4. Place a coverslip on the hemocytometer after wetting supports
(rails) with saliva or water. This helps to hold down the coverslip
while loading the sperm.
5. Place 10-15 µl of the diluted sperm under the cover slip
on each side of the hemocytometer as shown in the figure
6. Carefully place the hemocytometer in the prewetted chamber,
close the lid and wait 5 minutes.
7. Remove the hemocytometer without tilting it, dry the bottom
if needed and place on a microscope. View the hemocytometer with
a 40 X objective.
8. If you have used a phase type hemocytometer (the thin ones)
then be sure the phase rings are in and the condensor is all the
way up. You should be able to focus on the sperm but be careful
not to break the coverslip. If you used one of the thicker hemocytometers,
pull the phase rings out and close down the apeture on the condensor
so that you can see the sperm. You may also need to raise or lower
the condensor so you can see the sperm.