Vitrifiation freezing preservation method for genine porgy embryo particles

A technology of cryopreservation and granulated glass, which is applied in animal husbandry and other directions, can solve the problems of large volume and difficulty in cryopreservation of embryos, and achieve the effect of small volume, favorable for rapid freezing and large heat dissipation area.

Inactive Publication Date: 2006-03-22
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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Problems solved by technology

Due to the characteristics of fish embryos, especially seawater fish embryos, which are large i...

Method used

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Embodiment Construction

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

[0024] (1) Take the same batch of naturally-produced red sea bream fertilized eggs and place them in a slightly inflated incubator with a sea temperature of 18±1°C. The embryos at the heartbeat stage will have more than 60 beats per minute (60-140 beats per minute);

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

[0026] (3) Take the embryos with an 80-mesh silk screen, lightly dip filter paper to absorb the surface seawater, put it in a 20% glycerol volume concentration antifreeze, and use a balanced salt solution Cortland as the base fluid (i.e. balance, pH=8.28), Soak for 15 minutes, then transfer to 40% glycerol volume concentration antifreeze, balance salt solution Cortland as base liquid (i.e. balance, pH=8.28), and quic...

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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 present invention belongs to the technical field of marine biology, and specifically describes the process of obtaining resurrected embryos by freezing and thawing and eluting the red sea bream embryos. Background technique [0002] The 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, types and toxicity of antifreeze, and have made great progress. At present, cryopreservation of carp 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. Acta Fisheries 1997, 21 ( 4): 366-372), salmon and trout embryos also broke the -25°C limit (Erdahl, DA, Preservation of spermatozoa and ova from freshwater fish. Thesis of the U...

Claims

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Application Information

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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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