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Offlineragadinks
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Storing mycelium in liquid nitrogen ?
    #2696608 - 05/18/04 04:35 PM (12 years, 6 months ago)

In this article at mushworld.com they say that they store mycelium in liquid nitrogen.
Does anybody know more about this kind of long term storage ?


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Offlinepeachy
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Re: Storing mycelium in liquid nitrogen ? [Re: ragadinks]
    #2696681 - 05/18/04 04:43 PM (12 years, 6 months ago)

Sounds interesting, but the site requires registration :frown:

I wonder how cost effective this would be for the small scale (psychotropic) gardener though?

Ideas?


--------------------
"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
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Re: Storing mycelium in liquid nitrogen ? [Re: peachy]
    #2696768 - 05/18/04 04:54 PM (12 years, 6 months ago)

Quote:

Sounds interesting, but the site requires registration



Registration is very easy and does not cost anything.

Quote:

I wonder how cost effective this would be for the small scale (psychotropic) gardener though?
Ideas?




On that site it says:
Quote:

I recommend minimizing transfers by storing in liquid nitrogen, which if you have a non-mechanical tank is not all that expensive or technical.



I wonder what they mean with inexpensive ?


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Offlinepeachy
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Re: Storing mycelium in liquid nitrogen ? [Re: ragadinks]
    #2696870 - 05/18/04 05:07 PM (12 years, 6 months ago)

Yeap, I think I will register and have a gander at that.
Inexpensive is relative, it could mean to your average commercial edible shroom grower; in which case "inexpensive" to them would be synoynmous to "astronomical" for you or I :smile:

I have nasty visions of me (after "accidentally" ingesting something psychedelic) attempting to retrieve some mycelium to innoculate my latest batch- somehow submersing my hand in liquid nitrogen. Imagine the panic! Then tripping and falling over something and my hand  shattering or some shit. LOL! Too much TV eh?


--------------------
"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
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Re: Storing mycelium in liquid nitrogen ? [Re: ragadinks]
    #2697097 - 05/18/04 05:37 PM (12 years, 6 months ago)

Hmm, well the storage (if this is what you'd need) looks  affordable
I suppose my next questions would be:
Where can I find someone unscrupulous enough to fill one such container for me? I suppose extraction of nitrogen in some manner is feasible, but cost-effective? And how long would such a method permit me to store my mycelium-brimming testtube babies?

I suspect the answer to the latter would make it more viable for the smallscale grower just to keep a colonized PF cake in the fridge. A similar type of container states it can hold its contents (180 test tube sized vials) at -150C for 230 days ... That isn't that long if you factor in the lengths you'd have to go to achieve all this in addition to the relatively small capacity of such a container.  Dedicating a suitably large fridge to cake storage sounds waay more sensible I'd have to conclude :smile:

Still, the idea is appealing! :laugh: It'd make for really cold beer too.


--------------------
"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
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Re: Storing mycelium in liquid nitrogen ? [Re: peachy]
    #2697219 - 05/18/04 05:50 PM (12 years, 6 months ago)

It's really not that expensive.
But I guess it wont be as easy to store the mycelium in liquid nitrogen as it would be in a common fridge.
Would the mycelium not die when freezing since the cell walls must burst open when the water freezes in them?

Hmm, that would happen with your beer too, won't it? :frown:


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InvisibleSpeeker

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Re: Storing mycelium in liquid nitrogen ? [Re: ragadinks]
    #2697357 - 05/18/04 06:06 PM (12 years, 6 months ago)

You can use glycerin to prevent that happening at least in a normal freezer ..
Resurrecting a Better Method for Long-Term Storage of Mushroom Cultures

but the writer of that article prefers the idea of using distilled
water for storing cultures..


Edited by speeker (05/18/04 06:14 PM)


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Offlinepeachy
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Re: Storing mycelium in liquid nitrogen ? [Re: Speeker]
    #2697558 - 05/18/04 06:34 PM (12 years, 6 months ago)


Speeker:: Thanks for the link I'm going to give that a try. Distillation is simple enough :smile: And a chest freezer I already have.
Anyone else tried this? Might have to conduct some experiments to establish acceptable temperature norms for this sort of thing. It might also be a very viable way of transporting cultures.

ragadinks:: I suppose so, though I suppose you could just open them before putting them into your nitrogen freezer. Also I imagine you'd have to provide kevlar or equivalent gloves to your friends before passing them around; unless they're not that attached to the skin of their hands lol! K, no more puns.


--------------------
"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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OfflineNoG
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Re: Storing mycelium in liquid nitrogen ? [Re: peachy]
    #2698585 - 05/18/04 10:29 PM (12 years, 6 months ago)

I see this as being dangerous and unnessasary why store mycelium at this temp ? Its not like mushrooms are going to be extinct anytime soon. I just dont see the applications.


Later NoG


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Offlinepeachy
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Re: Storing mycelium in liquid nitrogen ? [Re: NoG]
    #2699404 - 05/19/04 01:54 AM (12 years, 6 months ago)

Unnecessary? Probably.
Dangerous? Certainly (without the correct environment for application of these techniques)

But still interesting, it'd be good for informational purposes to conduct some experiments and find out how much, for how long and at what cost psilocybin shrooms could be stored in such a fashion. If this is the way to statically preserve your favourite cultures for all time, I want to know about it :smile:


--------------------
"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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Re: Storing mycelium in liquid nitrogen ? [Re: ragadinks]
    #2701125 - 05/19/04 01:59 PM (12 years, 6 months ago)

Hi raga, check out the little thread posted lately 'freezing mycelia'... not sure if it is possible but I am heartened by your post that suggests that it is... at risk of repeating myself, the best bet would be to freeze them slowly (wrapped in a towel in the freezer, THEN into liquid nitrogen), to minimise formation of damaging water molecule aggregates, but thaw them fast, to maximise the breakdown of any of these that had formed... this is the strategy adopted by scientists freezing human lymphocytes or cancer cells etc... not all cell-type aggregates can be frozen and successfully brought back to life though (neural networks, for example, are pretty much rooted as soon as they're frozen, which is why 'cryogenic preservation' of peeps or even just their brains to be 'brought back later' seems such a scam to me!), and there is a chance it simply won't work with mycelia of some species... I would like some experienced shroom growers on here that have a 'spare cake' that is partly or newly colonised (and sterile) to simply freeze it (SOLID) for 2 or 3 days in a normal freezer, then thaw it out and nurture it, and see if they can bring it back to life... if they can, it suggests that liquid nitrogen storage could work well, at least for that particular species... maybe worth baring in mind though that temperature fluctuations (even below freezing temp) are bad for bio-sample viability, and if they actually thaw out then are re-frozen, more often than not the entire sample is cactus (not bacteria and viruses though)... which may present problems for the home grower trying to maintain liquid nit' type temps constantly, given the occasional malfunctions/ requirements for defrosting of some freezers etc... also, though you may well know this, peeps have reported in these pages that that they've accidentally frozen their spores-syringes solid in these pages, and most survived this process... spores are a lot hardier than mycelia though...
I can see definite advantages of being able to freeze mycelia... it would essentially mean that you could revisit shroom growing YEARS later, say, if your fave or only natural patches get wiped out/ have buildings buit on them/ desecrated in some way... also if you were into multiple strains, you could stash many away and be in no rush or need to 'maintain' them... it's also a lot quicker colonising a cake from myc than from spores, my guess is that this would even hold true if only 10% of the myc remained viable/ survived the freeze...
Again, I'm throwing up the challenge to someone that is experienced in growing (not me!) to freeze one of their cakes right through (just in a normal freezer), then see if they can bring it back to life... that would answer the question once and for all, re the ability of that particular species to withstand freezing...


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


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OfflineNoG
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Re: Storing mycelium in liquid nitrogen ? [Re: suboriginal]
    #2703571 - 05/20/04 12:26 AM (12 years, 6 months ago)

im guessing the strong would survive ? or the ones more resistant to being frozen. wouldnt this be isolating what your growing a bit like it could turn out to be only the crapy shrooms survived the freeze etc?


Later NoG


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Offlinepeachy
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Re: Storing mycelium in liquid nitrogen ? [Re: NoG]
    #2704119 - 05/20/04 02:30 AM (12 years, 6 months ago)

Well wouldn't hardier mycelia make for hardier shrooms? We could be pleasantly surprised.

I'm going to give this a try for the sake of experimentation, I've a 5litre cake (Ecuadorians) which'll be fully colonized in a few days that I've no real need for; so I'll bung it in the chestfreezer wrapped in a sterilized towel sealed in a ziplock bag and drop the temp as far as it'll go.


--------------------
"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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InvisibleSpeeker

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Re: Storing mycelium in liquid nitrogen ? [Re: peachy]
    #2704248 - 05/20/04 03:00 AM (12 years, 6 months ago)

So you're not going to mix it with glycerin first?
My guess is that it's gonna die without it...  :tongue:



Edited by speeker (05/20/04 03:57 AM)


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Offlineragadinks
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Re: Storing mycelium in liquid nitrogen ? [Re: peachy]
    #2704604 - 05/20/04 07:42 AM (12 years, 6 months ago)

At least it is worth a try.
Outdoor mycelium also freezes sometimes I guess.
But I think if you do it with liquid nitrogen you must add something like glycerin that keeps the cell walls from bursting.
It's a pity that we do not have any professional grower here that could tell us how it is done in the industry.


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InvisibleSpeeker

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Re: Storing mycelium in liquid nitrogen ? [Re: ragadinks]
    #2704633 - 05/20/04 08:07 AM (12 years, 6 months ago)



Edited by speeker (05/20/04 08:13 AM)


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Offlineragadinks
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Re: Storing mycelium in liquid nitrogen ? [Re: Speeker]
    #2704650 - 05/20/04 08:25 AM (12 years, 6 months ago)

Seems that it would be as simple as adding some glycerin to the culture - mix it up and deep freeze it in liquid nitrogen.
It's a pity that noone has made some experiences with liquid nitrogen in this forum.


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OfflineNoG
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Re: Storing mycelium in liquid nitrogen ? [Re: ragadinks]
    #2704762 - 05/20/04 10:03 AM (12 years, 6 months ago)

i dunno if id want to eat mushrooms off a Glycerin soaked cake. from what i understand you dont want to be eating glycerin. and since the stuff absorbs water its not really gonna be good for your 90% water mushies your trying to grow.

maybe soaking your cake in salt water to lower the freezing point of water would be a safer bet for a short freeze in the freezer. I wouldnt freeze the cake for long. and dunk in fresh water to rinse most of the salt off.


Later NoG


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InvisibleSpeeker

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Re: Storing mycelium in liquid nitrogen ? [Re: NoG]
    #2704886 - 05/20/04 11:07 AM (12 years, 6 months ago)

glycerin is used as sugar substitute as sweetner so you probably can eat it safely.. :wink:


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Offlinesuboriginal
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Re: Storing mycelium in liquid nitrogen ? [Re: Speeker]
    #2705848 - 05/20/04 03:25 PM (12 years, 6 months ago)

Sheesh, OK... glycerin would freeze at liquid nitrogen temps... it is used to store things without actually 'freezing them' at below zero, but not THAT far below zero!!! (like in a normal freezer, lets say... still minimises contams/ metabolism by contams ie. GROWTH of contams, but does not fracture the organelles of what you're trying to preserve, cause the solution doesn't actually freeze at 'moderately' below zero temps...
Second, storing your myc in glycerin (should someone on here try this) at below zero will not result in ingestion of glycerin when you eat the shrooms... the glycerin will be long gone 'in the chain; at that point...
Lastly, GOOD FREAKIN ONYA peachy... do the experiment, freeze that sucker, otherwise we're all just guessing... :-)
PS... freeze it slowly, thaw it quickly, then care for it carefully ... I'm on the edge of my seat for the results...

2nd lastly (!), re 'unnatural selection/ crappy shrooms' by freezing/ thawing/ re-nurturing, you could conceivably select for strains that might survive the cold winters in your own back yard, and re-emerge, say, a year after you had broken up a well colonised (frozen/ thawed/ nurtured) cake onto the right spot... just a wild guess...


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


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