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Vitrifiation freezing preservation method for genine porgy embryo particles

A particle glass, cryopreservation technology, applied in animal husbandry and other directions, can solve the problems of large size and difficulty in embryo cryopreservation, and achieve the effect of small size, conducive to rapid freezing and reducing toxicity

Inactive Publication Date: 2007-11-14
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

Due to the characteristics of fish embryos, especially seawater fish embryos, which are large in size and contain a large amount of yolk and water, it is very difficult for embryo cryopreservation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0023] The scope of protection of the present invention is not limited to the following examples.

[0024] (1) Pick up fertilized eggs of the same batch of naturally produced red sea bream, place them in a slightly inflated hatching bucket with a seawater temperature of 18±1°C, and cultivate them until the embryos are in the heartbeat stage, and the heartbeat frequency is more than 60 times / minute (60-140 times / minute);

[0025] (2) Put a certain amount of liquid nitrogen into the incubator, the liquid level is about 10cm high, put the copper foil (thickness 12 microns) flat on the liquid nitrogen surface and fix it.

[0026] (3) Grab the embryos with 80 mesh sieve silk nets, lightly soak the surface seawater with filter paper, put it into the antifreeze agent with a volume concentration of 20% glycerin, and use the balanced salt solution Cortland as the base liquid (i.e. the balance, pH=8.28), Soak for 15 minutes, then transfer to antifreeze with 40% glycerin volume concentra...

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PUM

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Abstract

The present invention belongs to the field of marine biological technology, and especially the process of granular freezing genuine porgy embryo, preservation in liquid nitrogen, de-freezing and eluting to obtain live embryo. The genuine porgy embryo in cardiopalmus stage is added with antifreeze agent and fast granular vitrification frozen and then treated through preservation in liquid nitrogen, de-freezing and eluting, so as to obtain live embryo. The present invention is significant in germ plasm preservation, genetic diversity, breeding and cultivation of genuine porgy.

Description

technical field [0001] The invention belongs to the technical field of marine biology, and specifically relates to the process of obtaining revived sea bream embryos through granule freezing, liquid nitrogen storage, thawing and elution. Background technique [0002] Research on cryopreservation of fish embryos has a history of about 20 years. Scholars at home and abroad have done a lot of research on the damage mechanism of fish embryo cryopreservation, freezing and cooling rate, thawing and rewarming, antifreeze types and toxicity, and have made great progress. At present, the cryopreservation of Cyprinidae embryos at temperatures above -100°C has achieved a relatively stable resurrection rate (Zhang Kejian et al. Research on cryopreservation of three freshwater fish embryos and their cooling and rewarming rates. Journal of Fisheries, 1997, 21( 4): 366-372), salmon and trout embryos also broke through the limit of -25°C (Erdahl, D.A., Preservation of spermatozoa and ova f...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01K67/033
Inventor 丁福红李军肖志忠
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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