Fluorescent Y in sperm heads after sperm penetration

FLUORESCENT Y IN SPERM HEADS AFTER SPERM PENETRATION. I . Salonen and H. 3unnila. Department of Biomedicine, University of Turku, 20520 Turku, Finland Enrichment of Y-bearing human spermatozoa has been v e r i f i e d by the fluorescent Y (Ybody) in the sperm heads and by sperm karyotyping a f t e r penetration i n t o the zona-free hamster eggs. We w i l l introduce an a l t e r n a t i v e method to i d e n t i f y the Y-spermatozoa in the zona-free hamster egg t e s t . Zons-free hamster ova are a i r - d r i e d on slide 3 hours a f t e r incubation with capacirated human spermatozoa, Fixed in methanol and stained by d i l u t e quinacrine mustard (0.0035 %in water). Swollen sperm heads are visualized in a fluorescence microscope; swollen Y-sperm heads are easily detectable. The frequency of fluorescent Y in the decondensed sperm heeds is approximately 33 % (3 samples, 332 swollen sperm heads). Scoring of fluorescent human Y-bodies in hamster eggs may be used to monitor the success of any Y-sperm enrichment procedure. The advantage of the method is that it simultaneously verifies the apparent fertilizing capacity of the manipulated sperms. Additonally, the present method seems to be valuable in comparison of the frequency of Y-chromosome aneuploidy.

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