To begin, collect the samples and incubate them for 30 to 60 minutes at room temperature to liquefy completely. Then, mix the sample and load seven microliters into a prewarmed sperm counting chamber at 37 degrees Celsius. Place the chamber on a prewarmed stage of a direct light microscope.
Next, open the computerized sperm analysis software and click on mot to enter the motility and concentration module. Select the configuration that corresponds to human sperm conditions. Randomly analyze 10 different fields per chamber by clicking on the Analyze button.
Then click on Results to obtain the sample concentration and motility. To prepare the cells for high-resolution respirometry, first centrifuge the samples at 400 to 500 G for 10 minutes at room temperature. Then, remove the seminal plasma and resuspend the spermatozoa in two milliliters of appropriate medium.
Repeat the sperm evaluation with computer-aided sperm analysis software for sperm concentration studies.
