Method for preserving large volume of lefteye flounder sperms by refrigeration
A cryopreservation and large-capacity technology, applied in the field of marine biology, can solve the problems of small storage capacity, low density, and small storage quantity of flounder sperm, and achieve the effects of low toxicity, large volume, and reduced damage
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[0017] The large-capacity cryopreservation method of flounder sperm of the present invention, the steps are as follows:
[0018] (1) Take a cryopreservation tube with a volume of 5ml, collect the semen of flounder, put it into the cryopreservation tube and add antifreeze solution, the volume ratio of semen and antifreeze solution is 1:1. The antifreeze liquid uses Hank’s balanced salt solution as the base liquid, and uses propylene glycol (PG) as the antifreeze agent, and the volume content of propylene glycol in the antifreeze liquid is 15%. The cryopreservation tubes were equilibrated in a refrigerator at 0-4°C for 10 minutes. Take six 5ml cryopreservation tubes from the refrigerator, absorb 5ml of the mixture each, and quickly put them into the programmed cooling device to start cooling.
[0019] (2) The cooling program is: 0~-80°C, the cooling rate is 20°C / min; -80°C balance for 2 minutes; -80~-180°C, the cooling rate is 30°C / min; when it drops to -180°C, Quickly put the ...
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Abstract
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