Miniaturized cytometer for detecting multiple species in a sample

• US 7,630,063 B2
• Filed: 09/09/2004
• Issued: 12/08/2009
• Est. Priority Date: 08/02/2000
• Status: Active Grant

• Current Assignee: Honeywell International Inc.
• Original Assignee: Honeywell International Inc.
• Inventors: Padmanabhan, Aravind, Fritz, Bernard S.
• Primary Examiner(s): Nguyen; Sang
• Application Number:
  US10/938,265
• Publication Number:
  US 20050105077A1
• Time in Patent Office: 1,916 Days
• Field of Search: 356 27- 28, 356317-334, 356/39, 356432-444, 356/246, 356336-337, 436/52, 436/63, 436/173, 436/523, 436/531, 422/82.08
• US Class Current: 356/39
• CPC Class Codes: A61B 5/150022 for capillary blood or interstitial fluid
• CPC Class Codes: A61B 5/150213 Venting means
• CPC Class Codes: A61B 5/150221 Valves
• CPC Class Codes: A61B 5/150274 Manufacture or production processes or steps for blood sampling devices
• 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 5/157 Devices characterised by integrated means for measuring characteristics of blood
• CPC Class Codes: A61B 5/417 the bone marrow
• CPC Class Codes: A61B 5/7264 Classification of physiological signals or data, e.g. using neural networks, statistical classifiers, expert systems or fuzzy systems neural networks per se G06N3/00; expert systems per se G06N5/00
• 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: G01B 11/272 using photoelectric detection means
• CPC Class Codes: G01N 15/1404 Handling flow, e.g. hydrodynamic focusing
• CPC Class Codes: G01N 15/1434 Optical arrangements
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/1484 microstructural devices
• 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 2015/1486 : Counting the particles
• CPC Class Codes: G01N 33/5094 : for blood cell populations red blood cells G01N33/80
• CPC Class Codes: G01N 33/569 : for microorganisms, e.g. protozoa, bacteria, viruses
• CPC Class Codes: G01N 35/085 : Flow Injection Analysis
• CPC Class Codes: Y10T 436/24 : Nuclear magnetic resonance, electron spin resonance or other spin effects or mass spectrometry

Abstract

A system for scattered light and simultaneous multi-color (e.g., greater than sixteen colors) fluorescence light detecting, and for analyzing, classifying and identifying biological particles and items of interest. A sample to be tested may be entered in a disposable microfluidic cartridge which in turn is insertable in a portable, hand-holdable, or wearable miniaturized cytometer instrument. The present system may be incorporated in the cytometer instrument. It may have significant application relative to biological warfare, environmental substances, the medical field and other fields.

Claims

1. A system comprising:

a fluidic channel disposed in a removable miniature cartridge;

a light source arrangement proximate to the fluidic channel;

a wavelength discriminator proximate to the fluidic channel; and

a light detector arrangement including a plurality of detectors proximate to the wavelength discriminator, the plurality of detectors situated in a miniaturized package; and

wherein;

the cartridge is credit-card sized; and

the light source arrangement comprises;

a first light source for impinging the fluidic channel with a blue light; and

a second light source for impinging the fluidic channel with a red light.

2. The system of claim 1, wherein the system is a flow cytometer.

3. The system of claim 1, wherein the system is a hematology analyzer.

4. The system of claim 1, wherein the light detector arrangement provide a plurality of electrical signals wherein each signal represents a wavelength range of detected light from the fluidic channel.

5. The system of claim 4, wherein the light detector arrangement comprises a fluorescent light detector arrangement.

6. The system of claim 4, wherein the light detector arrangement comprises a light scatter detector arrangement proximate to the fluidic channel.

7. The system of claim 6, wherein the light scatter detector arrangement comprises a FALS detector.

8. The system of claim 7, wherein the FALS detector is an annular detector.

9. The system of claim 7, wherein the light scatter detector arrangement further comprises a SALS detector.

10. The system of claim 4, wherein the light source arrangement comprises at least one light source having a wavelength greater than 700 nanometers.

11. The system of claim 4, wherein the light source arrangement comprises at least one light source having a wavelength approximately between 450 nanometers and 700 nanometers.

12. The system of claim 4, wherein the light source arrangement comprises at least one light source having a wavelength less than 450 nanometers.

13. The system of claim 4, wherein the wavelength discriminator comprises a prism.

14. The system of claim 4, wherein the wavelength discriminator comprises one or more filters.

15. The system of claim 14, wherein the filters are band-pass filters.

16. The system of claim 4, wherein the wavelength discriminator comprises a dispersive element.

17. The system of claim 4, wherein the wavelength discriminator comprises a wavelength demultiplexer.

18. The system of claim 4, wherein the wavelength discriminator comprises an optical grating.

19. The system of claim 4, wherein the wavelength discriminator comprises an arrayed waveguide grating.

20. The system of claim 4, wherein the wavelength discriminator comprises one or more dichroic mirrors.

21. The system of claim 1, wherein the detectors of the light detector arrangement may approximately simultaneously detect various wavelengths of light.

22. The system of claim 21, wherein the light detector arrangement may detect a plurality of colors greater than seven.

23. The system of claim 21, wherein the light detector arrangement may detect a plurality of colors greater than sixteen.

24. The system of claim 21, wherein the light detector arrangement may detect a plurality of colors greater than thirty-two.

25. The system of claim 1, wherein the detectors are avalanche photo detectors.

26. The system of claim 1, wherein the detectors are at least one charge coupled discharge (CCD) array.

27. The system of claim 1, wherein the detectors are silicon photo detectors.

28. The system of claim 1, wherein the detectors are photo multiplier tubes.

29. The system of claim 1, wherein the fluidic channel is for a core stream of blood cells.

30. The system of claim 1, wherein the fluidic channel is for a core stream of biological particles.

31. The system of claim 1, wherein the cartridge comprises a low autofluorescence non-glass material.

32. The system of claim 31, wherein the non-glass material is a cyclic olefin copolymer based plastic.

33. The system of claim 1, wherein the system comprises characteristics of good sensitivity, quantification of agent concentration, high specificity of detection of bacteria, viruses, spores, toxins and/or other like particles, automated sample preparation, very small handheld unit size and weight, and low power consumption.

34. The system of claim 1, wherein the light source arrangement and the wavelength discriminator are situated in the miniaturized package.

35. A method comprising:

providing a flow path in a miniature disposable cartridge;

projecting two or more different wavelengths of light on the flow path of the miniature disposable cartridge;

detecting light according to wavelength from the flow path, where said detecting involves parallel channels of a plurality of wavelength discriminators; and

obtaining information on particles in the flow path from the detected light; and

wherein the cartridge is credit-card sized.

36. The method of claim 35, further comprising identifying species of particles in the flow path approximately simultaneously according to wavelength of the light detected from the flow path.

37. The method of claim 36, further comprising classifying the particles according to wavelength of the detected light.

38. The method of claim 36, wherein the detecting light according to wavelength from the flow path is performed with a plurality of detectors in a miniaturized package.

39. The method of claim 35, further comprising detecting scattered light from the flow path.

40. The method of claim 39, further comprising counting particles in the flow path.

41. The method of claim 35, wherein the particles in the flow path are blood cells.

42. The method of claim 41, wherein the particles in the flow path are biological particles.

43. The method of claim 35, wherein the flow path is within a fluidic channel of a flow cytometer.

44. The method of claim 35, wherein the flow path is within a fluidic channel of a hematology analyzer.

45. The method of claim 35, wherein the cartridge comprises a low autofluorescence non-glass material.

46. The method of claim 45, wherein the non-glass material is a cyclic olefin copolymer based plastic.

47. A device for cytometry comprising:

a disposable miniature cartridge including a fluidic channel that is configured to hydro-dynamically focus particles in the flow channel into a single file arrangement;

at least two light sources positioned to illuminate the particles in the fluidic channel, wherein the disposable miniature cartridge is credit-card sized; and

a microspectrometer configured for detecting light from the channel according to wavelength; and

wherein;

a first light source emits red light; and

a second light source emits blue light.

48. The device of claim 47,

further comprising a wavelength discriminator, wherein the microspectrometer includes a plurality of detectors in a linear detector array;

the plurality of detectors is situated in a miniaturized package; and

the microspectrometer provides approximately simultaneous detection for all of the detected wavelengths.

49. The device of claim 48, wherein the microspectrometer detects fluorescent light from markers associated with various types of the particles in the channel.

50. The device of claim 49, wherein the markers associated with the various types of particles emit light different wavelengths so that each type of particle may be identified according to the wavelength emitted by the marker.

51. The device of claim 50, wherein the markers comprise phosphors.

52. The device of claim 50, wherein the markers are fluorescent dyes.

53. The device of claim 48, wherein the device is a flow cytometer.

54. The device of claim 48, wherein the device is a hematology analyzer.

55. The device of claim 49, further comprising a detector for detecting scattered light from the channel.

56. The device of claim 47, wherein the fluidic channel is for a core stream of blood cells.

57. The device of claim 47, wherein the fluidic channel is for a core stream of biological particles.

58. The device of claim 47, wherein the fluidic cartridge comprises a low autofluorescence non-glass material.

59. The device of claim 58, wherein the non-glass material is a cyclic olefin copolymer based plastic.