Apparatus and methods for analyzing samples

• US 20060012784A1
• Filed: 11/16/2004
• Published: 01/19/2006
• Est. Priority Date: 07/19/2004
• Status: Active Grant

• Current Assignee: Fluidigm Corporation
• Original Assignee: Helicos Biosciences Corporation
• Inventors: Ulmer, Kevin
• Granted Patent: US 7,276,720 B2
• Time in Patent Office: Days
• US Class Current: 356/246
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2400/049 vacuum
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502738 characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: G01N 21/55 Specular reflectivity

Abstract

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.

Claims

1. A system for analyzing a sample, comprising:

a flow cell;

a passive vacuum source for pulling a volume through the flow cell;

a lighting system for illuminating the sample in the flow cell; and

an optical instrument for viewing the sample in the flow cell.

2. The system of claim 1, wherein the sample is predisposed within the flow cell.

3. The system of claim 2, wherein the volume reacts with the sample in the flow cell.

4. The system of claim 1, wherein the volume comprises the sample.

5. The system of claim 1, further comprising a stage for receiving the flow cell, wherein the stage is movable in at least one direction.

6. The system of claim 5, wherein the stage is movable in at least two orthogonal directions.

7. The system of claim 1, further comprising an image capture device for capturing an image of the sample.

8. The system of claim 7, wherein the image capture device comprises at least one of a charge-coupled device, a complementary metal oxide semiconductor device, a charge injection device, and a video camera.

9. The system of claim 1, wherein the lighting system comprises:

a first light source for analyzing the sample, light from the first light source defining a first optical path that intersects the sample; and

a second light source operating with the first light source for determining a position of the first optical path.

10. The system of claim 9, wherein the first light source and the second light source operate simultaneously.

11. The system of claim 9, wherein light from the second light source defines a second optical path at least partially coaxial with the first optical path.

12. The system of claim 9, wherein the second light source is directed to a position sensor for sensing an angle of incidence of the first optical path relative to the sample, and the relative orientation of the first optical path and the sample can be adjusted to vary the angle of incidence in response to a signal from the position sensor.

13. The system of claim 12, wherein the relative orientation of the first optical path and the sample can be adjusted to obtain substantially total internal reflection of the first light source at the sample.

14. The system of claim 9, wherein the first light source comprises a wavelength from about 390 nm to about 780 nm.

15. The system of claim 9, wherein the second light source comprises infrared light.

16. The system of claim 9, wherein at least one of the first light source and the second light source is selected from the group consisting of a laser, a light emitting diode, and a lamp.

17. The system of claim 7, further comprising a processor in electrical communication with the image capture device for collecting and processing data generated by the system.

18. The system of claim 17, further comprising storage for storing the data.

19. The system of claim 17, further comprising means for displaying at least one of the sample and the data.

20. The system of claim 1, wherein the system is used for single molecule detection.

21. The system of claim 20, wherein the flow cell includes a surface for receiving a nucleotide.

22. The system of claim 21, wherein the surface of the flow cell includes a primer for binding the nucleotide thereto.

23. The system of claim 1, wherein the flow cell comprises a channel for pulling the microfluidic volume therethrough.

24. The system of claim 1, wherein the passive vacuum source comprises:

a pump;

a pump driver; and

a reservoir, wherein the pump is connected to the reservoir to evacuate the reservoir, thereby creating a vacuum pressure therein.

25. The system of claim 24, wherein a ratio of a volume of the reservoir and the microfluidic volume is between about 1,000:

1 and about 2,000,000;

1.

26. The system of claim 25, wherein the ratio is about 200,000:

1.

27. The system of claim 1, further comprising a valve disposed between the vacuum source and the flow cell, wherein the valve comprises an open position to connect the flow cell to the vacuum source and a closed position to isolate the flow cell from the vacuum source.

28. The system of claim 24, further comprising a vacuum pressure indicator connected to the reservoir.

29. The system of claim 24, wherein the vacuum pressure is from about 1″

Hg to about 29″

Hg.

30. The system of claim 24, wherein the vacuum pressure is adjustable to vary a speed at which the volume passes through the flow cell.

31. A method of detecting single molecules comprising:

providing a flow cell that defines a channel that is treated to identify specific molecules;

applying a vacuum to the channel to pull a sample through the channel, the sample comprising single molecules; and

viewing the sample in the channel to identify the single molecules.

32. The method of claim 31, wherein the sample comprises a microfluidic volume comprising nucleotides.

33. The method of claim 32, wherein the flow cell comprises at least one of a slide and a coverslip treated to bind with a specific nucleotide.

34. The method of claim 31, wherein the step of viewing the flow cell comprises illuminating the flow cell with a lighting system.

35. The method of claim 34, wherein the step of illuminating the flow cell comprises:

shining light from a first light source along a first optical path that intersects the sample; and

shining a light from a second light source operating with the first light source so as to determine a position of the first optical path.

36. The method of claim 31, wherein the step of viewing the flow cell comprises using an image capture device.

37. The method of claim 31, wherein a processor is used to control the operation of the method.

38. The method of claim 37, further comprising the step of using a processor for collecting and processing data generated during the method.

39. The method of claim 38, further comprising the step of displaying at least one of the flow cell and the data.

40. The method of claim 31, wherein the step of applying a vacuum comprises exposing the flow cell to a passive vacuum source.

41. An apparatus for handling microfluidic volumes, the apparatus comprising:

a vacuum source; and

a flow cell in communication with the vacuum source, wherein the microfluidic volume is pulled through the flow cell by the passive vacuum source.