can i use 50ml tube contain isopropanol alchol - (Feb/26/2014 )

hi:all

i haven"t the container of  isopropanol alchol that placed in  -80, can i simply use 50ml tube filled with that alchol and put the cryovial inside it then in -80??

second..there is about 13 min between putting demso for freezing my  lymphocytes and to reach -80 freezer how can transport the cells without lost in these minutes? and what is the importance of one hour in  -20 before -80?

kind regard

asmaa

I wouldn't put the cryovial into direct contact with the isopropanol. A good substitute would be to put the cryovial in the old Styrofoam that was used to house 15mL tubes.

I never put my lymphocytes in the -20C before placing them in the -80C.

jerryshelly1 on Wed Feb 26 18:16:10 2014 said:

I wouldn't put the cryovial into direct contact with the isopropanol. A good substitute would be to put the cryovial in the old Styrofoam that was used to house 15mL tubes.

 

I never put my lymphocytes in the -20C before placing them in the -80C.

thanks for answers but how can transport the cells in freezing media before reach -80 for 13 min

Just transport them on ice, most cells will tolerate DMSO for a short period of time.  You could also substitute glycerol for DMSO, which is less toxic but more tricky to use due to its osmotic strength.

the reason for -20C before -80C is that you want to gradually freeze your cells.

Without any special containers, you can first keep the cryovials on ice for ~20 minutes, then move them to -20C, then put them in a container filled with dry ice and ethanol and transport them in that container. The temperature of dry ice+ethanol is supposed to be close to -80. BTW, after a day or two at -80 I would transfer the frozen cells to liquid nitrogen for any long term storage.  If you keep them at -80 for longer than a few months, their viability will drop significantly.

Hope this helps.