Photo-damage method for sorting particles

• US 8,206,987 B2
• Filed: 05/12/2011
• Issued: 06/26/2012
• Est. Priority Date: 03/28/2003
• Status: Expired due to Term

• 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:
  US13/106,671
• Publication Number:
  US 20110217722A1
• Time in Patent Office: 411 Days
• Field of Search: 422/67, 422/68.1, 422/73, 422/82.05, 422/82.08, 436/63, 436/164, 436/165, 436/43, 436/50, 436/55, 436/172, 435/2, 435/287.1, 435/286.1, 435/286.5, 435/366, 435/29, 435/288.7
• 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 system and method for sorting a mixture of stained particles in a fluid flow path, including stained particles from two distinguishable groups. The system and method can include an electromagnetic radiation source for exciting fluorescence emissions from the stained particles, a photodetector for detecting the fluorescence emissions from the stained particles, a processor for classifying the stained particles; and a photo-damaging laser for damaging selected particles in the flow path.

Claims

1. A method of sorting a mixture of stained sperm cells having either characteristic A or characteristic B into at least one population, the method comprising the steps of:

a. flowing a fluid stream containing stained sperm cells through a flow path at a fluid delivery rate;

b. exciting fluorescence emissions from the stained sperm cells having characteristic A and the stained sperm cells having characteristic B flowing in the flow path;

c. detecting the fluorescence emissions from the excited sperm cells;

d. classifying the stained sperm cells as either having characteristic A or having characteristic B based upon the fluorescence emissions;

e. selecting stained sperm cells in the flow path based on their classification; and

f. photo-damaging the selected sperm cells to produce an enriched population of sperm with respect to either characteristic A or characteristic B.

2. A method of sorting a mixture of stained sperm cells according to claim 1 wherein the step of selecting stained sperm cells in the flow path to photo-damage further comprises the step of photo-damaging sperm cells based upon a sort strategy.

3. A method of sorting a mixture of stained sperm cells according to claim 2 further comprising the step of varying the sort strategy as a function of at least one of the following:

a. the purity of the said at least one population with respect to either characteristic A sperm cells or characteristic B sperm cells; and

b. the quantity of undamaged characteristic A sperm cells or undamaged characteristic B sperm cells in the said at least one population relative to the total quantity of characteristic A sperm cells or characteristic B sperm cells in the fluid stream.

4. A method of sorting a mixture of stained sperm cells according to claim 2 further comprising the step of varying the fluid delivery rate as a function of at least one of the following:

a. the purity of the said at least one population with respect to either characteristic A sperm cells or characteristic B sperm cells; and

b. the quantity of undamaged characteristic A sperm cells or undamaged characteristic B sperm cells in the said at least one population relative to the total quantity of characteristic A sperm cells or characteristic B sperm cells in the fluid stream.

5. A method of sorting a mixture of stained sperm cells according to claim 1, wherein the method further includes the step of forming the fluid into a stream containing sperm cells following generally one after another in a series which includes sequential sets of sperm cells, including:

a. first particle sets each including one or more sperm cells having characteristic A;

b. second particle sets each including one or more sperm cells having characteristic B; and

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

6. A method of sorting a mixture of stained sperm cells according to claim 5, the method further includes the step of either photo-damaging the second particles sets and not the first and third particle sets or photo-damaging the second and third particle sets and not the first particle sets.

7. A method of sorting a mixture of stained sperm cells according to claim 1 wherein characteristic A is indicative of a live X-chromosome bearing sperm cell (X) and wherein characteristic B is indicative of something other than a live X-chromosome bearing sperm cell (˜

X).

8. A method of sorting a mixture of stained sperm cells according to claim 1, wherein characteristic A is indicative of a live Y-chromosome bearing sperm cell (Y) and wherein characteristic B is indicative of something other than a live Y-chromosome bearing sperm cell (˜

Y).

9. A method of sorting a mixture of stained sperm cells according to claim 1, further comprising the step of increasing the fluid delivery rate 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.

10. A method of sorting a mixture of stained sperm cells according to claim 1 further comprising the step of increasing the fluid delivery rate when the quantity of undamaged characteristic A sperm cells in the said at least one population is greater than an acceptable quantity of undamaged characteristic A sperm cells in the at least one population, and decreasing the rate of fluid delivery when the quantity of undamaged characteristic A sperm cells in the said at least one population is less than the acceptable quantity.