Optical detection system for flow cytometry

• US 6,549,275 B1
• Filed: 08/02/2000
• Issued: 04/15/2003
• Est. Priority Date: 08/02/2000
• Status: Expired due to Term

• Current Assignee: Honeywell International Inc.
• Original Assignee: Honeywell International Inc.
• Inventors: Marta, Teresa M., Cox, James Allen, Padmanabhan, Aravind, Zook, J. David, Cabuz, Cleopatra
• Primary Examiner(s): Stafira, Michael P.
• Application Number:
  US09/630,927
• Time in Patent Office: 986 Days
• Field of Search: 356/39, 356/317, 356/411, 250/461.2, 250/358.2, 250/226
• US Class Current: 356/39
• CPC Class Codes: G01F 1/7086 using optical detecting arrangements
• CPC Class Codes: G01N 15/1434 Optical arrangements
• CPC Class Codes: G01N 15/1456 without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
• CPC Class Codes: G01N 2015/011 with lysing, e.g. of erythrocytes
• CPC Class Codes: G01N 2015/012 Red blood cells
• CPC Class Codes: G01N 2015/016 White blood cells
• CPC Class Codes: G01N 2015/0238 Single particle scatter
• CPC Class Codes: G01N 2015/1027 Determining speed or velocity of a particle
• CPC Class Codes: G01N 35/085 Flow Injection Analysis

Abstract

An optical detection system for flow cytometry that uses two or more light sources positioned laterally at different distances from the central axis of the flow stream for providing light through different parts of the flow stream. By using two or more light sources, the particle position can be detected, and the alignment and width of the core stream can be monitored and controlled. In addition, the velocity and size of the particles can be more accurately determined than when only a single light source is used.

Claims

1. An optical detection system for analyzing predetermined characteristics of a flow stream, the flow stream having a central axis along the direction of flow, the optical detection system comprising:

two or more light sources positioned on a common axis at an angle relative to the central axis of the flow stream for providing light through different parts of the flow stream;

light receiving means for receiving the light from the two or more light sources, and for providing at least one signal in response thereto; and

processing means for receiving the at least one signal from the light receiving means and for using the at least one signal for analyzing the predetermined characteristics of the flow scream.

2. An optical detection system according to claim 1, wherein the two or more light sources are positioned along a light source axis that is offset relative to the central axis of flow of the flow stream.

3. An optical detection system according to claim 1, wherein the two or more spaced light sources are part of an array of VCSEL devices.

4. An optical detection system according to claim 3, wherein the VCSEL devices operate in the red spectrum.

5. An optical detection system according to claim 1, wherein each of the two or more light sources have a corresponding lens.

6. An optical detection system according to claim 5, wherein each lens is an integrated micro lens.

7. An optical detection system according to claim 5, wherein selected lenses focus the light at a central plane that includes the central axis of the flow stream.

8. An optical detection system according to claim 7, wherein the light that is focused at the central plane is used for detecting the small angle scattering (SALS) produced by one or more particles in the flow stream.

9. An optical detection system according to claim 8, wherein the one or more particles include white blood cells.

10. An optical detection system according to claim 9, wherein the white blood cells include neutrophils and/or lymphocytes white blood cells.

11. An optical detection system according to claim 7, wherein the light that is focused at the central plane is used for determining the velocity of one or more particles in the flow stream.

12. An optical detection system according to claim 7, wherein the light that is focused at the central plane is used for determining the size of one or more particles in the flow stream.

13. An optical detection system according to claim 5, wherein selected lenses provide substantially collimated light through the flow stream.

14. An optical detection system according to claim 13, wherein the substantially collimated light is used for detecting the forward angle scattering (FALS) produced by one or more particles in the flow stream.

15. An optical detection system according to claim 14, wherein the one or more particles include white blood cells.

16. An optical detection system according to claim 15, wherein the white blood cells include neutrophils and/or lymphocytes white blood cells.

17. An optical detection system according to claim 1, wherein the two or more light sources include a first set of light sources positioned along a first light source axis that is rotated relative to the central axis of the flow stream, light from the first set of light sources is focused at a central plane that includes the central axis of the flow stream.

18. A method for determining the alignment of one or more particles in a flow stream relative to a central axis of the flow stream, the method comprising:

activating two or more light sources positioned laterally across the central axis of the flow stream to provide light through different parts of the flow stream;

monitoring an output response of two or more detectors that receive the light from the two or more light sources;

detecting a change in the response of one or more of the detectors when a particle passes between the detector and the corresponding light source; and

determining the alignment of the one or more particles in the flow stream relative to the central axis of the flow stream by noting which detector or detectors produced a change in response.

19. A method for determining the velocity of one or more particles in a flow stream, the flow stream having a width and a length, the length extending from an upstream position to a downstream position, the method comprising the steps of:

activating at least one upstream light source at the upstream position of the flow stream to provide light through the flow stream;

monitoring an output response of at least one upstream detector that receives the light from the upstream light source;

activating at least one downstream light source at a downstream position of the flow stream, a downstream light source positioned at a different position along the width of the flow stream than an upstream light source to provide light through a different part of the flow stream;

monitoring an output response of at least one downstream detector that receives the light from the downstream light source;

detecting a change in the response of the at least one upstream detector when a particle passes between the at least one upstream detector and the at least one upstream light source;

detecting a change in the response of the at least one downstream detector when the same particle passes between the at least one downstream detector and the at least one downstream light source; and

determining a time lag between the change in response of the at least one upstream detector and the change in response of the at least one downstream detector, the time lag being related to the speed of the particle.

20. A method for determining the small angle scattering (SALS) produced by one or more particles in a flow stream, the method comprising:

activating two or more light sources positioned on a common axis at an angle relative to the flow stream to provide light through different parts of the flow stream;

focusing the light provided by the two or more light sources at the flow stream;

receiving the light from the two or more light sources using at least one detector, each detector having a detector region laterally spaced from an in-line position of the corresponding light source; and

determining the SALS produced by the one or more particles by examining the response of the at least one detector.

21. A method according to claim 20 wherein each detector has at least two detector regions for each of the light sources, one positioned in one direction relative to an in-line position of the corresponding light source and another positioned in another direction relative to the in-line position of the corresponding light source.

22. A method for determining the forward angle scattering (FALS) produced by one or more particles in a flow stream, the method comprising:

activating two or more light sources laterally positioned across the flow stream to provide light through the flow stream;

collimating the light provided by the two or more light sources at the flow stream;

receiving the light from the two or more light sources using at least one detector, each detector having a detector region laterally spaced from an in-line position of the corresponding light source; and

determining the FALS produced by the one or more particles by examining the response of the at least one detector.

23. A method according to claim 22, wherein each of the at least one detectors is annular in shape.

24. An optical detection system for analyzing predetermined characteristics of a flow stream, the flow stream having a central axis along the direction of flow, the optical detection system comprising:

a linear array of light sources for providing light through different parts of the flow stream, the linear array of light sources oriented at an angle with respect to the central axis of the flow stream;

a linear array of light detectors for receiving the light from the linear array of light sources, and for providing at least one signal in response thereto; and

processing means for receiving the at least one signal from the linear array of light detectors and for using the at least one signal for analyzing the predetermined characteristics of the flow stream.

25. An optical detection system for analyzing predetermined characteristics of a flow stream, the flow stream having a central axis along the direction of flow, the optical detection system comprising:

two or more light sources positioned laterally across the central axis of the flow stream for providing light through different parts of the flow stream;

light receiving means for receiving the light from the two or more light sources, and for providing at least one signal in response thereto; and

processing means for receiving the at least one signal from the light receiving means and for using the at least one signal for analyzing the predetermined characteristics of the flow stream; and

wherein the two or more light sources are positioned along a light source axis that is offset relative to the central axis of flow of the flow stream.

26. An optical detection system according to claim 25, wherein the two or more light sources collectively provide substantially constant light intensity across the flow stream.

27. An optical detection system for analyzing predetermined characteristics of a flow stream, the flow stream having a central axis along the direction of flow, the optical detection system comprising:

two or more light sources positioned laterally across the central axis of the flow stream for providing light through different parts of the flow stream;

light receiving means for receiving the light from the two or more light sources, and for providing at least one signal in response thereto; and

processing means for receiving the at least one signal from the light receiving means and for using the at least one signal for analyzing the predetermined characteristics of the flow stream; and

wherein the two or more light sources include a first set of light sources positioned along a first light source axis that is rotated relative to the central axis of the flow stream, light from the first set of light sources is focused at a central plane that includes the central axis of the flow stream.

28. An optical detection system according to claim 27, wherein the light receiving means includes a first set of light detectors for receiving light from the first set of light sources after the light passes through the flow stream, each of the first set of light detectors having a corresponding lens that focuses the light substantially on the corresponding light detector.

29. An optical detection system according to claim 28, wherein the first set of light sources and the first set of light detectors are used to detect the alignment of the flow of one or more particles relative to the central axis of the flow stream.

30. An optical detection system according to claim 28, wherein the first set of light sources and the first set of light detectors are used to detect the speed of one or more particles in the flow stream.

31. An optical detection system according to claim 28, wherein the first set of light sources and the first set of light detectors are used to detect the size of one or more particles in the flow stream.

32. An optical detection system according to claim 27, wherein the two or more light sources include a second set of light sources positioned along a second light source axis that is rotated relative to the central axis of the flow stream and offset from the first light source axis, each of the second set of light sources having a corresponding lens that focuses the light at a central plane that includes the central axis of the flow stream.

33. An optical detection system according to claim 32, wherein the light receiving means includes a second set of light detectors, each of the second set of light detectors having a detector region laterally spaced from an in-line position of the corresponding light source.

34. An optical detection system according to claim 33, wherein each of the second set of light detectors has at least two detector regions for each of the second set of light sources, one positioned in one direction relative to an in-line position of the corresponding light source and another positioned in another direction relative to the in-line position of the corresponding light source, each of the second set of light detectors having a corresponding lens that focuses the light substantially on the corresponding light detector.

35. An optical detection system according to claim 34, wherein the second set of light detectors are used to detect the small angle scattering (SALS) produced by one or more particles in the flow stream.

36. An optical detection system according to claim 34, wherein the second set of light detectors include a third detector in-line with the corresponding light source, the third detector used in conjunction with the first set of detectors to detect the speed of one or more particles in the flow stream.

37. An optical detection system according to claim 33, wherein the two or more light sources include a third set of light sources positioned along a third light source axis that is rotated relative to the central axis of the flow stream, each of the third set of light sources having a corresponding lens that provides substantially collimated light through the flow stream.

38. An optical detection system according to claim 37, wherein the light receiving means includes a third set of light detectors positioned in line with the third set of light sources, each of the third set of light detectors having a corresponding lens that focuses the substantially collimated light substantially on the corresponding light detector.

39. An optical detection system according to claim 38, wherein each of the third set of light detectors are annular in shape.

40. An optical detection system according to claim 39, wherein the third set of light sources and the third set of annular light detectors are used to detect the forward angle scattering (FALS) produced by the selected particles in the flow stream.

41. A method for determining the small angle scattering (SALS) produced by one or more particles in a flow stream, the method comprising:

activating two or more light sources positioned laterally across the flow stream to provide light through different parts of the flow stream;

focusing the light provided by the two or more light sources at the flow stream;

receiving the light from the two or more light sources using at least one detector, each detector having a detector region laterally spaced from an in-line position of the corresponding light source; and

determining the SALS produced by the one or more particles by examining the response of the at least one detector; and

wherein each detector has at least two detector regions for each of the light sources, one positioned in one direction relative to an in-line position of the corresponding light source and another positioned in another direction relative to the in-line position of the corresponding light source.

42. An optical detection system for analyzing predetermined characteristics of a flow stream, the flow stream having a central axis along the direction of flow, the optical detection system comprising:

two or more light sources positioned laterally across the central axis of the flow stream for providing light through different parts of the flow stream;

light receiving means for receiving the light from the two or more light sources, and for providing at least one signal in response thereto; and

processing means for receiving the at least one signal from the light receiving means and for using the at least one signal for analyzing the predetermined characteristics of the flow stream; and

wherein the two or more light sources, having overlapping Gaussian beams, collectively provide substantially constant light intensity across the flow stream.

43. An optical detection system for analyzing predetermined characteristics of a flow stream, the flow stream having a central axis along the direction of flow, the optical detection system comprising:

two or more light sources positioned laterally across the central axis of the flow stream for providing light through different parts of the flow stream;

light receiving means for receiving the light from the two or more light sources, and for providing at least one signal in response thereto; and

processing means for receiving the at least one signal from the light receiving means and for using the at least one signal for analyzing the predetermined characteristics of the flow stream; and

wherein the two or more light sources are positioned along an axis that is offset relative to the central axis of flow of the flow stream.