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fossilshark
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The OFFICIAL Stasis Storage Thread 21
#28161262 - 01/28/23 12:06 PM (11 months, 23 days ago) |
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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. 
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.
-------------------- LITFA LITFA LITFA
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fahtster
Now With 33%More Faht



Registered: 06/17/06
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Re: The OFFICIAL Stasis Storage Thread [Re: Nextcontestant] 4
#28161534 - 01/28/23 03:33 PM (11 months, 23 days ago) |
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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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side-eye
potate


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Re: The OFFICIAL Stasis Storage Thread [Re: bakedbeings] 4
#28181951 - 02/11/23 08:50 AM (11 months, 9 days ago) |
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I dig it so I tried it.
I got some 5mL tubes with a rack, and wood stir sticks. Each one breaks into three or four pieces that fit in the tubes. Then I filled a bunch of the bottles with distilled water.


I soaked the stir sticks in the bottle for about 30 minutes, figured they needed a little more time than paper. Poured water out of the stir stick bottles, then PC'd everything in my IP for 45 minutes. Stir sticks onto a few plates. These pictures were taken two days apart, myc is already starting to get into the wood. Be sure all the broken pieces are ripped apart from each other, you can see how the Reishi plate has a big V shape that I didn't want to risk pulling apart.
 
Once this grows into the wood I'll drop em into the distilled water. I'll report back in 30 years to check if your tek actually works.
-------------------- LAGM 2023
Edited by side-eye (02/11/23 08:51 AM)
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fahtster
Now With 33%More Faht



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Re: The OFFICIAL Stasis Storage Thread [Re: fahtster] 4
#28182962 - 02/11/23 07:45 PM (11 months, 9 days ago) |
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Quote:
fahtster said: 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

Bag did aight.

The question is if it’s able to go 10 months dried out at room temp and come back and do that ^^^ what’s stopping it from going 10 more, or even more than that?
I have dried out cultures in jars that are over a year old. I can revisit this thread in a couple years and give those cultures a try at reviving.. see ya then
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Guerrilla
Bumbaclart


Registered: 01/30/21
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Re: The OFFICIAL Stasis Storage Thread [Re: side-eye] 4
#28190466 - 02/16/23 04:17 PM (11 months, 4 days ago) |
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What's the purpose of using wood or paper when a wedge of myc will suffice and last indefinitely? Seems like you're making it way more complicated than it needs to be.
-------------------- Being pissed on does not make you a real man.
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the_chosen_one
On the Darkslide


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Re: The OFFICIAL Stasis Storage Thread [Re: 3.A.M] 4
#28609668 - 01/04/24 04:40 PM (23 days, 8 hours ago) |
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How is it a person can be here for years and all the sudden something like this pops up and it was started a year ago? 
I've become a huge fan of the 2ml cryovials and the Castellani method.

OR
Just forget some syringes in the fridge for 13 years. We'll see these in a decade or so.

Works for spores though. I've labelled these Tamp 13 for just that reason lol.

Thanks to Tree Frog for the link!
-------------------- "Luck favors the observant." - Workman
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fossilshark
DouchebagDonny



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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) |
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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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nektar61
Into SporePlay



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Re: The OFFICIAL Stasis Storage Thread [Re: side-eye] 3
#28182942 - 02/11/23 07:32 PM (11 months, 9 days ago) |
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Quote:
side-eye said: I'll report back in 30 years to check if your tek actually works.
Excellent.
I have an experiment I've set up on here that should last 15 years. (vacuum packing very dry clone shrooms into 2 gram packs, then taking one every year and reporting back. First one will be pretty soon.
-------------------- -NEW? Start here.
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fossilshark
DouchebagDonny



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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) |
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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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astra
Mycelia-Of-House-Fungi



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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark] 3
#28410278 - 07/27/23 11:43 AM (5 months, 29 days ago) |
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dH₂O 2ml cryovials

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fahtster
Now With 33%More Faht



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Re: The OFFICIAL Stasis Storage Thread [Re: Screwup] 3
#28486326 - 09/29/23 12:01 AM (3 months, 28 days ago) |
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Getting to be about that time of year for me. Started reviving some dried out grain cultures 8 days ago. I GLC the dry grains and inject them into fresh grains. I can (and sometimes do) go straight to qts but I like to test the cultures first in small jars just to be sure.
Claketi (dried out April 2023)


PE x RW f7 clone (dried out March 2023)


Mak TP (this one is from March 2021)


Fp+ f8 clone (dried out April 2023)


Shit amazes me every time. These new jars will be used to glc to qts for tubs. When I made these new jars, I made a second one of each that will be dried out again for further storage
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Nichrome
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Re: The OFFICIAL Stasis Storage Thread [Re: bongoman] 3
#28509851 - 10/18/23 06:47 PM (3 months, 8 days ago) |
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I store lc in sealed vials for years on end. It's not exactly the same as a wedge in water but is similar conceptually. I have a Cordyceps that still kicks off like it should from just one drop from the banked vial. Same with semperviva, tamp, etc. I'd make a solid edumacated guess that a nutritious agar wedge in water would work just fine and there is no need for special agar prep for such a storage method.
I've cut pieces of stem and kept them in baggies for 6 months before getting around to plating them as clones. Mycelium, circumstantially, is capable of extreme longevity.
-------------------- “Better to be deprived of food for three days, than tea for one.”
Freedom is not the right to do as you please, but the liberty to do as you should. ~Emerson
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Pastywhyte
Say hello to my little friend



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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark] 2
#28162481 - 01/29/23 08:01 AM (11 months, 22 days ago) |
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Interesting thread here. Gives me some ideas for when I’m gonna need to back up my library. Nice contribution
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fahtster
Now With 33%More Faht



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Re: The OFFICIAL Stasis Storage Thread [Re: bakedbeings] 2
#28162509 - 01/29/23 08:32 AM (11 months, 22 days ago) |
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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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Bigworm



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Re: The OFFICIAL Stasis Storage Thread [Re: bongoman] 2
#28426269 - 08/08/23 08:22 PM (5 months, 17 days ago) |
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Cryo storage works well. I can't say I would trust anything but distilled water with my cultures but I'm sure it's worth a try. I also made mini slants with boiled toothpicks and agar for a shorter storage method compared to the distilled cryo storage.
Edited by Bigworm (08/08/23 08:22 PM)
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fossilshark
DouchebagDonny



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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) |
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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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Hindsight
Mad Scientist


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Interesting idea. I do the same thing but just take a piece of colonized agar and put it in sterile distilled water vial. It has worked well for me. There is, of course, a small amount of nutrient from the agar, but it doesn't seem to impact storage life. Paper is a nutrient source so it seems to be a roughly equal approach.
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bakedbeings
orbiter of truth


Registered: 09/01/20
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Quote:
Sulfurshelfsean said: I've had lcs last years. One out out clean mycellium for six years but eventually they contam. I always use very little nute so I always wondered if they were in stasis. After about a year the culture liquid is completely clear. This looks like a much better way to store thank you for the write up. I'm going to try this.
ive noticed the same thing with low nute LCs. they colonize about 80% and then freeze
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side-eye
potate


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Re: The OFFICIAL Stasis Storage Thread [Re: fahtster] 1
#28190459 - 02/16/23 04:09 PM (11 months, 4 days ago) |
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1
Alright so I got two plates' sticks in water.

I ran into a few problems on this one. First was I let the plates colonize the sticks a little too much, especially on the Reishi plate. I tried picking it up with tweezers, but it was so stuck to the plate that I lifted up the whole puck of agar out of the dish. I tried cutting it out but it still grabbed any agar underneath the stick. Reishi was by far the worst, but all of the sticks lifted a bunch of myc off the plate. Here's how colonized the plates were:

Second, the wood sticks float. These tubes are 5mL so pretty short, and when I let go of the stick it floated about halfway out of the tube. I used the lid to push them down as I capped em. Longer tubes/sticks may not be as bad, and paper may not be as bad either. I'm worried they picked up some bac hitting the rim of the tube, so I will give this another go soon with paper vs sticks.
Lastly, my fault, I dropped some sticks in the wrong tube of water. Not sterile so I had to toss those. Don't work high, kids.
-------------------- LAGM 2023
Edited by side-eye (02/16/23 04:10 PM)
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dna24
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark] 1
#28254230 - 03/30/23 07:53 AM (9 months, 24 days ago) |
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Quote:
fossilshark said: 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.
THIS, this is the way.
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 FUCK STIPE 
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