Optical instrument including excitation source

• US 20040038390A1
• Filed: 05/19/2003
• Published: 02/26/2004
• Est. Priority Date: 05/17/1999
• Status: Active Grant

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Inventors: Oldham, Mark F., Young, Eugene F., Boege, Steven J.
• Granted Patent: US 7,387,891 B2
• Time in Patent Office: Days
• US Class Current: 435/288.7
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• 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 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 2021/6441 with two or more labels
• CPC Class Codes: G01N 2021/6463 Optics
• CPC Class Codes: G01N 2021/6482 Sample cells, cuvettes
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 2201/0628 Organic LED [OLED]
• CPC Class Codes: G01N 2201/0635 Structured illumination, e.g. with grating
• CPC Class Codes: Y10S 435/808 Optical sensing apparatus

Abstract

An optical instrument is provided for simultaneously illuminating two or more spaced-apart reaction regions with an excitation beam generated by a light source. A collimating lens can be disposed along a beam path between the light source and the reaction regions to form bundles of collimated excitation beams, wherein each bundle corresponds to a respective reaction region. Methods of analysis using the optical instrument are also provided.

Claims

1. An instrument comprising:

a block with a plurality of spaced-apart reaction regions;

a light emitting diode source adapted to direct excitation beams toward the plurality of reaction regions; and

a collimating lens system disposed along a path of excitation beams between the light emitting diode source and the plurality of reaction regions, wherein the collimating lens system is capable of collimating excitation beams emitted from the light emitting diode source into two or more spaced-apart bundles of collimated excitation beams, and wherein the instrument is capable of directing each of two or more bundles of collimated excitation beams toward a respective reaction region of the plurality of reaction regions.

2. The instrument of claim 1, further comprising:

a focusing lens disposed along a path of at least one of the two or more bundles of collimated excitation beams between the collimating lens system and the plurality of reaction regions.

3. The instrument of claim 2, wherein the focusing lens is disposed adjacent the reaction region.

4. The instrument of claim 2, wherein the focusing lens is a Fresnel lens.

5. The instrument of claim 1, wherein the collimating lens system includes a collimating lens and a mask.

6. The instrument of claim 1, wherein a sample is disposed in at least one of the reaction regions and the sample includes a dye that is capable of emitting an emission beam when illuminated with a respective one of two or more bundles of collimated excitation beams.

7. The instrument of claim 6, wherein the sample comprises components for nucleic acid sequence amplification.

8. The instrument of claim 7, wherein the sample comprises components for polymerase chain reaction.

9. The instrument of claim 1, wherein the plurality of reaction regions comprises 96 reaction regions.

10. The instrument of claim 1, wherein the collimating lens is disposed about one focal length of the collimating lens away from the light emitting diode source.

11. The instrument of claim 1, wherein the light emitting diode source comprises a light emitting diode having a wattage of greater than about one microwatt.

12. The instrument of claim 1, wherein the light emitting diode has a wattage of about 5 microwatts or greater.

13. The instrument of claim 1, further comprising an excitation filter disposed along an excitation beam path between the light emitting diode source and the plurality of reaction regions.

14. The instrument of claim 13, wherein the excitation filter comprises a long pass filter, a bandpass filter, a multiple bandpass filter, or a combination thereof.

15. The instrument of claim 1, wherein there is a correspondence of 1 to at least 4 between the light emitting diode source and the bundles of collimated excitation beams, respectively.

16. The instrument of claim 1, wherein the collimating lens system comprises a Fresnel lens.

17. The instrument of claim 1, wherein the collimating lens system comprises a molded glass sphere.

18. The instrument of claim 1, further comprising a mask disposed between the collimating lens system and the plurality of reaction regions.

19. The instrument of claim 18, wherein the mask is optically opaque.

20. The instrument of claim 18, wherein the mask comprises anodized aluminum.

21. The instrument of claim 18, wherein the mask is in contact with the collimating lens system.

22. The instrument of claim 20, wherein the mask is adjacent a focusing lens, and the focusing lens is disposed along a beam path of two or more bundles of collimated excitation beams between the collimating lens system and the plurality of reaction regions.

23. The instrument of claim 1, further comprising at least one field lens disposed along an excitation beam path between the collimating lens system and the plurality of reaction regions.

24. The instrument of claim 23, wherein the at least one field lens comprises a Fresnel lens.

25. The instrument of claim 23, comprising two field lenses spaced-apart by the sum of the focal lengths of the two field lenses.

26. The instrument of claim 1, further comprising a mirror disposed along an excitation beam path between the collimating lens system and the plurality of reaction regions.

27. The instrument of claim 1, further comprising a transition filter disposed along an excitation beam path between the collimating lens system and the plurality of reaction regions.

28. The instrument of claim 27, wherein the transition filter comprises a long pass filter, a bandpass filter, a multiple bandpass filter, or a combination thereof.

29. The instrument of claim 1, further comprising a detector disposed to receive emission beams emitted from each of the plurality of reaction regions.

30. The instrument of claim 29, wherein the detector comprises a camera, a charge-coupled detector, a photodiode, a photomultiplier, a CMOS, a CID, or a combination thereof.

31. The instrument of claim 29, wherein the detector is capable of generating a first data set, representative of detected emission beams, and wherein the instrument further comprises a processor capable of receiving a first data set from the detector and processing the first data set.

32. The instrument of claim 1, wherein the light emitting diode source includes an organic light emitting diode.

33. An instrument comprising:

a block with a plurality of spaced-apart reaction regions;

a solid state laser source adapted to direct excitation beams toward the plurality of reaction regions; and

a collimating lens system disposed along a path of excitation beams between the solid state laser source and the plurality of reaction regions, wherein the collimating lens system is capable of collimating excitation beams emitted from the solid state laser source into two or more spaced-apart bundles of collimated excitation beams, and wherein the instrument is capable of directing each of two or more bundles of collimated excitation beams toward a respective reaction region of the plurality of reaction regions.

34. The instrument of claim 33, further comprising:

a focusing lens disposed along a path of at least one of the two or more bundles of collimated excitation beams between the collimating lens system and the plurality of reaction regions.

35. The instrument of claim 34, wherein the focusing lens is disposed adjacent the reaction region.

36. The instrument of claim 34, wherein the focusing lens is a Fresnel lens.

37. An instrument comprising:

a block with a plurality of spaced-apart reaction regions;

a micro-wire laser source adapted to direct excitation beams toward the plurality of reaction regions; and

a collimating lens system disposed along a path of excitation beams between the micro-wire laser source and the plurality of reaction regions, wherein the collimating lens system is capable of collimating excitation beams emitted from the micro-wire laser source into two or more spaced-apart bundles of collimated excitation beams, and wherein the instrument is capable of directing each of two or more bundles of collimated excitation beams toward a respective reaction region of the plurality of reaction regions.

38. The instrument of claim 37, further comprising:

a focusing lens disposed along a path of at least one of the two or more bundles of collimated excitation beams between the collimating lens system and the plurality of reaction regions.

39. The instrument of claim 38, wherein the focusing lens is disposed adjacent the reaction region.

40. The instrument of claim 38, wherein the focusing lens is a Fresnel lens.

41. A method of illuminating a plurality of spaced-apart reaction regions with excitation beams, the method comprising:

providing an instrument comprising a light emitting diode source, a collimating lens system, and a block including the plurality of spaced-apart reaction regions, at least one of the reaction regions comprising a sample;

generating excitation beams with the light emitting diode source;

passing the excitation beams through the collimating lens system to form two or more spaced-apart bundles of collimated excitation beams; and

focusing each of the two or more spaced-apart bundles of collimated excitation beams into respective ones of the plurality of spaced-apart reaction regions.

42. The method of claim 41, further comprising detecting emission beams emitted from the plurality of spaced-apart reaction regions, with a detector.

43. The method of claim 42, further comprising generating a first dataset representative of emission beams detected by the detector.

44. The method of claim 41, further comprising processing the first data set with a processor.

45. The method of claim 42, further comprising passing emission beams emitted from the plurality of spaced-apart reaction regions through an emission filter.

46. The method of claim 41, further comprising passing the two or more spaced-apart bundles of collimated excitation beams from the collimating lens system through a first field lens, and from the first field lens through a second field lens.

47. The method of claim 41, wherein the step of focusing each of the two or more spaced-apart bundles of collimated excitation beams comprises passing each of the two or more spaced-apart bundles of collimated excitation beams through a focusing lens.

48. The method of claim 47, wherein each focusing lens is a reaction region lens disposed adjacent a respective reaction region.

49. The method of claim 47, wherein the focusing lens is a Fresnel lens.

50. The method of claim 41, further comprising removing extraneous excitation beams with a mask.

51. The method of claim 50, wherein the mask is disposed along an excitation beam path between the collimating lens system and the plurality of reaction regions.

52. The method of claim 41, wherein the sample comprises components for a nucleic acid sequence amplication reaction.

53. The method of claim 52, wherein the nucleic acid sequence amplication reaction comprises a polymerase chain reaction.

54. A method of illuminating a plurality of spaced-apart reaction regions with excitation beams, the method comprising:

providing an instrument comprising a solid state laser source, a collimating lens system, and a block including the plurality of spaced-apart reaction regions, at least one of the reaction regions comprising a sample;

generating excitation beams with the solid state laser source;

passing the excitation beams through the collimating lens system to form two or more spaced-apart bundles of collimated excitation beams; and

focusing each of the two or more spaced-apart bundles of collimated excitation beams into respective ones of the plurality of spaced-apart reaction regions.

55. The method of claim 54, wherein the step of focusing each of the two or more spaced-apart bundles of collimated excitation beams comprises passing each of the two or more spaced-apart bundles of collimated excitation beams through a focusing lens.

56. The method of claim 55, wherein each focusing lens is a reaction region lens disposed adjacent a respective reaction region.

57. The method of claim 55, wherein the focusing lens is a Fresnel lens.

58. A method of illuminating a plurality of spaced-apart reaction regions with excitation beams, the method comprising:

providing an instrument comprising a micro-wire laser source, a collimating lens system, and a block including the plurality of spaced-apart reaction regions, at least one of the reaction regions comprising a sample;

generating excitation beams with the micro-wire laser source;

passing the excitation beams through the collimating lens system to form two or more spaced-apart bundles of collimated excitation beams; and

focusing each of the two or more spaced-apart bundles of collimated excitation beams into respective ones of the plurality of spaced-apart reaction regions.

59. The method of claim 58, wherein the step of focusing each of the two or more spaced-apart bundles of collimated excitation beams comprises passing each of the two or more spaced-apart bundles of collimated excitation beams through a focusing lens.

60. The method of claim 59, wherein each focusing lens is a reaction region lens disposed adjacent a respective reaction region.

61. The method of claim 59, wherein the focusing lens is a Fresnel lens.