System for analyzing particles using multiple flow cytometry units

• US 7,758,811 B2
• Filed: 03/29/2004
• Issued: 07/20/2010
• Est. Priority Date: 03/28/2003
• Status: Active Grant

• Current Assignee: Inguran LLC
• Original Assignee: Inguran LLC
• Inventors: Durack, Gary, Westfall, Lon A., Vandre, Gary P., Nayak, Niraj V., Wallace, Jeffrey D., Hatcher, Jeremy T.
• Primary Examiner(s): WALLENHORST, MAUREEN
• Application Number:
  US10/812,351
• Publication Number:
  US 20050112541A1
• Time in Patent Office: 2,304 Days
• Field of Search: 436/63, 436/164, 436/165, 436/43, 436/50, 436/55, 436/172, 422/67, 422/68.1, 422/73, 422/82.05, 422/82.08, 435/2, 435/287.1
• US Class Current: 422/73
• CPC Class Codes: A01N 1/0284 Temperature processes, i.e. using a designated change in temperature over time
• CPC Class Codes: C12N 5/06 Animal cells or tissues; Human cells or tissues preservation of living cells or tissues A01N1/02
• CPC Class Codes: C12N 5/061 Sperm cells, spermatogonia
• CPC Class Codes: C12N 5/0612 sorting of gametes, e.g. according to sex or motility
• CPC Class Codes: C12Q 1/02 involving viable microorganisms
• CPC Class Codes: C12Q 3/00 Condition responsive control processes apparatus therefor C12M1/36; controlling or regulating in general G05
• 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 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/1468 with spatial resolution of the texture or inner structure of the particle
• CPC Class Codes: G01N 15/147 the analysis being performed on a sample stream
• 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 2015/1406 Control of droplet point
• CPC Class Codes: G01N 2015/1415 Control of particle position
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 21/63 optically excited
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 33/48 Biological material, e.g. blood, urine G01N33/02, G01N33/26, G01N33/44, G01N33/46 take precedence; Haemocytometers counting blood corpuscules distributed over a surface by scanning the surface G06M11/02
• CPC Class Codes: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025
• CPC Class Codes: Y10T 436/10 : Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• CPC Class Codes: Y10T 436/115831 : Condition or time responsive
• CPC Class Codes: Y10T 436/12 : Condition responsive control
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/2525 : Stabilizing or preserving
• CPC Class Codes: Y10T 436/25375 : Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/255 : including use of a solid sorbent, semipermeable membrane, or liquid extraction
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

A multichannel system for classifying particles according to one or more characteristics of the particles includes a plurality of flow cytometry units. Each flow cytometry unit is operable to classify particles in a mixture of particles by interrogating a stream of fluid containing particles using a beam of electromagnetic radiation. Each flow cytometry unit has a sensor operable to generate a time-varying output signal indicative of at least one characteristic of the particles in the stream of fluid as the stream of fluid is interrogated by the beam of electromagnetic radiation. The flow cytometry units share an integrated platform and perhaps even a common processor for receiving and processing information from the units. The common processor is programmed to receive the time-varying output signals from the flow cytometry units substantially continuously and to process the output signals.

Claims

1. A multi-channel system for classifying sperm cells according to one or more characteristics of the sperm cells, said system comprising:

a plurality of flow cytometry units each of which is operable to provide sex sorted sperm from a mixture of sperm cells by interrogating a stream of fluid containing said sperm cells using a beam of pulsed electromagnetic radiation,

at least one sensor operable to generate a time-varying output signal indicative of at least one characteristic of the sperm cells;

said units sharing an integrated platform comprising a common source of said pulsed electromagnetic radiation and a common processor for receiving and processing at least one output signal from the at least one sensor as a substantially continuous stream and to process the output signals in real time to send control signals to the flow cytometry units during a sorting process to adjust a sort operation as a function of said output signals received by the common processor.

2. The system of claim 1 wherein said common source of said pulsed electromagnetic radiation comprises a plurality of laser pulses, each pulse having a peak power that is greater than the average power output of a laser and a wavelength in a range of about 350-370 nm.

3. The system of claim 2 wherein said laser emits pulses having a width of about 1-100 picoseconds at a pulse frequency of about 50-150 MHz at a power of about 50-500 milliwatts.

4. The system of claim 3 wherein said sex sorted sperm has an enriched subpopulation having a coefficient of variation selected from a group consisting of:

between about 2.0% and about 1.0%, between about 1.5% and about 1.0%, about 1.4%, and about 1.3%.

5. The system of claim 1 wherein said common source of said pulsed electromagnetic radiation comprises a single pulsed laser beam.

6. The system of claim 5 further comprising a beam splitting system for splitting the single pulsed laser beam into multiple beams and directing the multiple beams into optics systems of respective flow cytometry units.

7. The system of claim 1 wherein said integrated platform further comprises a common housing, said flow cytometry units comprising interchangeable modules removably mounted in the housing.

8. The system of claim 1 wherein each flow cytometry unit comprises an epi-illumination optics system for interrogating a respective fluid stream.

9. The system of claim 1 wherein said processor is operable to output an indication of the fluorescence intensity measured by each unit.

10. The system of claim 1 wherein said processor is operable to output an indication of the rate at which each unit is separating sperm cells.

11. The system of claim 1 wherein said processor is operable to output an indication of sperm cell staining variations.

12. The system of claim 1 wherein said processor is operable to output an indication of a decision boundary used by each unit for discriminating between sperm cells.

13. The system of claim 1 wherein said flow cytometry units are adapted to operate in parallel.

14. The system of claim 1 wherein said integrated platform further comprises a common input for controlling operation of the units.

15. The system of claim 14 wherein said plurality of flow cytometry units comprises a jet-in-air droplet sorting flow cytometry unit.

16. The system of claim 1 wherein the common processor receives and processes said information to permit evaluation of the operation of one unit relative to another unit.

17. The system of claim 1 wherein the processor is operable to process the information in real time.

18. The system of claim 1 wherein said integrated platform further comprises at least one of:

(1) a common supply of sperm cells;

(2) a common housing;

(3) a common input for controlling operation of the units; and

(4) a common fluid delivery system for delivering fluid containing said sperm cells to said flow cytometry units.