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Simple method for ultralow temperature preservation of fish sperm

A technology for ultra-low temperature freezing and fish sperm, which is used in the preservation of human or animal bodies, household refrigeration devices, lighting and heating equipment, etc. It can solve the problems of low sperm count, inconvenience, sperm death, etc. The effect of cryopreservation and easy operation

Inactive Publication Date: 2007-12-19
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this freezing method is that the amount of sperm preserved each time is very small, and a suitable thawing solution is required for thawing, so few people use it.
The collection of rare fish sperm often takes several years or even decades to have a chance to collect sperm. Once missed, it is difficult to collect again
However, the field is too far away from the laboratory, and sometimes it takes 1-2 days to return to the laboratory. In this way, the collected sperm are all dead when they return to the laboratory.
And the program cooling instrument or the computer control program cooling instrument are bulky, very heavy, and it is not convenient to move to the field for work.

Method used

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  • Simple method for ultralow temperature preservation of fish sperm

Examples

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Effect test

Embodiment 1

[0020] In the figure, there is a box cover 1 on the top of the foam box 2, there is liquid nitrogen 4 in the foam box 2, and there is a foam board 3 on the top of the liquid nitrogen 4, and the foam board 3 is a hollow board with a total thickness of 25 mm and a length of 25 cm , and the width is 100 millimeters, and 9 wheat straws 5 are placed on the foam board 3 .

Embodiment 2

[0022] The following is an example of freezing 10ml carp sperm to illustrate the specific operation process of the simple freezing method:

[0023] (1) After collecting 10ml of carp sperm with fresh semen, place it in a refrigerator at 4°C, take 5ml of fresh semen each time, add 20ml of diluent to a 30ml vial bottle and mix well, the total volume is 25ml , equilibrate in a refrigerator at a temperature of 4°C for 30 minutes, the diluent is 8 grams of NaCl, 0.5 grams of KCl, 15 grams of glucose, distilled water to 1000 ml, pH6.0;

[0024] (2) After the balance is completed, add 2.16 milliliters of DMSO (dimethyl sulfone antifreeze) to make the concentration of the antifreeze reach 8%. All disassembly can be completed, the shorter the disassembly time, the better, usually only 30-60 seconds;

[0025] (3) Place the packed straws horizontally on the foam board on the liquid nitrogen surface of the foam box. When the hollow width of the foam board is 10 cm, the length is 20 cm, an...

Embodiment 3

[0029] The specific operation process of the freezing method is illustrated below by freezing 20 ml of mullet sperm:

[0030] (1) After the fresh essence is collected, put it in a refrigerator at 4°C, take 10ml of fresh essence each time, add 30ml of diluent to a 50ml vial and shake well, the total volume is 40ml, and place it in a refrigerator at 4°C Equilibrate for 30 minutes, and dilute it with 8 grams of NaCl, 0.5 grams of KCl, 15 grams of glucose, and distilled water to 1000 ml, pH7.0;

[0031] (2) After the balance is completed, add 3.26 milliliters of DMSO (dimethyl sulfone antifreeze agent) to make the concentration of the antifreeze agent reach 8%. All sub-assembly can be completed, the shorter the sub-assembly time, the better, usually only 30-60 seconds;

[0032] (3) Place the packed straws horizontally on the foam board on the liquid nitrogen surface of the foam box. When the hollow width of the foam board is 10 cm, the length is 30 cm, and the thickness is 3 cm, ...

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Abstract

The simple ultralow temperature method of preserving fish sperm includes the following steps: taking fish sperm, diluting with diluent in 1-5 times, balancing in a refrigerator at 4 deg.c for 20-30 min, adding antifreeze agent in certain concentration, packing in 0.5 mm diameter straw, setting on foamed plastic board over liquid inside a foamed plastic case for 20 min, setting the straw into liquid nitrogen inside the case for 5 min, and transferring into liquid nitrogen container for freeze preservation at the temperature of -196 deg.c. The present invention may be used in field operation of preserving fish sperm, and is fast, simple, high in freeze preserving effect and low in cost.

Description

Technical field: [0001] The invention relates to a method for cryopreserving fish sperm at ultra-low temperature Background technique [0002] At present, there are three methods for ultra-low temperature cryopreservation of fish sperm: one is the pellet freezing method, the other is the ampoule freezing method, and the third is the straw freezing method. The pellet freezing method is to directly drop fish sperm into dry ice or freeze them into small sperm particles with liquid nitrogen vapor, then transfer them to a liquid nitrogen tank, and use a program cooling device or a computer-controlled program cooling device to control the freezing of fish sperm Speed, so that the fish sperm freezing temperature reaches -196 ℃ for preservation. The disadvantage of this freezing method is that the amount of sperm preserved each time is very small, and a suitable thawing solution is required for thawing, so few people use it. The ampoule freezing method is to put the fish sperm int...

Claims

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Application Information

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IPC IPC(8): A61K35/52A01N1/00F25D3/10
Inventor 柳凌张洁明危起伟郭峰朱永久
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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