7. Gently remove the epididymis from the Petri dish using a fine
forceps, and gently swirl the petri dish in order for evenly
distributing the spermatozoa throughout the freezing solution
(see Fig. 3c).
8. Transfer 10 μL sperm in a microcentrifuge tube containing
90 μL FHM to determine sperm concentration and motility
values using preferably Hamilton-Thorne sperm motility analyzer or subjective analysis by an experienced person (see
Note 6).
3.6 Sperm Freezing
1. Using a 1 cc syringe, load approximately 200 μL of sperm
suspension in the freezing solution to five pre-labeled straws
(see Fig. 4), and load ~100 μL into a sixth straw to determine
post-thaw motility later.
Fig. 3 Cauda epididymis are in the freezing solution (a), just after mincing them in freezing solution (b), 10 min
after mincing and their removal (c)
Fig. 2 An adult male mouse reproductive organs (arrowhead indicates cauda epididymis and arrow shows vas
deferens)
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409
forceps, and gently swirl the petri dish in order for evenly
distributing the spermatozoa throughout the freezing solution
(see Fig. 3c).
8. Transfer 10 μL sperm in a microcentrifuge tube containing
90 μL FHM to determine sperm concentration and motility
values using preferably Hamilton-Thorne sperm motility analyzer or subjective analysis by an experienced person (see
Note 6).
3.6 Sperm Freezing
1. Using a 1 cc syringe, load approximately 200 μL of sperm
suspension in the freezing solution to five pre-labeled straws
(see Fig. 4), and load ~100 μL into a sixth straw to determine
post-thaw motility later.
Fig. 3 Cauda epididymis are in the freezing solution (a), just after mincing them in freezing solution (b), 10 min
after mincing and their removal (c)
Fig. 2 An adult male mouse reproductive organs (arrowhead indicates cauda epididymis and arrow shows vas
deferens)
Mouse Sperm Cryopreservation
409