Methods for improving sheath fluids and collection systems for sex-specific cytometer sorting of sperm
DC CAFCFirst Claim
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1. A method of sorting cells by flow cytometry comprising the steps of:
- a. establishing a cell source which supplies cells to be sorted;
b. chemically coordinating a sheath fluid to create a sheath fluid environment for said cells which is coordinated with both a pre-sort and a post-sort cell fluid environment and combining said cells with said sheath fluid;
c. sensing a property of said cells in a flow cytometer;
d. discriminating between cells having a desired characteristic; and
e. collecting cells having said desired characteristic.
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Abstract
Improved flow cytometer system particularly adapted to use for sex-selected sperm sorting include enhanced sheath fluid and other strategies which minimize stress on the sperm cells, including a 2.9 percent sodium citrate sheath solution for bovine species and a HEPES bovine gamete media for equine species. Improved collection systems and techniques for the process are described so that commercial applications of sperms samples as well as the resulting animals may be achieved.
180 Citations
45 Claims
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1. A method of sorting cells by flow cytometry comprising the steps of:
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a. establishing a cell source which supplies cells to be sorted;
b. chemically coordinating a sheath fluid to create a sheath fluid environment for said cells which is coordinated with both a pre-sort and a post-sort cell fluid environment and combining said cells with said sheath fluid;
c. sensing a property of said cells in a flow cytometer;
d. discriminating between cells having a desired characteristic; and
e. collecting cells having said desired characteristic. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 41, 42, 43)
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32. A method of sorting cells by flow cytometry comprising the steps of:
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a. establishing a cell source which supplies cells to be sorted;
b. establishing a sheath fluid to create a sheath fluid environment for said cells and combining said sheath fluid with said cells;
c. sensing a property of said cells in a flow cytometer;
d. discriminating between cells having a desired characteristic;
e. collecting cells having the desired characteristic in a collector fluid; and
f. chemically coordinating said collector fluid with said sheath fluid to create an ending collector fluid environment for said cells which is coordinated with a pre-sort fluid environment. - View Dependent Claims (33, 34, 35, 36, 37, 38, 39, 40, 44, 45)
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Specification