Method for staining and sorting of a small volume of sperm
AssigneeGenetics & IVF Institute
InventorSayed Bahawaddin HASHEMI
Life of this patent
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Abstract
The present invention relates to a method of sorting of a small volume or low concentration stained sperm sample, the method includes: (a) diluting the stained sperm cells in a ratio of 1:1 or less by volume with a dilution buffer to obtain a diluted volume of said stained sperm sample; (b) placing the diluted volume of the stained sperm sample on a flow cytometer to initiate the sorting process; (c) sorting the stained sperm cells in the diluted volume based upon a preselected sperm cell or stain characteristic; and (d) collecting sorted sperm cells.
Claims17
A method of sorting sperm cells, comprising:
- (a) obtaining a sperm sample comprising a plurality of sperm cells in a volume of 0.3 ml or less or wherein the plurality of sperm cells is less than about 2×106 cells/ml;
- (b) staining the sperm cells by forming a staining mixture comprising intact viable sperm cells and a DNA selective fluorescent dye for a period of time sufficient to allow the dye to bind to DNA of the sperm cells;
- (c) diluting the stained sperm cells of step (b) to obtain a diluted volume of said stained sperm cells;
- (d) sorting the stained sperm cells in the diluted volume based upon a preselected sperm cell characteristic;
- and (e) collecting sorted sperm cells.
A method of sorting a stained sperm sample of less than about 0.3 mL or less than 1×106 sperm cells/mL, comprising:
- (a) diluting the stained sperm sample in a ratio of 1:1 or less by volume with a dilution buffer to obtain a diluted volume of said stained sperm sample;
- (b) subjecting the diluted volume of the stained sperm sample to a flow cytometric sorting process;
- (c) sorting the stained sperm cells in the diluted volume based upon a preselected sperm cell or stain characteristic;
- and (d) collecting sorted sperm cells.
Dependencies are inferred from the claim text.