Semen freezing and preservation
Abstract: In order to fully utilize and improve the utilization rate of elite rams and improve the quality of frozen and thawed semen, the enzyme kinetic spectrophotometry was used to study the changes of ATPase activity in spermatozoa and spermatozoa of small-tailed Han sheep before and after cryopreservation.
Freezing and preservation of semen: The well-balanced semen straw is placed on a self-made frozen floater (2.0 to 2.5 cm from the surface of liquid nitrogen, and the temperature is at -110 to -80 °C) for 10 minutes, and stored in liquid nitrogen.
Thawing of semen: Remove the semen straw from the liquid nitrogen tank, quickly immerse it in a water bath of 39 to 40 ° C, and gently shake until it melts (10 to 15 s).
Evaluation of sperm viability: After shaking the diluted semen, take 10 μL of the middle semen and drop it on the isothermal preheated glass slide at 37 °C. Cover the sheet and place it under the microscope to enlarge 400 times and check the sperm for more than 1000 pieces. The number of spermatozoa in the linear motion and the total number of spermatozoa were calculated to calculate the sperm motility.
During cryopreservation, sperm undergoes a process of interaction with cryogens, cooling, cryopreservation, and thawing and rewarming. The physical and chemical effects (pH, osmotic pressure, ice crystal formation, etc.) in these processes are It may damage sperm and cause ATPase activity to be affected.
Cryopreservation may cause damage to sperm structure and function.
Because: 1. The formation of ice crystals during the freezing process and the change of the osmotic pressure inside and outside the cell lead to the damage of the sperm membrane. The lipoprotein in the sperm membrane is separated from the cell structure, which changes the original spatial conformation of the cell membrane, and the enzyme in the sperm cell. Released into the refined clear; 2. Endogenous sperm lipid peroxidation can cause serious damage to the sperm membrane, leading to the release of cell contents, exogenous fatty acid peroxide can also damage the sperm membrane, resulting in the same intracellular The release of the enzyme is consistent with the results of Chen Tianfei et al.
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