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Offlineragadinks
MrBeatle
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Registered: 10/20/03
Posts: 1,298
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Re: Storing mycelium in liquid nitrogen ? [Re: suboriginal]
    #2710195 - 05/21/04 07:08 AM (19 years, 10 months ago)

Somewhere I have read that the mycelium that survived deep freezing was growing much more aggressive and rizomorphic after thawing.
But there were also more deformation of the fruitbodies grown from that mycelium.
Sorry, I cannot remember anymore where I have read that.
If I find the source again I will post a link.

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Offlinepeachy
Transientpsychonaut

Registered: 05/05/04
Posts: 41
Loc: Preserved deep within the...
Last seen: 19 years, 8 months
Re: Storing mycelium in liquid nitrogen ? [Re: ragadinks]
    #2710416 - 05/21/04 09:12 AM (19 years, 10 months ago)

Well all this talk has really piqued my curiousity :smile:
So besides the frozen cake I mentioned earlier, I've ordered myself some agar and am going to try several of the various methods of storing mycelium mentioned in the various links in this thread.

So to recap, I'm intending to create:

1) Batch of 15 test tubes of mycelium stored in plain distilled water at  room temperature in a cupboard. Every 30days, I will use a sterile pipette to remove a small ammount to colonize some agar to test viability & growth.

2) Batch of 15 test tubes of mycelium stored in plain distilled water and frozen chest freezer. Similarly tests will be conducted for viability. Given the limitation of my current stock of test-tubes the maximum duration is probably 15 x 30days as I suspect the temperature change to pipette samples out will destroy or weaken the contents of the tubes... (Yes thats if the first test even succeeds which I doubt.)

3) Batch of 15 glycerine & mycelium mix test tubes stored and tested as above. These will be the interesting ones to watch I think.

4) 1 Frozen and sealed PK cake in a ziplock bag. I will cut off small pieces of mycelium from within the freezer each month to avoid exposing the cake too many temperature alterations. Im thinking of trying to construct some sort of sealed glove box with a drawer for mycelium removal in my freezer for this so that the ambient temperature around the cake would be relatively stable.
I would then try to re-animate the pieces of mycelium in various fashions. (Any suggestions?) so as to minimize damage.

When I have more cakes available in the coming months I might try storing some smaller entire cakes or sections of cakes in glycerine; but for now supply is my limitation :smile:

For the record all of this will be attempted with Ecuadorian mycelium from a strain I've had particularly good results from to date. For comparison every "test" will be fruited to completion to allow comparison of fruit speed, quantity and quality. The final test shall be ingestion by five willing participants in a legal environment :smile:

I realize results from this wont be very conclusive nor scientific in the proper sense as I don't have enough controls & environmental control (wish I had lab access). Regardless they should be noteworthy if combined with existing information. I'll start a new thread to recap this once results become available.

Now if there's someone out there with access to a liquid nitrogen storage facility of some description willing to conduct similar tests?


--------------------
"Time bends, space is boundless. It squashes a man's ego. I feel lonely. Tell me, though... does Man, that marvel of the universe, that glorious paradox who sent me to the stars, still make war against his brother? Keep his neighbor's children starving?"

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Offlinepeachy
Transientpsychonaut

Registered: 05/05/04
Posts: 41
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Re: Storing mycelium in liquid nitrogen ? [Re: peachy]
    #2710494 - 05/21/04 09:48 AM (19 years, 10 months ago)

FYI, here's the cake from which all the mycelium for what I describe above will come. As you can see its a little bit away from full colonization.





Vital stats:

The substrate is whole rice grain mixed in with ground whole rice flour and and 10% ground quinone. The sterile top layer is my vermiculite substitute which is something made out of crushed corn used for reptile cages. The "lid" is tinfoil with with two coffee filters (one inside, one outside) held on by a giant cable twist and some tape for the final filter. The jar was innoculated late night on 10/05. As you can see its quite a sizeable jar :smile:

Little does this poor organism know of what's in store for it :|


--------------------
"Time bends, space is boundless. It squashes a man's ego. I feel lonely. Tell me, though... does Man, that marvel of the universe, that glorious paradox who sent me to the stars, still make war against his brother? Keep his neighbor's children starving?"

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Offlineragadinks
MrBeatle
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Registered: 10/20/03
Posts: 1,298
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Re: Storing mycelium in liquid nitrogen ? [Re: peachy]
    #2712214 - 05/21/04 05:00 PM (19 years, 10 months ago)

Sounds good !
Curious how the results of that experiment will be ...

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Offlinesuboriginal
enthusiast

Registered: 05/06/04
Posts: 278
Last seen: 19 years, 5 months
Re: Storing mycelium in liquid nitrogen ? [Re: ragadinks]
    #2712912 - 05/21/04 08:45 PM (19 years, 10 months ago)

I'm loving the enthusiasm and can hardly wait for the results...


--------------------
Peace, love and organic brown rice...

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Offlinesuboriginal
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Registered: 05/06/04
Posts: 278
Last seen: 19 years, 5 months
Re: Storing mycelium in liquid nitrogen ? [Re: suboriginal]
    #2718866 - 05/23/04 03:08 PM (19 years, 10 months ago)

I don't think the lack of controls is a big problem re 'the fundamental question', that is can mycelia withstand freezing... sounds like you will be able to answer that unequivicanly once and for all, re your strain. The 'speed of fruiting/ morphology of fruits' etc may well differ from what you're used to (and this is where you'd really need to run 'fresh' controls each time to get an answer), but again I don't think that matters too much... I suspect that a few generations later (with no freezing) the more aggressive etc variants would preveil, and the phenotype re growth/ fruiting rates etc will be restored to resemble that of the original strain you froze in the first place. Just guesses, I'm hanging for the actual results...


--------------------
Peace, love and organic brown rice...

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Offlinesuboriginal
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Registered: 05/06/04
Posts: 278
Last seen: 19 years, 5 months
Re: Storing mycelium in liquid nitrogen ? [Re: ragadinks]
    #2721723 - 05/24/04 06:49 AM (19 years, 10 months ago)

Thought I would add this to this thread, it's with regard to P. cubensis, from a thread called 'P.cubensis mycelia may withstand freezing' or sommat like that, posted by a 'chrisdab':

Ive dunked a cake before and had it frozen for aprox. 24 hours. The mycelium survived and after 3 weeks the second flush appeared.
(posted by "chrisdab" in another thread)


--------------------
Peace, love and organic brown rice...

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InvisibleSpeeker

Registered: 02/11/04
Posts: 882
Re: Storing mycelium in liquid nitrogen ? [Re: ragadinks]
    #2748719 - 05/31/04 01:01 AM (19 years, 9 months ago)

CRYOPRESERVATION OF FUNGI
WFCC Technical Information Sheets

other storage related sheets..
FREEZE-DRYING OF MICROORGANISM USING A SIMPLE APPARATUS
LIQUID-DRYING OF MICROORGANISM USING A SIMPLE APPARATUS

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