Patent ID: 7713687

Claim:
A method of using stained sperm cells collected from mammals, comprising the steps of: collecting semen from a male of a non-human mammal; freezing said semen; thawing said semen; incubating sperm cells contained within said semen in a concentration of Hoechst 33342 stain of greater than 40 micro-molar; establishing the temperature at which said sperm cells in said concentration of Hoechst 33342 stain are incubated between about 30 degrees centigrade and about 40 degrees centigrade; adjusting a duration of time said sperm cells are incubated with said concentration of Hoechst 33342 stain; staining DNA within said sperm cells with sufficient uniformity to allow X-chromosome bearing sperm cells to be differentiated from Y-chromosome bearing sperm cells based upon the magnitude of fluorescence; determining the sex characteristic of a plurality of sperm cells contained within said frozen-thawed semen; separating said sperm cells according to the determination of their sex characteristic; isolating sperm cells separated according to the determination of their sex in a collected element; establishing a sample from said sperm cells isolated in said collection element; successfully fertilizing at least one egg of a female non-human mammal with said sample at success levels of at least about 70% of success with spermatozoa that have not been separated and/or frozen; and successfully producing offspring non-human mammal of the desired sex.