Flow cytometer nozzle and flow cytometer sample handling methods

• US 6,263,745 B1
• Filed: 12/03/1999
• Issued: 07/24/2001
• Est. Priority Date: 12/03/1999
• Status: Expired due to Term

• Current Assignee: XY, LLC (Inguran LLC)
• Original Assignee: XY Incorporated (Inguran LLC)
• Inventors: Herickhoff, Lisa A., Buchanan, Kris S.
• Primary Examiner(s): RAEVIS, ROBERT R
• Application Number:
  US09/454,488
• Time in Patent Office: 599 Days
• Field of Search: 738/655, 324/71.4, 356/440-442
• US Class Current: 73/865.5
• CPC Class Codes: G01N 15/01 specially adapted for biological cells, e.g. blood cells investigating sedimentation of particle suspensions in blood G01N15/05
• CPC Class Codes: G01N 15/1404 Handling flow, e.g. hydrodynamic focusing
• CPC Class Codes: G01N 15/1409 Handling samples, e.g. injecting samples
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2015/1006 for cytology
• CPC Class Codes: G01N 2015/1413 Hydrodynamic focussing
• CPC Class Codes: G01N 2015/1415 Control of particle position
• CPC Class Codes: Y10T 436/101666 Particle count or volume standard or control [e.g., platelet count standards, etc.]

Abstract

An improved nozzle system for a flow cytometer and accompanying methods have been invented for a high efficiency orientation and sorting process of a flat sample and dedicates items such as equine or bovine sperm cells. This improved nozzle system comprises a nozzle with a novel interior surface geometry that can both gently accelerate the cells and can include an elliptical-like, single torsional interior surface element within the nozzle, i.e., a single torsional orientation nozzle. The elliptical-like, single torsional interior surface element may have a laminar flow surface and may produce the simplest flow path for applying minimal forces which act in either an accelerative nature or orienting hydrodynamic forces, namely, the single torsional orientation forces, to orient a flat sample such as animal sperm cells into a proper direction for an analyzing and efficiently sorting process in clinical use, for research and for the animal insemination industry.

Claims

1. A flow cytometer system, comprising:

a. a sample injection tube having an injection point through which a sample may be introduced;

b. a sheath fluid container having a bottom end and wherein said sample injection tube is located within said sheath fluid container;

c. a sheath fluid port connected to said sheath fluid container;

d. a single torsional orientation nozzle located at least in part below said injection point; and

e. an analytical system which senses below said single torsional orientation nozzle.

2. A flow cytometer system as described in claim 1 wherein said single torsional orientation nozzle comprises a single torsional interior surface element.

3. A flow cytometer system as described in claim 2 wherein said single torsional interior surface element comprises a tapered single torsional interior surface element and wherein said tapered single torsional interior surface element is of an ellipse-shape or an oval-shape.

4. A flow cytometer system as described in claim 1 and further comprising:

a. a first axial motion surface in said nozzle;

b. a second axial motion surface in said nozzle; and

c. a limited maximal acceleration differentiation transition area between said first axial motion surface in said nozzle and said second axial motion surface in said nozzle wherein said limited maximal acceleration differentiation transition area is coordinated with said sample so as to be affirmatively limited to maintain a functional sample over its length.

5. A flow cytometer system as described in claim 4 wherein said first axial motion surface comprises a first axial acceleration surface and wherein said second axial motion surface comprises a second axial acceleration surface.

6. A flow cytometer system as described in claim 5 wherein said nozzle has acceleration values caused by its internal surface and wherein said acceleration values are selected from a group comprising:

not more than about 0.16 m/sec per micron,

not more than about 0.05 m/sec per micron away from the vicinity of the exit orifice,

not more than about 0.10 m/sec per micron away from the vicinity of the exit orifice,

not more than about 0.13 m/sec per micron away from the vicinity of the exit orifice,

not more than about 0.16 m/sec per micron in the vicinity of the exit orifice,

not more than about 0.20 m/sec per micron in the vicinity of the exit orifice,

not more than about 0.23 m/sec per micron in the vicinity of the exit orifice,

not more than about 100×

10<sup>−

3</sup> m/sec per micron at a distance of more than 300 um away from the exit orifice,

not more than about 50×

10<sup>−

3</sup> m/sec per micron at a distance of more than 300 um away from the exit orifice,

not more than about 25×

10<sup>−

3</sup> m/sec per micron at a distance of more than 300 um away from the exit orifice,

such acceleration values with respect to axial location as do not discontinuously change along a central axis,

not more than about 100,000×

10<sup>−

6</sup> m/sec per micron²,

not more than about 100,000×

10<sup>−

6</sup> m/sec per micron²,

not more than about 2,000×

10<sup>−

6</sup> m/sec per micron²,

not more than about 1,100×

10<sup>−

6</sup> m/sec per micron²,

not more than about 100,000×

10<sup>−

6</sup> m/sec per micron² away from the vicinity of the exit orifice,

not more than about 50,000×

10<sup>−

6</sup> m/sec per micron² away from the vicinity of the exit orifice,

not more than about 10,000×

10<sup>−

6</sup> m/sec per micron² away from the vicinity of the exit orifice,

not more than about 5,000×

10<sup>−

6</sup> m/sec per micron² away from the vicinity of the exit orifice,

not more than about 1,000×

10<sup>−

6</sup> m/sec per micron² away from the vicinity of the exit orifice,

not more than about 300×

10<sup>−

6</sup> m/sec per micron² away from the vicinity of the exit orifice,

not more than about 200×

10<sup>−

6</sup> m/sec per micron² at a distance of more than 300 um away from the exit orifice,

not more than about 100×

10<sup>−

6</sup> m/sec per micron² at a distance of more than 300 um away from the exit orifice,

such rate of change of acceleration values with respect to axial location as do not discontinuously change along a central axis, and

such rate of change of acceleration values with respect to axial location as do not change sign along a central axis away from the vicinity of the exit orifice.

7. A flow cytometer system as described in claim 4 wherein said limited maximal acceleration differentiation transition area comprises a unitary surface.

8. A flow cytometer system as described in claim 4 wherein said limited maximal acceleration differentiation transition area comprises a unitary exit orifice.

9. A flow cytometer system as described in claim 4 wherein said analytical system which senses below said nozzle operates at a rate selected from a group comprising at least 500 sorts per second, at least 1000 sorts per second, and at least 1500 sorts per second.

10. A flow cytometer system as described in claim 4 and further comprising a pressurization system to which said flow cytometer system is responsive and which operates at least about 50 psi.

11. A flow cytometer system as described in claim 9 and further comprising a sperm collection system.

12. A flow cytometer system as described in claim 10 and further comprising a sperm collection system.

13. A flow cytometer system as described in claim 4, 7, 8, 9, or 10 wherein said sample comprises sperm cells selected from a group comprising bovine sperm cells and equine sperm cells.

14. A sexed sperm specimen produced with a flow cytometer system as described in claim 11 or 12.

15. A flow cytometer system as described in claim 14 wherein said sample comprises sperm cells selected from a group comprising bovine sperm cells and equine sperm cells.

16. A mammal produced through use of a sexed sperm specimen produced with a flow cytometer system as described in claim 11 or 12.

17. A flow cytometer system as described in claim 16 wherein said sample comprises sperm cells selected from a group comprising bovine sperm cells and equine sperm cells.

18. A flow cytometer system as described in claim 3 wherein said tapered, elliptical-like, single torsional interior surface element comprises:

a. an ellipse-like demarcation location located at about said injection point; and

b. an ellipticity-decreasing zone extending from below said ellipse-like demarcation location.

19. A flow cytometer system as described in claim 3 wherein said tapered, elliptical-like, single torsional interior surface element comprises:

a. an ellipticity-increasing zone;

b. an ellipse-like demarcation location downstream from said ellipticity-increasing zone; and

c. an ellipticity-decreasing zone extending from said ellipse-like demarcation location.

20. A flow cytometer system as described in claim 18 and further comprising:

a. a conical zone located below said ellipticity-decreasing zone;

b. a cylindrical zone located below said conical zone;

wherein both said conical zone and said cylindrical zone comprises a laminar flow surface;

c. a circular exit orifice located below said cylindrical zone;

d. an oscillator to which said circular exit orifice is responsive; and

e. a flow cytometry sorting system below said single torsional orientation nozzle.

21. A flow cytometer system as described in claim 19 and further comprising:

a. a conical zone located below said ellipticity-decreasing zone;

b. a cylindrical zone located below said conical zone;

wherein both said conical zone and said cylindrical zone comprises a laminar flow surface; and

c. a circular exit orifice located below said cylindrical zone;

d. an oscillator to which said circular exit orifice is responsive; and

e. a flow cytometry sorting system below said single torsional orientation nozzle.

22. A flow cytometer system as described in claims 18 wherein said tapered, elliptical-like, single torsional interior surface element, said conical zone, and said cylindrical zone are unitary.

23. A flow cytometer system as described in claims 20 wherein said tapered, elliptical-like, a single torsional interior surface element, said conical zone, said cylindrical zone, and said circular exit orifice are unitary.

24. A flow cytometer system as described in claim 22 wherein said ellipse-like demarcation location has a major axis and a minor axis having a ratio, and wherein said ratio of said major axis to minor axis comprises a ratio which maintains a functional sample.

25. A flow cytometer system as described in claim 24 wherein said major axis of said desired ellipse-like demarcation location is about 2.2 mm and wherein said minor axis of said desired ellipse-like demarcation location is about 1.0 mm.

26. A flow cytometer system as described in claim 19 wherein said ellipse-like demarcation location has a major axis and a minor axis, wherein said major axis of said desired ellipse-like demarcation location is about 2.2 mm and wherein said minor axis of said desired ellipse-like demarcation location is about 1.0 mm.

27. A flow cytometer system as described in claim 22 wherein said single torsional orientation nozzle has a downstream direction, wherein said ellipticity-decreasing zone has cross sections and cross section areas, wherein said cross sections of said ellipticity-decreasing zone undergo transitional changes from ellipse-like shapes to circular shapes downstream, and wherein said cross section areas become progressively smaller downsteam.

28. A flow cytometer system as described in claim 27 wherein each of said cross sections of said ellipticity-decreasing zone has a major axis and a minor axis and wherein said major axis and said minor axis progressively become equal downstream.

29. A flow cytometer system as described in claim 21 wherein said conical zone is about 0.3 mm in height.

30. A flow cytometer system as described in claim 29 herein said cylindrical zone is about 0.15 mm in height.

31. A flow cytometer system as described in claim 2 wherein said single torsional interior surface element comprises a gradually tapered, single torsional interior surface element.

32. A flow cytometer system as described in claim 31 wherein said gradually tapered, single torsional interior surface element comprises an interior surface element that tapers at about 23°

.

33. A flow cytometer system as described in claim 19 wherein said tapered, single torsional interior surface element comprises an interior surface element that tapers at about 23°

.

34. A flow cytometer system as described in claim 23 wherein said single torsional orientation nozzle comprises a single torsional, ceramic orientation nozzle.

35. A flow cytometer system as described in claim 22 wherein said single torsional orientation nozzle has a height and a top with an outer diameter, and wherein said height is about 13 mm, and wherein said outer diameter about 6 mm.

36. A flow cytometer system as described in claim 20 wherein said flow cytometer system comprises a circular exit orifice, and wherein said tapered, elliptical-like, single torsional interior surface element has a mouth and wherein said mouth is about 5.25 mm in diameter and said circular exit orifice is about 0.07 mm in diameter.

37. A flow cytometer system as described in claim 35 wherein said flow cytometer system comprises a circular exit orifice, and wherein said tapered, elliptical-like, single torsional interior surface element has a mouth and wherein said mouth is about 5.25 mm in diameter and said circular exit orifice is about 0.07 mm in diameter.

38. A flow cytometer system as described in claim 36 wherein said conical zone has a top with an inner diameter, and wherein said inner diameter at said top of said conical zone is about 0.19 mm.

39. A flow cytometer system as described in claim 37 wherein said conical zone has a top with an inner diameter, and wherein said inner diameter at said top of said conical zone is about 0.19 mm.

40. A flow cytometer system as described in claim 18 wherein said sample injection tube comprises a sample orientation tube.

41. A flow cytometer system as described in claim 40 wherein said orientation-improving sample injection tube comprises a beveled tip.

42. A flow cytometer system as described in claim 41 wherein said beveled tip has a circular mouth and wherein said circular mouth has a diameter of about 0.01 mm.

43. A flow cytometer system as described in claim 41 wherein said tapered, elliptical-like interior zone has a major axis and a minor axis at said injection point, and wherein said major axis of said beveled tip is aligned with said major axis of said tapered, elliptical-like interior zone at said injection point.

44. A flow cytometer system as described in claim 43 wherein said single torsional orientation nozzle has a bottom, wherein said beveled tip has a circular mouth, wherein said flow cytometer system further comprises a circular exit orifice located at said bottom of said single torsional orientation nozzle, and wherein said injection point is located at a distance from said circular exit orifice of said single torsional orientation nozzle at which a sample exiting from said circular mouth of said beveled tip receives minimal torquing forces to achieve an orientationally aligned status.

45. A flow cytometer system as described in claim 44 wherein said injection point is located at a distance from said circular exit orifice at which said orientationally aligned status of said sample is substantially maintained when said sample exits said circular orifice of said single torsional orientation nozzle.

46. A flow cytometer system as described in claim 45 wherein said injection point is located about 6 mm from said circular exit orifice of said single torsional orientation nozzle.

47. A flow cytometer system as described in claim 9 wherein said flow cytometer system has dimensions established according to claims 26, 29, 30, 36, 38 or 46.

48. A flow cytometer system as described in claim 1 wherein said sample comprises sperm cells in a sperm compatible buffer.

49. A flow cytometer system as described in claim 48 wherein said analytical system comprises a flow cytometry sorting system.

50. A flow cytometer system as described in claim 49 and further comprising a sperm compatible collection system.

51. A flow cytometer system as described in claim 49 wherein said sample comprises sperm cells in a sperm compatible buffer and wherein said sperm cells are selected from a group consisting of equine sperm cells and bovine sperm cells.

52. A flow cytometer system as described in claim 51 wherein said flow cytometer system has dimensions established according to claims 26, 29, 30, 36, 38, or 46.

53. A sexed sperm specimen produced with a flow cytometer system as described in any of claims 1, 20, 23, 24, 26, 29, 30, 32, 39, 41, 44, 45, 51, or 52.

54. A mammal produced through use of a sexed sperm specimen produced with a flow cytometer system as described in any of claims 1, 20, 23, 24, 26, 29, 30, 32, 39, 41, 44, 45, 51, or 52.

55. A flow cytometer system as described in claim 18, 23, 26, 29, 30, 32, 37, 39, 42, 46 and further comprising:

a. a first axial motion surface in said nozzle;

b. a second axial motion surface in said nozzle; and

c. a limited maximal acceleration differentiation transition area between said first axial motion surface in said nozzle and said second axial motion surface in said nozzle wherein said limited maximal acceleration differentiation transition area is coordinated with said sample so as to be affirmatively limited to not exceed the practical capabilities of said sample over its length.

56. A flow cytometer system as described in claim 55 wherein said limited maximal acceleration differentiation transition area comprises a unitary surface.

58. A flow cytometer system as described in claim 55 wherein said analytical system which senses below said nozzle operates at a rate selected from a group comprising at least 500 sorts per second, at least 1000 sorts per second, and at least 1500 sorts per second.

59. A flow cytometer system as described in claim 55 and further comprising a pressurization system which operates at least about 50 psi.

60. A flow cytometer system as described in claim 58 and further comprising a sperm collection system.

61. A flow cytometer system as described in claim 59 and further comprising a sperm collection system.

62. A flow cytometer system as described in claim 55 wherein a said sample comprises sperm cells selected from a group comprising bovine sperm cells and equine sperm cells.

64. A flow cytometer system as described in claim 58 wherein a said sample comprises sperm cells selected from a group comprising bovine sperm cells and equine sperm cells.

65. A flow cytometer system as described in claim 59 wherein a said sample comprises sperm cells selected from a group comprising bovine sperm cells and equine sperm cells.

66. A sexed sperm specimen produced with a flow cytometer system as described in claim 60.

67. A sexed sperm specimen produced with a flow cytometer system as described in claim 64.

68. A mammal produced through use of a sexed sperm specimen produced with a flow cytometer system as described in claim 60.

69. A mammal produced through use of a sexed sperm specimen produced with a flow cytometer system as described in claim 64.

57. A flow cytometer system as described in claim herein said limited maximal acceleration differentiation transition area comprises a unitary exit orifice.

63. A flow cytometer system as described in claim 57 wherein a said sample comprises sperm cells selected from a group comprising bovine sperm cells and equine sperm cells.