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Question about Glycerol freezing

Bacteria Fungi Freezing

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#1 Ziv

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Posted 21 April 2020 - 09:03 PM

Hi all,

 

 

I recently moved into a new lab that has a -80 C freezer, up untill now I used to freeze my cells in a standard -20 C freezer. the freezing  protocol I'm using consists of a 1-step Glycerol:PBS 50:50 freeze.

 

I'm thinking about moving my stocks to the -80 C freezer, and would love to hear thoughts on why it would be better for them. Also, in case I'm deciding to move my currently -20 C frozen tubes into -80 C, is there any intermediate step I should be aware of?

 

My stock currently consists of:

 

E.Coli ATCC/ Listeria Mono ATCC/ Salmonella Thypi/ Candida Albicans ATCC/ Penicilum Rubens ATCC

 

And several more gram negative Bacteria.

 

 

Thanks!

 

 

Ziv

 



#2 mdfenko

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Posted 21 April 2020 - 11:00 PM

Storage at -80C is more stable than at -20C. Degradation is slowed further by the lower temperature.

 

50:50 glycerol should be fine at -80C although the solution will freeze. The glycerol will reduce ice crystal formation so more cells will remain viable when defrosted than without the glycerol.


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#3 Ziv

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Posted 22 April 2020 - 04:40 AM

Thank you! So I can just go ahead and move my tubes to the -80 freezer? no intermediate step?(for example dry Nitrogen treatment) 



#4 mdfenko

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Posted 22 April 2020 - 04:45 PM

You should be able to just transfer them. If, however, you have doubts, then transfer one of each strain to the -80C, holding the rest at -20C during this test. After a few days plate some to compare viability.

 

-80C is generally used for long term storage of aliquots.


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#5 Ziv

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Posted 23 April 2020 - 02:07 AM

Thanks again!







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