Patent ID: 9453849
Filing Date: 2016-09-27
CPC Classification: A61D,B82Y,C12N,G01N

Claim Text:
1. A method which distinguishes bovine sperm cells based on DNA content, wherein live sperm cells are stained with a DNA-selective fluorescent dye in an osmotic gradients process, comprising: incubating the live sperm cells in a buffer-containing first solution in a hypertonic condition to partially dehydrate the sperm cells, at a temperature between about 0° C. and about 12° C., which is substantially sufficient to maintain a comparatively high sperm viability rate, and for a time period less than about 5 minutes; transferring the solution containing the sperm cells to a buffer-containing second solution in a hypotonic condition at said same temperature between about 0° C. and about 12° C.; wherein the hypertonic condition is about more than 250 mOsm and the hypotonic condition is about less than 250 mOsm, for bovine sperm cells; wherein the dye is present in at least one of the hypertonic or hypotonic condition to expedite permeation of the dye into the sperm cells when osmotic equilibrium is reached, thereby staining DNA in the sperm cells; exposing the sperm cells to a light source to cause the stained DNA to fluoresce; detecting a pre-determined fluorescence of the stained DNA, the predetermined fluorescence corresponding to DNA content; distinguishing the sperm cells based on the pre-determined fluorescence; and collecting selected sperm cells.