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Offlinefossilshark
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The OFFICIAL Stasis Storage Thread * 25
    #28161262 - 01/28/23 12:06 PM (1 year, 4 months 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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OfflineVietboyX
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
    #28161273 - 01/28/23 12:12 PM (1 year, 4 months ago)

Nice write up, will definitely be helpful to link to the nubs in discord for long time storage :smile:

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Offlineside-eye
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Re: The OFFICIAL Stasis Storage Thread [Re: VietboyX]
    #28161281 - 01/28/23 12:16 PM (1 year, 4 months ago)

Great write up, been waiting for this!

What are you sterilizing the strips in after soaking, just any pp5 container?

Would using wood skewers still use the same process... soak to hydrate, pc, lay on a colonized plate, then into sterile distilled water?


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Offlinefossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: side-eye]
    #28161293 - 01/28/23 12:23 PM (1 year, 4 months 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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InvisibleNextcontestant
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
    #28161310 - 01/28/23 12:34 PM (1 year, 4 months ago)

:camping:

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Re: The OFFICIAL Stasis Storage Thread [Re: Nextcontestant] * 5
    #28161534 - 01/28/23 03:33 PM (1 year, 4 months ago)

Lids for drying out colonized grains for dry stasis

Ppl have also just left some kernels in the bottom of their qts (after securing the lid back on) and let them dry out that way but I hear it can take quite a while. 

I’m going to doing a lot of drying of cultures out in the future and probably simplify my lid set up so it’s just a small SHIP and use only two layers of tyvek with a widemouth 1/2 pint.. putting that in a bag of desiccant (damprid) with the larger lid and less barriers should speed things up by another week I’m thinking.  Right now I’m at about 3 weeks from hydrated to dry where I can store as is or load into swab packs to take up less space

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Re: The OFFICIAL Stasis Storage Thread [Re: fahtster]
    #28161548 - 01/28/23 03:45 PM (1 year, 4 months ago)

:popcorn:


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Offlinefossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: fahtster]
    #28161690 - 01/28/23 05:17 PM (1 year, 4 months 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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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark] * 2
    #28162481 - 01/29/23 08:01 AM (1 year, 4 months ago)

Interesting thread here. Gives me some ideas for when I’m gonna need to back up my library. Nice contribution :rockon:

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Invisiblebakedbeings
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Re: The OFFICIAL Stasis Storage Thread [Re: Pastywhyte]
    #28162493 - 01/29/23 08:13 AM (1 year, 4 months ago)

from what ive read so far it seems like i should be able to just take a clean tissue sample and stick it in some water, no?


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Re: The OFFICIAL Stasis Storage Thread [Re: bakedbeings] * 3
    #28162509 - 01/29/23 08:32 AM (1 year, 4 months ago)

Quote:

fossilshark said:
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?




Well there’s no visible growth after many months.. if there is cell division, it’s happening at a speed that isn’t noticeable after a certain point of dryness.  I just revived this mak 118 jar that’s looked exactly like this since the end of February




I think if you remove any part of those three, oxygen, food, water, you can get stasis or at least a degree of it.

That mak 118 is just about start knotting/pinning so we’ll see how it does.  You’d think after being dried out for 10 months it’d slow it down or wear out.  We shall see.  Ime, it does not


I actually already revived the clone once this season using a separate dried out jar but I was trying to laymix in a bag… didn’t work out so well and I had to remove the block from the container so fruit out the side pins.


So I kept it normal this time around as far as sub construction and I think it’ll be even better

But I also revived this fp+ culture that was dried for 8 months

Before..


After revival..

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Offlinefossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: bakedbeings]
    #28162739 - 01/29/23 11:36 AM (1 year, 4 months 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 (1 year, 4 months 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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Invisiblebakedbeings
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
    #28162757 - 01/29/23 11:44 AM (1 year, 4 months ago)

Quote:

fossilshark said:
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.



sweet :rockon:


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Invisibledna24
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
    #28162774 - 01/29/23 11:54 AM (1 year, 4 months ago)

i have been super interested in this since i first read about long term storage and currently am testing this myself. i am at nowhere the time frame to see if im doing it correctly yet but thats half the fun. i need to invest in some quality vials.


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Offlinefossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: dna24] * 1
    #28163355 - 01/29/23 06:45 PM (1 year, 4 months 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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Invisiblefungusul
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
    #28164960 - 01/30/23 08:02 PM (1 year, 4 months ago)

Thanks for sharing! :takingnotes:

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OfflineRange SpacerS
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
    #28166204 - 01/31/23 04:22 PM (1 year, 4 months ago)

Am I correct in my assumption that one cannot put liquid spores into stasis and that it would be better just to keep a spore print?


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Edited by Range Spacer (01/31/23 04:24 PM)

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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
    #28166219 - 01/31/23 04:37 PM (1 year, 4 months ago)

:headbang3:


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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 (1 year, 4 months 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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