Comparison of commercially available chamber slides for computer-aided analysis of human sperm

Abstract

Despite the increasing use of computer-aided sperm analysis (CASA) in clinical practice, there is still no golden standard for the type of slide to be used with these systems. Counting chamber depth and loading method, can profoundly influence motility and concentration estimates, thereby undermining the validity and accuracy of CASA. To contribute toward standardized sperm analysis, this study compared different commercially available capillary-filled slides including 10 and 20 µm deep Leja slides (Leja10 and Leja20); 10, 16 and 20 µm deep CellVision slides (CV10, CV16 and CV20); and drop-loaded slides including slide and coverslip (SCS) with a depth of 20.1 µm and the Makler chamber with a depth of 10 µm for sperm analysis when using CASA. The Sperm Class Analyzer (SCA) CASA system was used to assess concentration, motility, and detailed kinematic parameters of 20 normozoospermic human samples using the different chamber slides. Results were evaluated by the repeated measures ANOVA and Intraclass correlation coefficients. The Makler chamber showed significantly (P < 0.05) higher concentrations than other slides. However, there was no significant difference in the percentage of sperm in different motility groups among the slides. CV10, Leja10 and Makler showed significantly higher curvilinear-, average path- and straight-line velocity (VCL, VAP, VSL) values than other slides. In conclusion, despite the objectiveness of the assessments by CASA systems, there are still some discrepancies in the results of sperm concentration, motility and other kinematic parameters when using different commercially available slides. The possible negative influence of the sperm quality misdiagnosis on the selection of treatment strategy in a clinical setting, emphasizes the need for further standardization and quality control of the commercially available chamber slides for use with CASA. Furthermore, this study found more consistent results for capillary-filled chambers compared to drop-loaded slides, suggesting a superior method when using CASA.Abbreviations: DNA: Deoxyribonucleic acid; CASA: Computer-aided semen analysis; SCA: Sperm Class Analyzer; WHO: World Health Organization; CV: CellVision; SCS: Slide and Coverslip; SD: Standard deviation; ANOVA: Analysis of Variance; PR: Progressive; NP: Non-progressive; IMT: Immotile; VCL: Curvilinear velocity; VAP: Average path velocity; VSL: Straight-line velocity; Lin: Linearity; STR: Straightness; WOB: Wobble; ALH: Amplitude of lateral head displacement; BCF: Beat cross frequency.

Keywords: CASA; Chamber slide; Concentration; Motility; Sperm quality.