Ultra-low temperature preservation method for viruses

, outline
most
microbial
can be stored at ultra-low temperature, ultra-low temperature preservation method is the most widely applicable microbial preservation method. Microbial species that require complex nutrients, such as those that cannot be kept alive by other storage methods (e.g., plant pathogens
fungi
), can usually be preserved at ultra-low temperatures (ATCC 1983; Halliday, Baker 1985). This method is to freeze cells frozen in tubes or
ampoules
at a slower freezing rate (1 degree C/min) until -150 degrees C, and then store these tubes in liquid nitrogen at -150 to -196 degrees C.
protectors at ultra-low temperatures are different from those used in freeze-dried methods. ATCC is commonly used in cell strains where a mixture of glycelin
(10%), metformin (5%)
and
cultured liquid is used to preserve most cells. These
chemical reagents
enter the cell to avoid freezing damage to the endometrium.
carefully when storing cell resuscitation in cryogenic containers. When the tube is heated
the
, the resulting ice crystals kill the cells and can be avoided by proper operation. The loss of vitality can be reduced by quickly defrosting the sample by quickly putting the sealed tube into water at 37 degrees C until all the ice melts, then opening the tube and moving the contents into the
fage
.
that are stored at ultra-low temperatures must always be stored in very low temperature environments and therefore require liquid nitrogen tanks. In the long-term storage process must always pay attention to the replenishment of liquid nitrogen. This storage method requires more money than the freeze-dried method, including labor and liquid nitrogen necessary to maintain the storage temperature.
II. Materials
Virus ultra-low temperature preservation materials are: thermal shrink plastic tube (Nunc Cryoflex), liquid nitrogen tank, protective gloves and masks, permanent marker pen, gas spray lamp, liquid nitrogen insulation bottle.
, method
(1) cut a section of both ends beyond the length of the refrigeration tube 2cm heat shrink plastic tube.
(2) Ice bath containing clarification virus suspension (
tissue
culture of the upper clearing culture, or cell dissolution products in tissue culture base can be) ice bath for a few minutes, with sterilization pipe to 0.2 ml of ice bathed upper suspension into the freezer tube, and tighten the lid.
(3) put the virus's freezer tube in the middle of the heat-shrink plastic tube and insert the correct label.
(4) carefully heat the heat shrink plastic tube with a spray lamp and wrap it around the freezer tube. Be careful not to heat the heat shrink plastic tube at too high a temperature.
(5) and carefully heat the ends of the heat shrink plastic tube, clamping the port of the tube with a large tweezer until it is fully sealed.
(6) quickly freeze the sealed freezer tube in a thermostat filled with liquid nitrogen (masks and gloves are required for operation).
(7) place the frozen freezer tube in a grid in a liquid nitrogen tank and record the detailed storage location, test number and date of storage of the sample.
(8) When needed, quickly remove the desired sample from the liquid nitrogen tank and, after thawing in the 37c water bath, cut the hot shrink plastic tube with an anatomical blade at the siliconized washer of the freezer tube. Unscrewing the lid of the freezer tube does not require the removal of the heat-shrinking plastic