Application of liquid nitrogen tank in biological experiment

1. Use in cell culture process

The general process of cell culture mainly includes four steps of preparation, material extraction, culture and cryopreservation. In order to preserve cells, particularly mutant cells or cell lines that are not readily available, the cells are cryopreserved. The cryopreservation temperature is generally at a temperature of 196 ° C liquid nitrogen, the cells are collected into a cryotube and added with a medium containing a protective agent (generally dimethyl sulfoxide or glycerol), frozen at a certain cooling rate, and finally preserved. In liquid nitrogen. At very low temperatures, the time it takes for cells to be preserved is almost limitless.

At present, the US standard cell bank or cell bank (ATCC) liquid nitrogen has 3200 identified cell lines (1992), including skin fibroblast cell lines from normal people and various disease patients and near from different species. 75 hybridoma cell lines.

2. Embryo cryopreservation technology

Embryo cryopreservation technology refers to a technique for preserving embryos using cryoprotectants under low temperature conditions. This technology was produced in the second half of the 20th century with the extensive application of artificial insemination technology, and has become one of the necessary means of modern life science research. The principle of cryopreservation of animal embryos is because the low temperature reduces the rate of metamorphic reaction in the embryo. The lower the temperature, the longer the embryo preservation time.


 

A variety of embryo cryopreservation methods have been developed, including slow freezing, rapid freezing, one-step freezing, direct-transfer freezing, and vitrification. The steps of the vitrification method are as follows:

1 The embryo was pretreated in a 25% vitrification solution (VS solution) at room temperature for 15 min;

2 The pre-treated embryos are stored in a high concentration (75% - 90%) of VS solution for 20-40 min;

3 The embryos treated with the two-step antifreeze are directly stored in liquid nitrogen.

In addition, liquid nitrogen tanks can be used to preserve strains, vaccines, bone tissue, sperm and other substances, which can achieve good long-term storage effects.

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