Zebrafish sperm cells

To confirm the technique for sperm concentration measurement with flow cytometry with simultaneous analysis of sperm plasma membrane integrity in zebrafish.
Conclusion:
This study demonstrated that flow cytometry can be used for determination of sperm concentration while simultaneously analyzing membrane integrity. This approach is comparable to counting results obtained from other methodologies such as hemocytometer or absorbance measurement by microspectrophotometer, and the precision and accuracy were retained across a concentration range of 1 × 103 cells ml-1 to 1 × 107 cells ml-1. The approach developed can be used for determination of sperm concentration and quality of valuable small samples. Quality control includes: 1) the peristaltic pump tubing was less than 2 months old; 2) the fluidics of the flow cytometer was calibrated for precise volume measurements by using the same volume and type of sample tube; 3) the correct collection of fluorescent signals by the flow cytometer was validated by use of standard 8-peak and 6-peak beads (from BD Biosciences) before analyzing experimental samples; 4) All experiments had replicates in this study.