Method for wheat tube vitrescence-freezing storaging geneine porgy

A vitrification freezing and preservation method technology, applied in biochemical equipment and methods, preservation of human or animal bodies, microorganisms, etc., can solve the problems of large size and difficult embryo cryopreservation, achieve small size, overcome large-scale instruments The effect of restraint and thawing operation is simple

Inactive Publication Date: 2006-03-22
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
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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
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Embodiment Construction

[0020] The scope of protection of the present invention is not limited to the following examples.

[0021] (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);

[0022] (2) Use 80-mesh sieve silk net to fish out the embryos, lightly soak the surface seawater with filter paper, put it into antifreeze, and the antifreeze is a mixture of various antifreezes, containing 20.5% of dimethyl sulfoxide (volume content) + acetamide 15.5% (volume content) + propylene glycol 10% (volume content) + polyethylene glycol 6% (mass volume ratio, W / V, containing 6g polyethylene glycol in every 100ml solution), balanced salt solution Cortland was used as the base solution (i.e. the balance, pH=8.28), and was quickly inhaled into a 0.5ml ...

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Abstract

The present invention belongs to the field of marine biotechnology, and is technology of obtaining normal revived genuine porgy embryo including vitrification freezing of embryo, long period maintenance in liquid nitrogen, deforesting, eluting, etc. The genuine porgy embryo in palpitation stage is treated through adding antifreezing agent, fast straw vitrification freezing, maintenance in liquid nitrogen, deforesting, eluting and other steps to obtain the revived genuine porgy embryo. The present invention is significant in the germ plasm maintenance, genetic diversity, breeding, etc of genuine porgy.

Description

technical field [0001] The invention belongs to the technical field of marine organisms, and obtains normal revived embryos through vitrification freezing of red sea bream embryos, long-term storage in liquid nitrogen, 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...

Claims

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Application Information

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IPC IPC(8): C12N5/08A01N1/02
Inventor 丁福红李军肖志忠
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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