Low acceleration method of flow cytometry
DC CAFC
First Claim
Patent Images
1. A method of flow cytometry sample processing, comprising the steps of:
- a. establishing a sheath fluid;
b. injecting a sample into said sheath fluid at an injection point;
c. subjecting said sample to a first axial motion surface in a nozzle;
d. positioning to a second axial motion surface in said nozzle;
e. subjecting said sample to said second axial motion surface in said nozzle wherein said first and said second axial motion surfaces transition with a maximal acceleration differentiation;
f. coordinating said maximal acceleration differentiation so as to not exceed the practical capabilities of said sample over its length;
g. affirmatively limiting said maximal acceleration differentiation so as to not exceed the practical capabilities of said sample over its length;
h. exiting said sample from said nozzle;
i. analyzing said sample.
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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.
117 Citations
19 Claims
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1. A method of flow cytometry sample processing, comprising the steps of:
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a. establishing a sheath fluid;
b. injecting a sample into said sheath fluid at an injection point;
c. subjecting said sample to a first axial motion surface in a nozzle;
d. positioning to a second axial motion surface in said nozzle;
e. subjecting said sample to said second axial motion surface in said nozzle wherein said first and said second axial motion surfaces transition with a maximal acceleration differentiation;
f. coordinating said maximal acceleration differentiation so as to not exceed the practical capabilities of said sample over its length;
g. affirmatively limiting said maximal acceleration differentiation so as to not exceed the practical capabilities of said sample over its length;
h. exiting said sample from said nozzle;
i. analyzing said sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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−
3 m/sec per micron at a distance of more than 300 um away from the exit orifice,not more than about 50×
10−
6 m/sec per micron at a distance of more than 300 um away from the exit orifice,not more than about 25×
10−
3 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−
6 m/sec per micron2,not more than about 10,000×
10−
6 m/sec per micron2,not more than about 2,000×
1031 6 m/sec per micron2,not more than about 1,100×
10−
6 m/sec per micron2,not more than about 100,000×
10−
6 m/sec per micron2 away from the vicinity of the exit orifice,not more than about 50,000×
10−
6 m/sec per micron2 away from the vicinity of the exit orifice,not more than about 10,000×
10−
6m/sec per micron2 away from the vicinity of the exit orifice,not more than about 5,000×
10−
6 m/sec per micron2 away from the vicinity of the exit orifice,not more than about 1,000×
10−
6 m/sec per micron2 away from the vicinity of the exit orifice,not more than about 300×
10−
6 m/sec per micron2 away from the vicinity of the exit orifice,not more than about 200×
10−
0 m/sec per micron2 at a distance of more than 300 um away from the exit orifice,not more than about 100×
10−
6 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.
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4. A method of flow cytometry sample processing as described in claim 1 wherein said step of transitioning to a second axial motion surface in said nozzle comprises the step of subjecting said sample to a unitary surface.
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5. A method of flow cytometry sample processing as described in claim 1 wherein said step of transitioning to a second axial motion surface in said nozzle comprises the step of subjecting said sample to a unitary exit orifice.
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6. A method of flow cytometry sample processing as described in claim 1 and further comprising the steps of:
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a forming drops around said sample after it has exited said nozzle; and
b. sorting said drops at a rate selected from the group comprising at least 500 sorts per second, at least 1000 sorts per second, and at least 1500 sorts per second.
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7. A method of flow cytometry sample processing as described in claim 1 and further comprising the step of pressurizing said nozzle at a pressure of at least 50 psi.
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8. A method of flow cytometry sample processing as described in claim 6 wherein said step of injecting a sample into said sheath fluid at an injection point comprises the step of injecting sperm cells into said sheath fluid.
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9. A method of flow cytometry sample processing as described in claim 7 wherein said step of injecting a sample into said sheath fluid at an injection point comprises the step of injecting sperm cells into said sheath fluid.
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10. A method of flow cytometry sample processing as described in claim 1, 4, 5, 6, or 7 wherein said step of injecting a sample into said sheath fluid at an injection point comprises the step of injecting sperm cells selected from the group comprising bovine sperm cells and equine sperm cells into said sheath fluid.
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11. A method of creating a sexed sperm specimen comprising the step of producing a sexed sperm specimen as described in any of claim 1, 4, 5, 6, or 7.
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12. A method of creating a mammal comprising the step of producing a sexed sperm specimen as described in any of claim 1, 4, 5, 6, 7, 8, or 9.
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13. A method of flow cytometry sample processing as described in claim 1, 5, 6, 7, or 8 and further comprising the steps of:
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a. establishing a single torsional surface in said nozzle;
b. generating single torsional hydrodynamic forces from said single torsional surface; and
c. orienting said sample with said single torsional hydrodynamic forces.
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14. A method of flow cytometry sample processing as described in claim 13 wherein said single torsional hydrodynamic forces and said maximal acceleration differentiation combine and are affirmatively chosen so as to not exceed the practical capabilities of said sample over its length.
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15. A method of flow cytometry sample processing as described in claim 14 wherein said step of injecting a sample into said sheath fluid at an injection point comprises the step of injecting sperm cells into said sheath fluid.
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16. A method of creating a sexed sperm specimen comprising the step of producing a sexed sperm specimen as described in claim 15 and wherein said step of injecting a sample into said sheath fluid at an injection point comprises the step of injecting sperm cells into said sheath fluid.
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17. A method of creating a sexed sperm specimen comprising the step of producing a sexed sperm specimen as described in claim 16 wherein said step of injecting sperm cells into said sheath fluid comprises the step of injecting sperm cells selected from the group comprising bovine sperm cells and equine sperm cells into said sheath fluid.
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18. A method of creating a mammal comprising the step of producing a sexed sperm specimen as described in claim 15 and wherein said step of injecting a sample into said sheath fluid at an injection point comprises the step of injecting sperm cells into said sheath fluid.
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19. A method of creating a mammal comprising the step of producing a sexed sperm specimen as described in claim 18 wherein said step of injecting sperm cells into said sheath fluid comprises the step of injecting sperm cells selected from the group comprising bovine sperm cells and equine sperm cells into said sheath fluid.
Specification
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Litigation Data
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Current AssigneeXY, LLC (Inguran LLC)
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Original AssigneeXY Incorporated (Inguran LLC)
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InventorsHerickhoff, Lisa A., Buchanan, Kris S.
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Primary Examiner(s)RAEVIS, ROBERT R
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Application NumberUS10/098,036Publication NumberTime in Patent Office517 DaysField of Search738/658, 738/655, 324/71.4, 356/440-442, 422/73, 436/10, 436/63, 435/287.1US Class Current73/865.5CPC Class CodesG01N 15/01 specially adapted for biolo...G01N 15/1404 Handling flow, e.g. hydrody...G01N 15/1409 Handling samples, e.g. inje...G01N 15/149 specially adapted for sorti...G01N 2015/1006 for cytologyG01N 2015/1413 Hydrodynamic focussingG01N 2015/1415 Control of particle positionY10T 436/101666 Particle count or volume st...
