High pressure sperm sorting and flow cytometer methods
AssigneeInguran, LLC
InventorsKenneth Michael Evans, Thomas Boyd Gilligan, Richard W. Lenz, Ramakrishnan Vishwanath, Clara Gonzalez-Marin
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Abstract
Cell sorting methods that improve sorting efficiency and productivity by elevating sorting pressures and incorporate certain steps to help the cells better survive such elevated pressures. In the case of sperm, sorting the steps of standardizing sperm samples, staining sperm samples in a single step, calibrating a flow cytometer to place sperm in the leading edge of droplets, and changing a catch fluid distance may be incorporated individually, or in combination to help sperm better survive the sex sorting process.
Claims14
A method of sorting bovine sperm comprising:
- extending a bovine sperm sample in an initial extender at a sperm sample to initial extender ratio between about 1:1 and 1:10 to form an extended bovine sperm sample;
- reconcentrating the extended bovine sperm sample to a concentration between 900 million sperm per ml and 2400 million sperm per ml, wherein the reconcentrated sperm concentration is higher than the sperm concentration following dilution;
- staining sperm in the reconcentrated bovine sperm sample with a DNA selective dye;
- and
- sorting the stained sperm with a flow cytometer with a sheath fluid pressure between about 45 psi and about 65 psi, wherein the steps of extending and reconcentrating the bovine sperm sample reduces damage imposed on the sperm by the sheath fluid pressure.
Dependencies are inferred from the claim text.