system of claim 31 , wherein said image capture device comprises a camera.
37. The system of claim 31 , wherein said image capture device comprises a charge coupled device (CCD).
38. The system of claim 31 , wherein said flow cell further comprises a surface, and wherein said nucleic acid template sequences are linked to said surface via a nucleic acid sequence.
39. The system of claim 31 , wherein said optical instrument further comprises a microscope objective.
40. The system of claim 31 , further comprising an emission filter.
41. The system of claim 38 , wherein said emission filter is configured to aid in detecting an emission spectrum from said fluorescently labeled nucleotides.
42. The system of claim 31 , wherein said one or more analytical light sources comprises a red laser.
43. The system of claim 31 , wherein said one or more analytical light sources comprises a red laser and a green laser.
44. The system of claim 31 , wherein said focusing light source comprises a laser.
45. A method for sequencing comprising:
a) adding reagents to a flow cell, wherein said flow cell comprises an entry port, an exit port, a surface, a primer, and a nucleic acid template, wherein said nucleic acid template is linked to said surface and hybridized to said primer, wherein said reagents comprise fluorescently labeled nucleotides and a polymerase, and wherein said reagents are added to said entry port of said flow cell;
b) detecting an incorporated fluorescently labeled nucleotide added to said primer by said polymerase, wherein said detecting comprises:
i) illuminating said flow cell with a lighting system, wherein said lighting system comprises:
A) one or more analytical light sources, each light source defining an optical path that intersects said flow cell; and
B) a focusing light source operating with any one of said analytical light sources to focus an optical instrument on said flow cell; and
ii) capturing an image of said incorporated fluorescently labeled nucleotide with said optical instrument; and
c) sending said image to a computer module for analysis such that the identity of said incorporated fluorescently labeled nucleotide is determined.
46. The method of claim 45 , wherein said flow cell is located on a moveable stage.
47. The method of claim 46 , further comprising moving said moveable stage to as to position said flow cell relative to said optical instrument.
48. The method of claim 45 , wherein said detecting further comprises moving said optical instrument with respect to said flow cell.
49. The method of claim 45 , wherein steps a)-c) are repeated at least once.
50. The method of claim 45 , wherein steps a)-c) are repeated until the complete sequence of said nucleic acid template is determined.
51. The method of claim 45 , wherein said nucleic acid template is linked to said surface via a nucleic acid sequence.
52. The method of claim 45 , wherein said lighting system further comprises a translator configured to modify the angle of said optical path from said one or more analytical light sources.
53. The method of claim 45 , wherein said image capture device comprises a camera.
54. The method of claim 45 , wherein said image capture device comprises a charge coupled device (CCD).
55. The method of claim 45 , wherein said optical instrument further comprises a microscope objective.
56. The method of claim 45 , wherein said image is captured by said optical instrument through an emission filter.
57. The method of claim 45 , wherein said one or more analytical light sources comprises a red laser.
58. The method of claim 45 , wherein said one or more analytical light sources comprises a red laser and a green laser.
59. The method of claim 45 , wherein said focusing light source comprises a laser.
60. The method of claim 45 , further comprising the step of removing reagents from said flow cell via said exit port.