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

Inactive Publication Date: 2010-05-19
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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Problems solved by technology

[0004] The object of the present invention is to provide a large-capacity cryopreservation method for flounder sperm, which

Method used

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Embodiment

[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

The invention belongs to the technical field of marine organisms and particularly relates to a method for preserving large volume of lefteye flounder sperms by refrigeration. The method comprises the following processes: refrigerating large volume of lefteye flounder sperms; preserving the lefteye flounder sperms by liquid nitrogen; and unfreezing and eluting the preserved lefteye flounder sperms to obtain the sperms which have fertilization ability. The method comprises the following steps of: collecting lefteye flounder semen; placing the semen and anti-frozen liquid to a 5ml of freezing tube in a volume ratio of 1:1; pre-cooling the semen and the anti-frozen liquid at 0 to 4 DEG C for 9 to 11 minutes; lowering the temperature to -180 DEG C in section by section and placing the lefteye flounder semen in the liquid nitrogen for storing; and directly placing the lefteye flounder semen to water at a temperature of between 40 and 42 DEG C to melt the lefteye flounder semen and unfreezing the molten lefteye flounder semen at room temperature so as to obtain sperms with the fertilization ability. The method for preserving large volume of lefteye flounder sperms by refrigeration plays an important role in breeding, germplasm conservation, genetic diversity, sustainable aquaculture and the like for of rare seawater fishes such as lefteye flounder, and solves the problems of low preservation volume, low density, small preservation quantity and the like in prior art.

Description

technical field [0001] The invention belongs to the technical field of marine organisms, and specifically relates to a large-capacity cryopreservation method for flounder sperm, which is a process of freezing large-capacity flounder sperm, storing them in liquid nitrogen, thawing and eluting them, and obtaining sperm with fertilization ability. Background technique [0002] Flounder is an important marine economic fish in my country. It is not only an important fishing target in my country, but also a major species of marine aquaculture. Flounder is distributed in the western part of the North Pacific Ocean and is endemic to Northeast Asia on the west coast of the Pacific Ocean. It is mainly distributed in China's Bohai Sea, Yellow Sea, East China Sea, South China Sea, and the coastal waters of the Korean Peninsula, Japan, and Russia's Far East. Over the past 30 years, due to overfishing and increasingly serious environmental pollution, the natural resources of flounder have...

Claims

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Application Information

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IPC IPC(8): A01N1/02
Inventor 刘清华肖志忠李军
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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