Liquid Nitogen + Cell Culture in Glycerol = Frozen Cell Stock

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Uploaded by on Jun 30, 2010

Preparation of frozen cell stock using liquid nitrogen. The low boiling point of nitrogen helps rapidly freezing the cell culture + glycerol for storage in -80 degrees Celsius. Rapidly freezing the culture and the mixing it with glycerol at a 1:1 ratio (for example, 0.75ml cell culture + 0.75ml glycerol) is required in reducing cell lysis (which causes cell death) due to the shape of the ice crystals that can pierce the cell membrane. If either liquid nitrogen or the glycerol is not used the frozen stock will be less viable at best during the cell revival process. - mypetridish

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Liquid nitrogen is nitrogen in a liquid state at a very low temperature. At atmospheric pressure, liquid nitrogen boils at −196 °C, and is a cryogenic fluid which can cause rapid freezing on contact with living tissue, which may lead to frostbite.

Liquid nitrogen is a compact and readily transported source of nitrogen gas without pressurization. Further, its ability to maintain temperatures far below the freezing point of water makes it extremely useful in a wide range of applications, primarily as an open-cycle refrigerant such as the coolant for CCD cameras in astronomy, to store cells at low temperature for laboratory work, and in cryogenics.

- WIkipedia entry on Liquid Nitrogen

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