Microfluidic system and method with focused energy apparatus

Patent metadata

Field	Value
Publication number	US-11415503-B2
Application number	US-202117496469-A
Country	US
Kind code	B2
Filing date	Oct 7, 2021
Priority date	Oct 30, 2013
Publication date	Aug 16, 2022
Grant date	Aug 16, 2022

Abstract

A method of inseminating an animal including flowing a stream of a population of sperm cells through a channel, differentiating the sperm cells into two subpopulations of X-chromosome containing sperm cells and Y-chromosome containing sperm cells, selecting a desired subpopulation, ablating an undesired subpopulation, and collecting both the subpopulations of sperm cells including the desired subpopulation and the ablated undesired subpopulation together, wherein the collected population of sperm cells is used to fertilize an egg.

First claim

What is claimed is:

1. A method for producing a sexed composition of sperm comprising non-human animal sperm by identifying a subpopulation of sperm from a population of sperm in a sample fluid, the method comprising:
staining the population of sperm in the sample fluid with a dye;
defining a gating strategy for differentiating the subpopulation of sperm in the sample fluid in the population of sperm based on one or more physical characteristics of sperm in the population of sperm, wherein the one or more physical characteristics are identifiable within the population of sperm based in part on the dye staining, and wherein the gating strategy comprises identifying one or more desired physical characteristics of the subpopulation of sperm;
measuring the one or more physical characteristics for the sperm in the population of sperm;
measuring a span distribution of the sample fluid based on the defined gating strategy;
identifying sperm in the subpopulation of sperm based on the measured one or more physical characteristics and the defined gating strategy; and
generating the sexed composition of sperm comprising the subpopulation of sperm identified based on the measured one or more physical characteristics, the span distribution, and the defined gating strategy.

2. The method of claim 1 , wherein the population of sperm in the sample fluid comprises bovine sperm.

3. The method of claim 1 , wherein the sexed composition of sperm substantially comprises X chromosome-bearing sperm cells.

4. The method of claim 1 , wherein the sexed composition of sperm substantially comprises Y chromosome bearing sperm cells.

5. The method of claim 1 , wherein the dye is Hoechst 33342.

6. The method of claim 1 , wherein the gating strategy further comprises identifying the one or more desired physical characteristics of the subpopulation of sperm in a plot of the one or more physical characteristics of the sperm in the population of sperm.

7. The method of claim 6 , wherein the gating strategy further comprises plotting a location of each physical characteristic of the one or more physical characteristics for each sperm cell in the population of sperm in the sample fluid in a graphic representation.

8. The method of claim 7 , wherein the gating strategy further comprises marking the subpopulation of sperm in the graphic representation of the one or more physical characteristics.

9. The method of claim 7 , wherein the graphic representation is a histogram.

10. The method of claim 7 , wherein the method further comprises determining whether an acceptable resolution of the one or more physical characteristics of the sperm is present in the plot.

11. The method of claim 1 , wherein a physical characteristic of the one or more physical characteristics is an amount of DNA.

12. A method for defining a gating strategy for use in producing a sexed composition of sperm comprising non-human animal sperm by identifying a subpopulation of sperm from a population of sperm in a sample fluid, the method comprising:
determining one or more desired physical or genetic characteristics to be present in the sexed composition of sperm;
setting gating parameters for differentiating the subpopulation of sperm in the sample fluid in the population of sperm based on the one or more physical or genetic characteristics of sperm in the population of sperm resulting in a defined gating strategy, wherein the one or more physical or genetic characteristics are identifiable within the population of sperm based in part on a dye staining of the population of sperm in the sample fluid with a dye, and wherein the gating parameters comprise marking the one or more desired physical or genetic characteristics of the subpopulation of sperm;
measuring the one or more physical or genetic characteristics for the sperm in the population of sperm;
measuring a span distribution of the sample fluid based on the defined gating strategy;
identifying sperm in the subpopulation of sperm based on the measured one or more physical or genetic characteristics and the defined gating strategy; and
generating the sexed composition of sperm comprising the subpopulation of sperm identified based on the measured one or more physical or genetic characteristics, the span distribution, and the defined gating strategy.

13. The method of claim 12 , wherein the population of sperm in the sample fluid comprises bovine sperm.

14. The method of claim 12 , wherein the sexed composition of sperm substantially comprises X chromosome bearing sperm cells.

15. The method of claim 12 , wherein the sexed composition of sperm substantially comprises Y chromosome bearing sperm cells.

16. The method of claim 12 , wherein the dye is Hoechst 33342.

17. The method of claim 12 , wherein the marking of the one or more desired physical or genetic characteristics of the subpopulation of sperm further comprises marking the one or more desired physical or genetic characteristics of the subpopulation of sperm in a graphic representation of the one or more physical or genetic characteristics of the sperm in the population of sperm.

18. The method of claim 17 , wherein the gating parameters further comprise a plot of a location of each physical or genetic characteristic of the one or more physical or genetic characteristics for each sperm cell in the population of sperm in the sample fluid in the graphic representation.

19. The method of claim 18 , wherein the graphic representation is a histogram.

20. The method of claim 12 , wherein the method further comprises determining whether an acceptable resolution of the one or more physical or genetic characteristics of the sperm is present in the graphic representation.

21. The method of claim 12 , wherein a physical or genetic characteristic of the one or more physical or genetic characteristics is an amount of DNA.

22. A method for producing a sexed composition of sperm comprising non-human animal sperm by identifying a subpopulation of sperm from a population of sperm in a sample fluid, the method comprising:
staining the population of sperm in the sample fluid with a dye;
defining a gating strategy for differentiating the subpopulation of sperm in the sample fluid in the population of sperm based on one or more physical characteristics of sperm in the population of sperm, wherein the one or more physical characteristics are identifiable within the population of sperm based in part on the dye staining, and wherein the gating strategy comprises identifying one or more desired physical characteristics of the subpopulation of sperm in a plot of the one or more physical characteristics of the sperm in the population of sperm;
measuring the one or more physical characteristics for the sperm in the population of sperm;
measuring a span distribution of the sample fluid based on the defined gating strategy;
identifying sperm in the subpopulation of sperm based on the measured one or more physical characteristics and the defined gating strategy; and
generating the sexed composition of sperm comprising the subpopulation of sperm identified based on the measured one or more physical characteristics, the span distribution, and the defined gating strategy.

23. A method for defining a gating strategy for use in producing a sexed composition of sperm comprising non-human animal sperm by identifying a subpopulation of sperm from a population of sperm in a sample fluid, the method comprising:
determining one or more desired phy

Assignees

• Abs Global Inc

Inventors

• Appleyard David
• Betthauser Jeff
• Faust Marjorie
• Larsen John
• Shao Guocheng
• Xia Zheng
• Zhou Yu

Classifications

• B01L2300/0887

  Laminated structure · CPC title

• G01N21/01

  Arrangements or apparatus for facilitating the optical investigation · CPC title

• G01N2015/1413

  Hydrodynamic focussing · CPC title

• G01N33/4833 Primary

  of solid biological material, e.g. tissue samples, cell cultures (tissue in vivo A61B5/00; cell suspensions G01N33/48735) · CPC title

• G01N15/1484 Primary

  microstructural devices · CPC title