Freezing cells in DMSO straight into minus 80?
Posted 18 January 2012 - 07:38 PM
Posted 19 January 2012 - 01:03 AM
I tried and did it here, and when I thawed my cells they were very fine (According to my little experience)
Posted 19 January 2012 - 02:44 AM
No, If anyone does it, they have less chance to recover the cells.
If you donot have Isopropanol gradual freezing system. One can use Thermocol Box. Keep or insert the vial in to thermocol box in a way the atleast the liquid part goes into the thermocol, some time i use to do it when i donot have enough space in gradual freezing system.
Edited by Biouday, 19 January 2012 - 02:45 AM.
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Posted 19 January 2012 - 02:52 AM
Dear science noob,
You cannot freeze cells straight into a -80 freezer. Generally these days people use a "Mr. Frosty (£50) or a controlled rate freezer machine (£15-20K). I have my own method which costs next to nothing:-
Cells in DMSO/Glycerol into ice for 10 minutes
Then put the cells in a polystyrene box, with a lid. Perforate the box with the tip of a small pair of scissors. Fill the box with cheap tissue paper and put the vials into it......then put the box into a -20 freezer for 2 hours.
After 2 hours put the box into a -80 freezer and leave overnight.
Next day transfer the frozen vials into a liquid nitrogen repository.
Always works for me....usually 90% viability when next initiated into a TC flask.
Hope this is useful
Posted 19 January 2012 - 12:49 PM