Apparatus, methods and processes for sorting particles and for providing sex-sorted animal sperm

• US 9,377,390 B2
• Filed: 04/10/2015
• Issued: 06/28/2016
• Est. Priority Date: 03/28/2003
• Status: Active Grant

• Current Assignee: Inguran LLC
• Original Assignee: Inguran LLC
• Inventors: Durack, Gary, Wallace, Jeffrey D., Vandre, Gary P., Westfall, Lon A., Hatcher, Jeremy T., Nayak, Niraj V.
• Primary Examiner(s): Wallenhorst, Maureen
• Application Number:
  US14/683,936
• Publication Number:
  US 20150211978A1
• Time in Patent Office: 445 Days
• Field of Search: 422/67, 422/68.1, 422/73, 422/82.05, 422/82.08, 435/2, 435/29, 435/34, 435/366, 435/286.1, 435/287.1, 435/288.7, 436/43, 436/50, 436/55, 436/63, 436/164, 436/165, 436/172
• US Class Current: 1/1
• 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 multi-channel system for classifying particles in a mixture of particles according to one or more characteristics including a common source of electromagnetic radiation for producing a beam of electromagnetic radiation and a beam splitter for producing multiple beams of electromagnetic radiation for directing multiple beams of electromagnetic radiation to each interrogation location associated with each flow channel of the multi-channel system.

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 channels, each of which directs sample fluid containing a mixture of sperm cells through an interrogation location when in use;

a common fluid source in communication with each of the plurality of flow channels for supplying sheath fluid to surround the sample fluid in each of the plurality of flow channels;

a common source of electromagnetic radiation for producing a beam of electromagnetic radiation;

a beam splitter for producing multiple beams of electromagnetic radiation from the common source of electromagnetic radiation, wherein at least one of the multiple beams of electromagnetic radiation intersects the interrogation location associated with each flow channel;

at least one detector which detects electromagnetic radiation from sperm cells at each interrogation location;

said detector being operable to output a time-varying analog signal indicative of the intensity of the detected electromagnetic radiation;

an analog to digital converter adapted to receive the time-varying analog signal as input and to sample the analog signal to produce a digitized output; and

a processor operable to receive the digitized output and classify sperm cells in the mixture of sperm cells according to one or more characteristics of the sperm cells.

2. The multi-channel system of claim 1, wherein each flow channel is associated with a flow cytometry unit in a plurality of flow cytometry units.

3. The multi-channel system of claim 2, wherein each flow cytometry unit is housed in a common housing.

4. The multi-channel system of claim 2, wherein said flow cytometry units comprise interchangeable modules removably mounted in the common housing.

5. The multi-channel system of claim 2, wherein said flow cytometry units are configured to operate in parallel.

6. The multi-channel system of claim 2, wherein said plurality of flow cytometry units are operable to sort the sperm cells.

7. The multi-channel system of claim 2, wherein said plurality of flow cytometry units comprises a jet-in air droplet sorting flow cytometry unit.

8. The multi-channel system of claim 2, wherein one or more of the plurality of flow cytometry units comprises a jet-in-air droplet sorting flow cytometry unit.

9. The multi-channel system of claim 2, further comprising a common input for controlling operation of said plurality of flow cytometry units and wherein the processor comprises a common processor for receiving and processing information from said plurality of flow cytometry units.

10. The multi-channel system of claim 9 wherein the common processor is operable to send control signals to the plurality of flow cytometry units in real time during a sorting process to adjust their operation as a function of said information received by the common processor, and wherein the flow cytometry units are responsive to the control signals.

11. The multi-channel system of claim 2, wherein at least one of said plurality of flow cytometry units comprises a photo-damage system.

12. The multi-channel system of claim 1, wherein the processor comprises a common processor which receives digitized output from sperm cells in each of the flow channels, wherein each flow channel is associated with a flow cytometry unit in a plurality of flow cytometry units.

13. The multi-channel system of claim 12, wherein the common processor comprises one or more digital signal processors.

14. The multi-channel system of claim 12, wherein the common processor is operable to output one or more of the following:

an indication of a fluorescence intensity measured by each flow cytometry unit, an indication of sperm cell staining variations, an indication of a rate at which each flow cytometry unit is separating sperm cells, and an indication of a decision boundary used by each flow cytometry unit for discriminating between sperm cells.

15. The multi-channel system of claim 12, wherein the common processor is operable to perform at least one of the following:

(1) receive and process information in real time; and

(2) receive and process information to permit evaluation of the operation of one flow cytometry unit in the plurality of flow cytometry units relative to another flow cytometry unit in the plurality of flow cytometry units.

16. The multi-channel system of claim 1, wherein the processor is operable to receive the digitized output of the analog to digital converter as a substantially continuous stream and to process the digitized output in real time.

17. The multi-channel system of claim 1, wherein said processor is operable to process the digitized output as multiple digital samples in the form of a pulse waveform.

18. The multi-channel system of claim 1, wherein the processor comprises multiple processors.