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Offlinefossilshark
DouchebagDonny
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The OFFICIAL Stasis Storage Thread * 21
    #28161262 - 01/28/23 12:06 PM (11 months, 23 days ago)

I've been waiting a long time to make this thread as this is something I am very interested in. The goal of this thread is to share the many different methods of storing cultures in stasis. I will be posting my method and I would love for everyone to contribute their method of storing cultures in stasis. I in NO way came up with stasis storage and it has been around in many different forms for decades.

What is stasis storage?
Stasis storage is storing a sterile sample of mycelium in a no-oxygen no-nutrient environment causing the mycelium to go into stasis ceasing all metabolic activity.

Why stasis storage?
There are multiple advantages of stasis storage over other long term culture storage methods.

#1: The biggest advantage of stasis storage is the complete inaction of metabolic activity. Other long term culture storage methods (slants, cryo) only slow the metabolic activity down. In theory this can lead to changes in the culture that is not desired. Furthermore this means that unlike slants you do not have to "wake up" the mycelium at any point to continue to preserve it. Stasis has been shown to be effective for 30 years in https://sci-hub.st/10.1139/w08-049 and thats just when they stopped the study. In theory stasis storage is indefinite.

#2: Stasis storage is super simple and can be done with solid media or liquid or just straight up agar wedges. Very universal technique for anyone's style.

#3: My favorite part, stasis storage works at room temperature. Absolutely no need to take up valuable space in your moms fridge. :wink:

My method:

My initial trials 2 years ago were done to preserve Enigma x Rusty Whyte F2 from Solipsis using paper strips. I plan on switching over to wood skewers to make things easier to handle. First I hydrated paper strips by dunking paper strips in distilled water for a few seconds and sterilizing them for 20 minutes in my trusty presto along with some vials of distilled water.

After the paper strips and water were prepped I laid the strips on colonized germ plates (more strains more better baby) and let the mycelium colonize them for about a week.



After the paper strips were colonized I transferred them into vials of distilled water (along with some fat plate pins to try and clone them in the future).



And done. Thats the whole tek. Stasis storage is stupid simple. I ran the cultures about a year after I made them and they bounced back very quickly and I plan on doing a 2 year test next month when I have time.

NOTES: While the original stasis vials I made are useable its apparent that my sterile technique back then was sub par. One of the vials had a scoby on top but the mycelium was just fine.

For coprophilic species paper strips, wood skewers, toothpicks ECT work just fine with no issues. I theorize that if you want to store lignophilic species this would be too much available nutrients and effectively make the solution into LC. This would still work as once LC runs out of nutrients it goes into stasis, but to me that does not seem ideal. A better solution would be to use SFD discs like in the original paper.

I appreciate any questions or discourse on this thread on the topic. I think this method is an amazing tool for a mycology enthusiast. I appreciate those of you who take the time to read through this post. :mushroom2:


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Offlinefossilshark
DouchebagDonny
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Re: The OFFICIAL Stasis Storage Thread [Re: side-eye]
    #28161293 - 01/28/23 12:23 PM (11 months, 23 days ago)

I use pyrex test tubes from kartersci. Yes skewers would work exactly the same and I will be switching over to those for future cultures since they are easier to handle then paper strips.


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Offlinefossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: fahtster]
    #28161690 - 01/28/23 05:17 PM (11 months, 23 days ago)

This method is really interesting, so you think the mycelium goes into stasis despite the presence of oxygen? So basically it isn't exclusively no-oxygen no-nutrient but can also just be caused by a lack of water?


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Offlinefossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: bakedbeings]
    #28162739 - 01/29/23 11:36 AM (11 months, 22 days ago)

Baked that is correct. If you get a sterile sample of tissue you can just put it in sterile water and boom that clone is saved for generations to come.


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Offlinefossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: fahtster]
    #28162740 - 01/29/23 11:37 AM (11 months, 22 days ago)

Wow super dope. I might save some cultures like this in the future. I am very excited to run those gandalf grains you sent me a while back.


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Re: The OFFICIAL Stasis Storage Thread [Re: dna24] * 1
    #28163355 - 01/29/23 06:45 PM (11 months, 22 days ago)

Thats why this thread is 2 years in the making. I wanted to make sure it works before I started posting about it.


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Offlinefossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: Range Spacer]
    #28167021 - 02/01/23 03:31 AM (11 months, 19 days ago)

Technically speaking (I may be wrong about this) because spores by themselves with no nutrient will not reproduce they are already in stasis.

As far as I know spore syringes probably wouldn't stay viable for infinitely long periods of time even with proper storage, implying that it is either not true stasis or there is other factors that prevent spores from germinating after X amount of time.

Why not just germinate those spores and put the resulting culture into stasis?


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Offlinefossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: LadysKnight]
    #28167217 - 02/01/23 08:24 AM (11 months, 19 days ago)

Probably lol. I definitely didnt lose the genetics tho as ive been able to revive them on 2 seperate occasions.


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Offlinefossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: towndaze]
    #28169710 - 02/02/23 07:30 PM (11 months, 18 days ago)

Not a clue what you just said but hell yes brother I love the energy.


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Re: The OFFICIAL Stasis Storage Thread [Re: nektar61] * 3
    #28174161 - 02/05/23 09:53 PM (11 months, 15 days ago)

Spores are just gametes and not fully developed cells so they wouldnt have the resources to completely shut down and stay viable.

Based on what methods people use at the moment if you get rid of just one of the three oxygen, water, or nutrients the mycelium will go into stasis and be viable until it is either destroyed or wakes up.

Stasis storage of mycelium cultures needs no overcomplicating.


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Re: The OFFICIAL Stasis Storage Thread [Re: Rushing] * 1
    #28355329 - 06/11/23 08:30 AM (7 months, 13 days ago)

Love seeing everyone's take on stasis storage! Thats exactly why I created this thread, would love to see more!


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Re: The OFFICIAL Stasis Storage Thread [Re: DredgeMyEyes] * 1
    #28358195 - 06/13/23 10:01 AM (7 months, 11 days ago)

The medium you use to get myc into your stasis vials makes no difference in the long run. Choose something that you feel comfortable handling.

As far as too much nuts on a medium like an agar wedge, even if the vial does become LC at some point it will run out of nuts and go into stasis. The only concern with this is possibility of it becoming too thick to get out easily but i doubt that would be a concern with just a slice of agar.


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Re: The OFFICIAL Stasis Storage Thread [Re: Screwup] * 1
    #28360775 - 06/15/23 12:15 PM (7 months, 9 days ago)

I use bottled water just because its what I have on hand.

As for wooden pieces I am finding that putting boiled and sterilized pieces of wood skewers does work but takes a long time for myc to colonize it. I do not know how well it would work just putting a wedge or a bit of LC on it.


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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark] * 3
    #28405356 - 07/23/23 03:55 PM (6 months, 2 days ago)

Threw some Gymnopilus Purperatus and Gandalf in stasis today, decided to switch over to wood sticks as they are way easier to handle then paper.

I wonder how necessary the water even is?





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