Portable flow cytometry

• US 6,597,438 B1
• Filed: 08/02/2000
• Issued: 07/22/2003
• Est. Priority Date: 08/02/2000
• Status: Expired due to Term

• Current Assignee: Honeywell International Inc.
• Original Assignee: Honeywell International Inc.
• Inventors: Cabuz, Eugen Loan, Ohnstein, Thomas Raymond, Satren, Ernest Allen, Bonne, Ulrich, Cox, James Allen, Padmanabhan, Aravind, Zook, J. David, Cabuz, Cleopatra, Marta, Teresa M.
• Primary Examiner(s): Rosenberger, Richard A.
• Assistant Examiner(s): Barth, Vincent P.
• Application Number:
  US09/630,924
• Time in Patent Office: 1,084 Days
• Field of Search: 356/72, 356/73, 356/39, 435/7.24, 436/10, 436/63
• US Class Current: 356/39
• CPC Class Codes: A61B 5/150022 for capillary blood or interstitial fluid
• CPC Class Codes: A61B 5/15003 for venous or arterial blood
• CPC Class Codes: A61B 5/150221 Valves
• CPC Class Codes: A61B 5/150343 Collection vessels for collecting blood samples from the skin surface, e.g. test tubes, cuvettes sampling from within body cavities A61B5/150015, A61B5/150045
• CPC Class Codes: A61B 5/150351 Caps, stoppers or lids for sealing or closing a blood collection vessel or container, e.g. a test-tube or syringe barrel caps for bottles or containers in general B65D41/00
• CPC Class Codes: A61B 5/150809 by audible feedback
• CPC Class Codes: A61B 5/150824 by visual feedback
• CPC Class Codes: A61B 5/157 Devices characterised by integrated means for measuring characteristics of blood
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: G01N 15/1404 Handling flow, e.g. hydrodynamic focusing
• CPC Class Codes: G01N 2015/1486 Counting the particles
• CPC Class Codes: Y10T 436/101666 Particle count or volume standard or control [e.g., platelet count standards, etc.]

Abstract

A portable or wearable cytometer that can be used at remote locations, such as in the field or at home. The flow cytometer of the present invention may help improve the healthcare of many weak, sick or elderly people by providing early detection of infection. By detecting the infection early, the infection may be more readily treatable. In military applications, the portable cytometer of the present invention may help save lives by providing early detection of infection due to biological agents.

Claims

1. A portable cytometer for identifying and/or counting selected particles in a sample fluid, comprising:

a fluid receiver for receiving the sample fluid;

at least one reservoir for storing one or more supporting fluids;

a non-precision fluid driver coupled to the fluid receiver and the at least one reservoir for applying separate pressures to the sample fluid and the one or more supporting fluids to provide a fluid velocity to each of the sample fluid and the one or more supporting fluids;

valve means coupled to the fluid driver for regulating the separate pressures that are applied to the sample fluid and the one or more supporting fluids;

at least one flow sensor for measuring the fluid velocity of the sample fluid and the one or more supporting fluids provided by the fluid driver;

control means coupled to the valve means and the at least one flow sensor for closed-loop control of the valve means so that the fluid velocities of the sample fluid and each of the one or more supporting fluids are at desired levels;

a fluidic circuit for receiving the sample fluid and the one or more supporting fluids from the fluid driver, and for forming a flow stream;

light providing means for providing a light through the flow stream;

light receiving means for receiving the light from the flow stream, 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 identifying and/or counting selected particles therein.

2. A portable cytometer according to claim 1, wherein the non-precision fluid driver includes an electrostatic meso-pump.

3. A portable cytometer according to claim 1, wherein the processing means identifies the velocity of selected particles in the flow stream.

4. A portable cytometer according to claim 1, wherein the processing means identifies the size of selected particles in the flow stream.

5. A portable cytometer according to claim 1, wherein the processing means identifies the alignment of selected particles in the flow stream.

6. A portable cytometer according to claim 5, wherein the processing means provides one or more signals that are used for adjusting the valve means in order to change the relative pressures that are applied to the sample fluid and the one or more supporting fluids to change the alignment of the selected particles in the flow stream.

7. A portable cytometer according to claim 1, wherein said light receiving means comprises an array of detectors situated approximately across the flow stream.

8. A portable cytometer according to claim 7, wherein the non-precision fluid driver is manually powered.

9. A portable cytometer according to claim 8, wherein the non-precision fluid driver includes at least one plunger received within a pressure chamber, wherein the plunger is manually depressed into the pressure chamber.

10. A wearable cytometer for analyzing selected particles in a sample fluid, comprising:

a removable cartridge for receiving the sample fluid and for preparing the sample fluid for analysis including performing hydrodynamic focusing to form a flow stream having a core stream of particles, the cartridge having one or more pressure receiving ports for receiving one or more controlled pressures;

a housing adapted to receive the removable cartridge, said housing having;

pressure applying means for applying a controlled pressure to the one or more pressure receiving ports of the removable cartridge;

one or more light providing means positioned adjacent the flow stream of the removable cartridge for providing light through the flow stream;

two or more light receiving means provided adjacent the flow stream of the removable cartridge for receiving the light from the flow stream, 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 analyzing the at least one signal to determined predetermined information relative to the selected particles in the flow stream.

11. A wearable cytometer according to claim 10, wherein the housing includes:

a base;

a cover; and

a hinge for securing the base to the cover.

12. A wearable cytometer according to claim 10, wherein the removable cartridge and the housing are self aligned using one or more registration pins.

13. A wearable cytometer according to claim 10, wherein the removable cartridge includes:

a fluid receiver for receiving the sample fluid; and

at least one reservoir for storing one or more supporting fluids.

14. A wearable cytometer according to claim 13, wherein the sample fluid is a blood sample, and the one or more supporting fluids include lyse and sheath.

15. A wearable cytometer according to claim 14, wherein the removable cartridge dilutes the blood sample, performs red cell lysing, and performs hydrodynamic focusing for flow and core stream formation.

16. A wearable cytometer according to claim 10, wherein the removable cartridge includes a waste reservoir downstream of the flow stream for receiving and storing the fluid of the flow stream.

17. A wearable cytometer according to claim 10, wherein the pressure applying means includes:

a pressure source for generating an input pressure;

a first pressure chamber for receiving the input pressure;

a second pressure chamber;

first valves means between the first pressure chamber and the second pressure chamber; and

second valve means for relieving the pressure in the second pressure chamber.

18. A wearable cytometer according to claim 17, wherein the pressure source includes a movable plunger that moves into the first pressure chamber.

19. A wearable cytometer according to claim 18, wherein the movable plunger is adapted to be manually depressed.

20. A wearable cytometer according to claim 17, wherein the first valve means comprises a plurality of microvalves.

21. A wearable cytometer according to claim 20, wherein the plurality of microvalves are electrostatically actuated.

22. A wearable cytometer according to claim 21, wherein selected microvalves or groups of microvalves are individually addressable.

23. A wearable cytometer according to claim 17, wherein the second valve means comprises a plurality of microvalves.

24. A wearable cytometer according to claim 17, wherein the replaceable cartridge includes:

a downstream fluid stream in fluid communication with the second pressure chamber;

a flow sensor for measuring the fluid flow in the downstream fluid stream; and

feedback means coupled to the flow sensor and to the first valve means and the second valve means for opening the first valve means when the fluid flow in the downstream fluid stream drops below a first predetermined value and for opening the second valve means when the fluid flow in the downstream fluid stream increases above a second predetermined value.

25. A wearable cytometer according to claim 24, wherein the flow sensor is a thermal anemometer type flow sensor.

26. A wearable cytometer according to claim 25, wherein the thermal anemometer type flow sensor is a microbridge flow sensor.

27. A wearable cytometer according to claim 10, wherein the flow stream has a length along a central axis of flow and a width perpendicular to the central axis of flow.

28. A wearable cytometer according to claim 27, wherein the light providing means includes two or more spaced light sources positioned laterally at different distances from the central axis of flow of the flow stream.

29. A wearable cytometer according to claim 28, wherein the two or more spaced light sources are positioned along a light source axis that is rotated relative to the central axis of flow of the flow stream.

30. A wearable cytometer according to claim 29, wherein the two or more spaced light sources provide substantially constant light intensity across the flow stream.

31. A wearable cytometer according to claim 30, wherein the two or more spaced light sources are part of an array of VCSEL devices.

32. A wearable cytometer according to claim 31, wherein the VCSEL devices operate in the red spectrum.

33. A wearable cytometer according to claim 31, wherein each of the two or more spaced light sources have a corresponding lens.

34. A wearable cytometer according to claim 33, wherein each lens is an integrated micro lens.

35. A wearable cytometer according to claim 33, wherein selected lenses focus the light at a central plane that includes the central axis of the flow stream.

36. A wearable cytometer according to claim 35, wherein the light that is focused at the central plane is used for detecting the small angle scattering (SALS) produced by the selected particles in the flow stream.

37. A wearable cytometer according to claim 35, wherein the light that is focused at the central plane is used for determining the speed of selected particles in the flow stream.

38. A wearable cytometer according to claim 37, wherein the light that is focused at the central plane is used for determining the size of selected particles in the flow stream.

39. A wearable cytometer according to claim 33, wherein selected lenses provide substantially collimated light through the flow stream.

40. A wearable cytometer according to claim 39, wherein the substantially collimated light is used for detecting the forward angle scattering (FALS) produced by the selected particles in the flow stream.

41. A wearable cytometer according to claim 10, wherein the selected particles include white blood cells.

42. A wearable cytometer according to claim 41, wherein the white blood cells include neutrophils and/or lymphocytes white blood cells.

43. A wearable cytometer according to claim 10, wherein the light providing means includes a first set of light sources positioned along a light source axis that is rotated relative to the central axis of flow 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.

44. A wearable cytometer according to claim 43, 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.

45. A wearable cytometer according to claim 44, wherein the first set of light sources and the first set of light detectors are used to detect the alignment of the flow of selected particles relative to the width of the flow stream.

46. A wearable cytometer according to claim 45, wherein the processing means is in communication with the pressure applying means, and adjusts the controlled pressure applied by the pressure applying means to the one or more pressure receiving ports of the removable cartridge so that the alignment of the selected particles in the core stream is substantially in the middle of the flow stream.

47. A wearable cytometer according to claim 43, wherein the first set of light sources and the first set of light detectors are used to detect the velocity of selected particles in the flow stream.

48. A wearable cytometer according to claim 43, wherein the first set of light sources and the first set of light detectors are used to detect the size of selected particles in the flow stream.

49. A wearable cytometer according to claim 43, wherein the light providing means includes a second set of light sources positioned along a light source axis that is rotated relative to the central axis of flow of the flow stream, 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.

50. A wearable cytometer according to claim 49, 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.

51. An wearable cytometer according to claim 50, 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.

52. A wearable cytometer according to claim 51, wherein the second set of light detectors are used to detect the small angle scattering (SALS) produced by the selected particles in the flow stream.

53. A wearable cytometer according to claim 49, wherein the light providing means includes a third set of light sources positioned along a light source axis that is rotated relative to the central axis of flow 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.

54. A wearable cytometer according to claim 53, 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.

55. A wearable cytometer according to claim 54, wherein each of the third set of light detectors are annular in shape.

56. A wearable cytometer according to claim 55, 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.

57. A method for analyzing selected particles in a fluid stream, the method comprising:

manually generating a non-precise pressure;

generating a controlled pressure from the manually generated non-precise pressure;

using the controlled pressure to provide flow to a fluid stream;

adjusting the controlled pressure according to a velocity of the flow of the fluid stream; and

analyzing the selected particles in the fluid stream.

58. A method according to claim 57, wherein the analyzing step includes determining the small angle scattering (SALS) produced by the selected particles in the flow stream.

59. A method according to claim 57, wherein the analyzing step includes determining the forward angle scattering (FALS) produced by the selected particles in the flow stream.

60. A method according to claim 57, wherein the manually generating step includes manually generating two or more non-precise pressures.

61. A method according to claim 60, wherein the generating step includes generating a controlled pressure from each of the two or more manually generated non-precise pressures.

62. A method according to claim 61, wherein each of the controlled pressures provide a force to a different fluid reservoir, wherein each fluid reservoir includes a different fluid.

63. A method according to claim 62, wherein the force that is applied to each fluid reservoir provides a different velocity to each fluid.

64. A method according to claim 63, further comprising performing hydrodynamic focusing using each fluid to form the fluid stream.

65. A method according to claim 64, wherein the analyzing step includes determining the alignment of the selected particles in the fluid stream.

66. A method according to claim 65, further comprising adjusting each of the controlled pressures to provide a desired alignment of the selected particles in the fluid stream.

67. A method according to claim 57, further comprising:

shining a light through the flow stream; and

detecting the light through the flow stream with a plurality of detectors.

68. A method according to claim 67, wherein the analyzing step includes determining the alignment of the selected particles in the fluid stream.