Use of a Giemsa stain to detect changes in acrosomes of frozen ram spermatozoa

Acrosomal structures of ram spermatozoa were prominently stained when air dried smears of diluted semen were fixed for 15 minutes in buffered formal saline and stained for 90 minutes in a 6 per cent (v/v) buffered solution of Giemsa stain. Progressive disruption of the acrosomes was demonstrated during chilling and deep-freezing of the spermatozoa, and the degree of damage was systematically scored. A rapid and repeatable estimate of the state of the acrosomes in a sample could be made from the mean score of 20 spermatozoa examined per slide.