Study objectives: To develop methods of instrument calibration and standardization that enable the use of computer-aided sperm analysis (CASA) technology in multicenter studies of sperm motility.
Setting: Clinical semen evaluation for the andrology laboratory and in vitro fertilization programs.
Patients: Semen specimens were selected from the videotape archives of the University of California at Davis Andrology Laboratory and used to produce a videotape reference standard for CASA instrument testing and calibration. Four identical CASA instruments, two each at two sites, were used to analyze the videotape. Machine accuracy was verified by manual analysis of the videotape.
Results: The coefficient of variation (CV) for repeated measures was between 1% and 8% for each variable on all CASA instruments. Mean values for CASA parameters varied as a function of the digitization threshold. A minimum CV was obtained at a particular threshold setting, suggesting an optimum for each machine. Because of high within-machine precision, slight differences in mean values between machines were statistically significant for some variables and some specimens; such differences are probably not biologically significant.
Conclusion: Multicenter standardization of CASA instrumentation is possible, but threshold settings are one source of variation that must be controlled using CVs or another means of objective calibration. One method is to adjust the threshold of each instrument until the count obtained by CASA equals the count obtained by the manual method used to determine the dilution requirements for neat semen.