Determination of sperm concentration and viability for artificial insemination
OwnerPreben Christensen, Jens Peter Stenvanginventors
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OwnerPreben Christensen, Jens Peter Stenvanginventors
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A method for the simultaneous determination of the total concentration of sperm cells and the proportion of live sperm cells in a semen sample is provided. By selective staining of live, dead and/or dying sperm cells, for example using one or more fluorochromes, the detection means can distinguish between live, dead and/or dying sperm cells, for example by detection of the fluorescent response from the sperm cells. The selective staining may be performed at room temperature and in concentrations below standard concentrations. The determination may be performed by selective counting of cells of each fluorescent quality for example by using a flow cytometer having internal concentration standard means. Furthermore, the likelihood of fertilizing a female animal with an insemination dose, taken from a semen sample having a known total concentration of sperm cells and a known proportion of live sperm cells, is predicted on the basis of the thus determined total concentration of sperm cells and the proportion of live sperm cells.
A method for the determination of the absolute concentration of sperm cells in a semen sample and the proportion of live sperm cells therein, comprising subjecting the semen sample to selective staining, wherein said selective staining comprises applying a first fluorochrome which binds to DNA and stains all sperm cells and a second fluorochrome which binds to DNA and selectively stains dead and dying cells, and determining the absolute concentration of the sperm cells and the proportion of live sperm cells by means of a detection means responsive to the selective staining, wherein the determination of the absolute concentration of sperm cells and of the proportion of live sperm cells in the semen sample are performed simultaneously in the same determination step.
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