A simple and reproducible method for separating Y-bearing spermatozoa from human semen.

ABSTRACT

PIP A simple and reproducible method for separating Y-bearing spermatozoa from human semen is reported. Semen samples with 60-80 million count and 50% motility were obtained from healthy human donors. Ficoll-400 and sodium metrizoate density gradient were used for separation. Quinacrine hydrochloride was used for staining, and Olympus fluorescent microscopy was used for the detection of F body present in the spermatozoa bearing Y-chromosomes. The data indicate that the percentage of Y-bearing spermatozoa in normal human semen is about 5-6% less than what is expected. The observations agree with those of Barlow and Vosa that with the optical attenuation coefficient being sufficiently great, the chromosomes lying deep in the head may go undetected. A mean value of 45.22 was obtained for Y-bearing spermatozoa and the sediment contained about 62.9 Y-bearing spermatozoa. The results thus suggest that it is possible to separate Y-bearing spermatozoa from semen using Ficoll-socium metrizoate density gradient.^ieng