we used Egg yolk citrate for semen dilution 7% glycerol and gentamycine. the process is good until transport the semen from refrigerator 4C to the liquid nitrogen vapor for 10 min.
few straws had a good motile semen while other straws (majority) showing dead or abnormal motility
How can we overcome that.
Salam and very good day
There are several impacts which lead to reduce the quality of frozen-thawed semen.
According to references, the period of cooling for extended semen (before cryopreservation) should be at least three hours. Another factor that could influence the quality of semen is the cryopreservation process which should not be very fast (makes ice crystal) and not slow (makes dehydration in sperm cell). The distance between liquid nitrogen and straws during cryopreservation process is also very important and it affects from male to another.
Thus, it recommended for you
1- chilling of extended semen before cryopreservation should be three hours or more.
2- You should try different distances during vapor of straws in liquid nitrogen with different periods (for me I am using 13 min).
3- You should not add the glycerol at chilled period, you should add the glycerol before 5 min of cryopreservation process at chilled temperature.
I hope these comments are useful for you
Regards
This link of video is very useful
https://youtu.be/x747Hs1cwVE
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