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Method for freezing embryo of fish

A freezing method and embryo technology, applied in the preservation, application, and animal husbandry of human or animal bodies, can solve problems such as ultrastructural damage, frozen embryos cannot survive, and embryo death due to toxic effects, so as to reduce damage, eliminate and Reduce the damage of ice crystals to embryos, and the effect of strong experimental repeatability

Inactive Publication Date: 2005-03-30
EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

[0002] The existing technology uses low-concentration antifreeze to freeze and store fish embryos. The balance treatment time before freezing is 10 minutes. Because the excess water in the cryopreservation process cannot be fully removed, ice crystals are easy to form and cause direct damage to the embryos. Can not survive; adopt slow cooling and segmented slow cooling in the cooling rate, and cannot quickly cross the dangerous temperature zone (0~-60°C) that is easy to freeze, which will easily cause the water in the embryo to freeze, and the ice crystals will damage the nucleus of the cells. The ultrastructure of cell organelles such as mitochondria and mitochondria are severely damaged. Due to the slow cooling, the embryos spend too long in the antifreeze. Although part of the water can be removed, the toxic effect of the antifreeze also causes the death of the embryos. Freezing still doesn't work

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Embodiment Construction

[0007] The best way to implement the present invention is to use the easy vitrification solution combined with two or more high-concentration antifreeze agents as the antifreeze protection agent for cryopreservation of fish embryos; process flow in the flow chart 1. represents vitrification solution Prepared with 1.2 propylene glycol + DMSO or 1.2 propylene glycol + methanol; use a solution prepared with a concentration of 25% to 30% of 1.2 propylene glycol and a concentration of 10% to 15% DMSO or a concentration of 20% to 30% of 1.2 propylene glycol and a concentration of 10% to The solution prepared with 15% methanol has low toxicity, no hyperosmotic damage, and is easy to vitrify; for example, the vitrification solution formula for preserving loach embryos is 1.2 propylene glycol 25%+DMS015%, that is, the total is 60ml distilled water+25ml1. 2 propylene glycol + 15ml DMSO; different types of fish embryos have different proportions for cryopreservation and vitrification, but...

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Abstract

A method for freeze storage of fish embryo without damage to it includes such steps as preparing the antifreeze protective liquid from two or more antifreezing agents temp balancing loading the mixture in centrifugal tube, quickly putting it in liquefied nitrogen at -196 deg.C for storage, and thawing which includes temp lalancing at -180 deg.C and thawing by water bath at 38-40 deg.C.

Description

Technical field: [0001] The invention relates to a method for cryopreserving fish embryos in liquid nitrogen. Background technique [0002] The existing technology uses low-concentration antifreeze to freeze and store fish embryos. The balance treatment time before freezing is 10 minutes. Because the excess water in the cryopreservation process cannot be fully removed, ice crystals are easy to form and cause direct damage to the embryos. Can not survive; adopt slow cooling and segmented slow cooling in the cooling rate, and cannot quickly cross the dangerous temperature zone (0~-60°C) that is easy to freeze, which will easily cause the water in the embryo to freeze, and the ice crystals will damage the nucleus of the cells. The ultrastructure of cell organelles such as mitochondria and mitochondria are severely damaged. Due to the slow cooling, the embryos spend too long in the antifreeze. Although part of the water can be removed, the toxic effect of the antifreeze also cau...

Claims

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Application Information

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IPC IPC(8): A01N1/02
Inventor 章龙珍庄平张涛冯广朋黄晓荣
Owner EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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