Automated sperm concentration analysis with a new flow cytometry-based device, S-FCM.

TLDR: Semen samples from 104 men attending a male infertility clinic were used to evaluate the reproducibility of results and the carryover rate with the S-FCM by performing between- and within-run imprecision analyses, showing that automated analyses were highly reproducible and the Carryover rate was 0.17% or less.

Abstract: The S-FCM uses flow cytometry technology to measure sperm concentrations. Semen samples from 104 men attending a male infertility clinic were used to evaluate the reproducibility of results and the carryover rate with the S-FCM by performing between- and within-run imprecision analyses. In addition, sperm concentrations measured with the S-FCM were compared with those obtained by manual analyses with the Makler chamber and the improved Neubauer hemacytometer. The results showed that automated analyses with the S-FCM were highly reproducible and the carryover rate was 0.17% or less. In within-run imprecision assays, the coefficients of variation for the S-FCM were less than 5% at all sperm concentrations, while those for the Makler chamber were between 17.7% and 28.7% at lower sperm concentrations. The overall correlation between values measured with the S-FCM and those measured with the Makler chamber and improved Neubauer hemacytometer was excellent, but at lower sperm concentrations the correlation was lower. The S-FCM performed sperm concentration analyses in 110 seconds compared with 5 minutes for the Makler chamber and 10 minutes for the improved Neubauer hemacytometer. The S-FCM is suitable for quantitative measurement of lower sperm concentrations. Sperm counts are an essential step in the evaluation of male fertility and usually are performed by microscopic examination of semen by trained personnel. The World Health Organization (WHO) recommends the use of a hemacytometer for determining sperm concentrations in semen, 1 and several methods of using these devices have been described. 1-3 However, the accuracy and reproducibility of analytic data obtained with hemacytometers reportedly are not good enough to compare results from different institutions. 4-6 A number of studies have shown that manual counting is associated with the greatest potential for gross analytic errors in measuring sperm concentrations. 4-8 Consequently, computer-assisted semen analyzers were developed to overcome the subjectivity of sperm counting, but most of these analyzers use some sort of counting chamber or hemacytometer that is itself a source of intrinsic errors. When using a counting chamber or hemacytometer, only spermatozoa located in it can be counted, and only very small quantities of specimens are used to estimate the sperm concentration in 1 mL of semen. Recently we developed a new device (S-FCM) for measuring sperm concentrations that uses flow cytometry technology and an argon laser as the light source. To improve accuracy, the S-FCM device uses 90 times greater quantities of semen than are used with a counting chamber or hemacytometer. In this study, we compared the accuracy and reproducibility of the S-FCM with that of the Makler sperm counting chamber. We also evaluated the feasibility of using the S-FCM in clinical settings by comparing the analytic data with those obtained by using the improved Neubauer hemacytometer or the Makler chamber.