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Abstract
This investigation was conducted to determine which of three methods, manual analysis, and two different commercially available computer-assisted 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 one chamber of a 20-μm microcell slide (Conception Technologies, San Diego, 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 reticle and brightfield light microscopy at 400x total magnification, second using the Hamilton-Thorn 2030 analyzer (Hamilton-Thorn Research, Danvers, MA, USA), and third, using the Cell Trak/S system (CTS; Motion Analysis Corporation, Santa Rosa, CA, USA). Each analysis was repeated five times for each specimen on the same microcell by the same technician. The three methods were compared in terms of means and standard deviations of the SC and MOT over repeated measures of a specimen using sign tests. The CTS system measured significantly lower sperm counts than the HTM system. MAN was intermediate and not significantly different from either. For MOT, there were no significant differences. Comparison of the standard deviations demonstrated that the three methods were not equally reproducible. For SC, the manual method was significantly less reproducible than the HTM system; the CTS system was intermediate. For MOT, the manual method was less reproducible than either CASA system, both of which were not significantly different from each other. CASA methodology in general provides a more reproducible (less variable) analysis than the manual microscopic method for assessing sperm count and motility.