Semen analysis results obtained by a recently developed semi-automated Autosperm system (Amsaten N.V.S.A. Corp., De Pinte, Belgium) were compared with those obtained by the conventional, time-exposure photomicrographic, and automated CellSoft system (Cryo Resources Inc., New York, NY) analyses. The Autosperm system either over- or underestimated the sperm concentration in comparison with the conventional analysis and more often underestimated the sperm concentration in comparison with the automated CellSoft system analysis. Comparison of the results for percent sperm motility by the conventional and Autosperm analyses showed that the latter tended to underestimate the percentage of fast-swimming spermatozoa and overestimate the percentage of slow-swimming spermatozoa. There were considerable variations in the measurement of sperm movement characteristics between the Autosperm and time-exposure photomicrographic and automated CellSoft system analyses, respectively. These findings demonstrate that the performance of the Autosperm system does not agree well with those of the currently available methods employed in the present study. Part of the disagreement in measurements of sperm parameters could be because of the subjective elements inherent in the semi-automated Autosperm analysis.