Systems and methods for sorting particles

• US 8,709,817 B2
• Filed: 02/07/2013
• Issued: 04/29/2014
• 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:
  US13/762,014
• Publication Number:
  US 20130224727A1
• Time in Patent Office: 446 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, 435/286.1, 435/286.5, 435/287.1, 435/288.7, 435/29, 435/366
• US Class Current: 436/63
• 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 flow cytometry system and method for sorting a mixture of particles, such as a fluid switching sorting system for sorting selected particles which includes a control having a microprocessor programmed with instructions for implementing a control strategy for controlling the fluid delivery to the system to vary the rate at which fluid is delivered as a function of the purity or yield of a sub-population of particles.

Claims

1. A flow cytometry system for sorting a mixture of particles including particles having a characteristic A and particles having a characteristic B, the system including:

a fluid delivery system for delivering a fluid containing the particles to a flow cytometry apparatus;

a flow cytometry apparatus for receiving the fluid, forming the fluid into a stream and using flow cytometry to classify the particles according to the characteristics; and

a fluid switching sorting system for sorting selected particles in the stream according to the classification and according to a sorting strategy to provide at least one population containing desired particles, wherein the fluid switching system comprises;

a control having a microprocessor programmed with instructions for implementing a control strategy responsive to information received from the flow cytometry apparatus during sorting for controlling the fluid delivery system during sorting to vary a rate at which fluid is delivered as a function of at least one of the following;

(1) a purity of the said at least one population with respect to either characteristic A particles or characteristic B particles; and

(2) a quantity of characteristic A particles or characteristic B particles in the said at least one population relative to a total quantity of characteristic A particles or characteristic B particles in the stream.

2. The system as claimed in claim 1, wherein the particle are sperm cells, and wherein characteristic A is indicative of a live X sperm cell and characteristic B is indicative of something other than a live X cell (˜

X).

3. The system as claimed in claim 1,wherein the control is suitable for determining the purity based on output signals from the flow cytometry apparatus, the system further comprises an operator input to the control for indicating a desired purity, and wherein the control varies the rate of fluid delivery to obtain the desired purity.

4. The system as claimed in claim 1, wherein the particles within the stream arrive in a series at the flow cytometry apparatus which includes sequential sets of particles, including:

first particle sets each including one or more particles having characteristic A;

second particle sets each including one or more particles having characteristic B; and

third particle sets each including two or more closely spaced particles at least one of which has characteristic A and at least one of which has characteristic B.

5. The system as claimed in claim 4, wherein the fluid switching sorting system diverts either only the second particle sets or the second particle sets and the third particle sets in the stream.

6. The system as claimed in claim 4, wherein the fluid switching sorting system diverts only the second particle sets and the control maintains the purity of the said at least one population at more than 85% but less than 95%.

7. The system as claimed in claim 4, wherein the fluid switching sorting system diverts the second particle sets and the third particle sets in the stream and wherein the control increases the rate of fluid delivery when the quantity of characteristic A particles in the said at least one population is greater than an acceptable quantity of characteristic A particles in the said at least one population, and wherein the control decreases the rate of fluid delivery when the quantity of characteristic A particles in the said at least one population is less than the acceptable quantity.

8. The system as claimed in claim 4, wherein the fluid switching sorting system diverts the second particle sets and the third particle sets in the stream and wherein the control determines the quantity of characteristic A particles in the said at least one population based on output signals from the flow cytometry apparatus, the system further including an operator input to the control for indicating an acceptable quantity of characteristic A particles in the said at least one population, and wherein the control varies the fluid delivery rate to obtain the acceptable quantity of characteristic A particles in the said at least one population.

9. A method of sorting a mixture of particles including particles having a characteristic A and particles having a characteristic B comprising:

providing a flow cytometry system;

delivering a fluid containing the particles to a flow cytometry apparatus;

forming the fluid into a stream and using flow cytometry to classify the particles in the stream according to the characteristics;

sorting the particles in the stream by diverting selected particles in the stream according to the classification and according to a sorting strategy thereby to provide at least one population containing desired particles; and

during sorting varying a rate at which fluid is delivered as a function of at least one of the following;

(1) a purity of the said at least one population with respect to either characteristic A particles or characteristic B particles; and

(2) a quantity of characteristic A particles or characteristic B particles in the said at least one population relative to a total quantity of characteristic A particles or characteristic B particles in the stream.

10. The method as claimed in claim 9, wherein the particles are sperm cells, and wherein characteristic A is indicative of a live X sperm cell.

11. The method as claimed in claim 9, further comprising the step of increasing the rate of fluid delivery when the purity of the said at least one population is greater than a desired purity and decreasing the rate of fluid delivery when the purity is less than the desired purity.

12. The method as claimed in claim 9, wherein the fluid stream contains particles following one after another in a series including sequential sets of particles, wherein the sequential sets of particles comprise:

first particle sets each including one or more particles having characteristic A;

second particle sets each including one or more particles having characteristic B; and

third particle sets each including two or more closely spaced particles at least one of which has characteristic A and at least one of which has characteristic B.

13. The method as claimed in claim 12, further comprising the step of diverting either the second particles sets and not the first and third particle sets or the second and third particle sets and not the first particle sets.

14. The method as claimed in claim 12, further comprising the step of diverting the second particles sets and not the first and third particle sets and further includes maintaining the purity of the said at least one population at more than 85% but less than 95%.

15. The method as claimed in claim 12, further comprising the step of diverting the second and third particle sets and not the first particle sets and further includes increasing the rate of fluid delivery when the quantity of characteristic A particles in the said at least one population is greater than an acceptable quantity of characteristic A particles in the said at least one population, and decreasing the rate of fluid delivery when the quantity of characteristic A particles in the said at least one population is less than the acceptable quantity.