The human acrosome, especially the acrosome reaction, is an important parameter for evaluation of sperm function. By histochemical staining, the acrosome is visualized at the light microscopic level. However, a high number of cells are lost due to the multiple centrifugation steps that are required for preparation of spermatozoa. In the present study, semen samples from oligozoospermic patients were stained by an improved method. Compared to the conventional triple-stain technique, optimal staining of the acrosome and a lower number of cells lost during sperm preparation were observed (P<0.025). Differences in the percentage of live acrosome-reacted spermatozoa were not statistically significant.