Method agreement between three different chambers for comparative bull semen computer assisted sperm motility analysis

Abstract

The aim of this study was to compare computer assisted sperm analysis (CASA) results of frozen thawed bull semen using three different chambers. Sixty bull frozen semen samples were thawed (37°C; 30 sec), extended in PBS (30×106 spermatozoa/mL; 37°C) and incubated (37°C; 2 min). Each semen sample was analyzed by CASA [total motility, progressive (pro)/ non-progressive/rapid/medium/slow movement spermatozoa, VCL, VSL, VAP, ALH, BCF, LIN, STR, WOB and hyperactive spermatozoa] using three different chambers: a Makler® chamber (MC; 10 μm); a Leja 4 chamber slide (LC; 20 μm); and a Glass slide covered with a coverslip (GSC; 10.3 μm). The Makler chamber gave higher values compared to both the LC and GSC for almost all examined parameters. No systematic effect was evident between LC and GSC for VCL, VSL, VAP, LIN, STR, WOB, ALH, and BCF. Method agreement between MC and LC was generally moderate, between MC and GSC poor and between LC and GSC moderate to good. In general, narrower limits of agreement were found in samples with lower values. In conclusion, the CASA outcomes could be influenced by the analysis chambers. This finding should be taken into consideration when comparing results from different laboratories.