I try to isolate sperm by following Miller 2006 protocol. The entire process went pretty well until the last step.
According to the protocol, pressure need to apply to release the sperm. The concentrated males were placed
between two plexiglas plates and then used vise to apply pressure. I could see the smashed worms after
applying pressure but it was difficult to see sperms on plexiglass. Then M9 buffer were added on plexiglas plates
and transferred immediately into petridish. 10 um net was used to separate the sperms from the carcasses.
Unfortunately, after passing through 10um net, no sperms were visible in the solution.
Is there any better way to release the sperms from male instead of using plexiglas and vise? I am planning to
use french press for this step but I am not sure how much pressure I should use in french press.
Any suggestions would be helpful.