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can i use 50ml tube contain isopropanol alchol


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#1 semsem shaltout

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Posted 26 February 2014 - 09:49 AM

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



#2 jerryshelly1

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Posted 26 February 2014 - 10:16 AM

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.



#3 semsem shaltout

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Posted 26 February 2014 - 10:32 AM

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



#4 bob1

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Posted 26 February 2014 - 12:24 PM

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.



#5 CellApplicationsInc

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Posted 26 February 2014 - 02:25 PM

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.






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