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Invisibleagar
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Can anyone verify-->
    #6440547 - 01/09/07 01:42 AM (17 years, 4 months ago)

Spores can be frozen without significant loss of viability by suspending in a 10% aqueous solution of sterilized glycerol (glycerin U.S.P.; available at most pharmacies)


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Invisiblefastfred
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Re: Can anyone verify--> [Re: agar]
    #6440564 - 01/09/07 01:55 AM (17 years, 4 months ago)

Yes that's true. Probably depends a bit on what you consider "significant loss of viability" though.

"Genetics and Breeding of Edible Mushrooms" has some good info on this if I remember right. There are lots of preservation methods and variables like freezing rate.


-FF

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InvisibleSillicybin
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Re: Can anyone verify--> [Re: fastfred]
    #6440839 - 01/09/07 07:46 AM (17 years, 4 months ago)

Found this after a bit of research....

Quote:

Ice crystals, however, are lethal to living cells. [...] the secret is to add a substance like glycerol that prevents the formation of ice crystals.




Once site I read suggests 50% glycerol by volume, though?

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OfflineSchwip
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Re: Can anyone verify--> [Re: agar]
    #6447843 - 01/11/07 07:29 AM (17 years, 4 months ago)

is glycerol the stuff diabetics get in those little vacutainers?


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Offlineshirley knott
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Re: Can anyone verify--> [Re: Schwip]
    #6448456 - 01/11/07 11:17 AM (17 years, 4 months ago)

er.... that would be insulin


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Offlinefunkyjunky
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Re: Can anyone verify--> [Re: shirley knott]
    #6457947 - 01/14/07 08:59 AM (17 years, 4 months ago)

He's probably thinking of Glucose, maybe glycogen?

Is the purpose of the glycerin/glycerol to seal the spores from water or oxygen? Would a 100% dessicated spore print in an oxygen free jar have the same shelf-life as a glycerol suspension, if both were in the freezer?

Agar, I wish I could find the source, but I thought it was you who quoted a respected mycology author on the long term glycerol storage? Maybe I'm confusing that with the vacutainers..


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Offlinecaricapapaya
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Re: Can anyone verify--> [Re: funkyjunky]
    #6461867 - 01/15/07 11:31 AM (17 years, 4 months ago)

I work growing massive amounts of e. coli, and we store all of our cultures in a medium with 25% glycerol (glycerin).

we store them at -80 C.

carica

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Invisibleagar
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Re: Can anyone verify--> [Re: caricapapaya]
    #6464458 - 01/15/07 11:54 PM (17 years, 4 months ago)

Interesting job, you must have. :wink:
Thanks for the info.


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Invisiblephalcon005

Registered: 12/21/05
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Re: Can anyone verify--> [Re: agar]
    #6485020 - 01/22/07 08:49 AM (17 years, 3 months ago)

I've worked with several organisms and stored at -80C in a glycerol mix. Human breast cancer cells were mixed in liquid culture with an equal part of glycerol. Lyme disease and other Borrelia spirochetes are mixed with 33% glycerol and usually less than half die in the entire process. The glycerol prevents the little cells from lysing due to the freezing temps, but the catch I believe is that you want to get them away from glycerol when you want the grow them again. At first I thought of simply adding spores to 100% glycerol, but then you will have a sticky mess that you'll probably have to dilute out and plate to get to grow again.

It seems that lyophilizing the spores (freeze drying) might be the best bet if one could do it. Where I work we have bacterial cultures that were lyophilized as far back as the 40's and they are still viable after being stored at -20C all those years.

That's my two cents...

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