COMPARISON OF MANUAL MICROSCOPIC AND COMPUTER-ASSISTED METHODS FOR ANALYSIS OF SPERM COUNT AND MOTILITY

This investigation was conducted to determine which of three methods, manual analysis, and two different commercially available computer-ass isted semen analyzers (CASA), was the most reproducible. Semen samples from donors participating in an artificial insemination program (n = 1) and from patients being seen for andrology procedures (n = 12) were acquired at 0.5 h after ejaculation. Each specimen was loaded into on e chamber of a 20-mu m microcell slide (Conception Technologies, San D iego, CA, USA) and the port was sealed with petroleum jelly to prevent drying of the specimen. The specimens were assessed for sperm count ( SC) and motility (MOT) first by manual analysis using an eyepiece reti cle and brightfield light microscopy at 400x total magnification, seco nd using the Hamilton-Thorn 2030 analyzer (Hamilton-Thom Research, Dan vers, MA, USA), and third, using the Cell Trak/S system (CTS; Motion A nalysis Corporation, Santa Rosa, CA, USA). Each analysis was repeated five times for each specimen on the same microcell by the same technic ian. The three methods were compared in terms of means and standard de viations of the SC and MOT over repeated measures of a specimen using sign tests. The CTS system measured significantly lower sperm counts t han the HTM system. MAN was intermediate and not significantly differe nt from either. For MOT, there were no significant differences. Compar ison of the standard deviations demonstrated that the three methods we re not equally reproducible. For SC, the manual method was significant ly less reproducible than the HTM system; the CTS system was intermedi ate. For MOT, the manual method was less reproducible than either CASA system, both of which were not significantly different from each othe r. CASA methodology in general provides a more reproducible (less vari able) analysis than the manual microscopic method for assessing sperm count and motility.