Home | Community | Message Board
You are not signed in.
Sign In New Account
Forum Index Search Posts Trusted Vendors Highlights Galleries FAQ User List Chat Store Random Growery

Magic-Mushrooms-Shop.com
This site includes paid links. Please support our sponsors.

Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
Threaded Mode Threaded  Previous topic Previous  View all topics Index  Next topic Next 

Welcome to the Shroomery Message Board! You are experiencing a small sample of what the site has to offer. Please login or register to post messages and view our exclusive members-only content. You'll gain access to additional forums, file attachments, board customizations, encrypted private messages, and much more!

Shop: Myyco.com Isolated Cubensis Liquid Culture For Sale   MagicBag.co All-In-One Bags That Don't Suck   Mushroom-Hut Liquid Cultures   Original Sensible Seeds Bulk Cannabis Seeds   North Spore Injection Grain Bag   Sporeworks.EU Spores for European Microscopy

Jump to first unread post Pages: | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 |  [ show all ]
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: ReverendMyc] * 1
    #28910261 -

Probably would be good for just  shaking the vial and pour it onto a couple brf pucks too


--------------------

Links For Beginners And Beyond:mspoil:

Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: rumfor69] * 1
    #28918618 -

Thanks for the write-up Mycoplex! super simple method, I would much rather do this than make slants

I have stored a bunch of cultures this way over the last couple years. I've had trouble with reviving a lot though. Some months ago I think I tried reviving like 6 and 4/6 were just real slow growing and then eventually died out.

Has anyone else had a better success rate with older cultures?
I've been using autoclaved milli-Q water, maybe that's causing issues by being too pure :shrug:

Planning to revive a whole bunch of them pretty soon to see how many grow. I'll post the results here


--------------------

Edited by TheDoobsker (08/21/24 12:24 PM)

Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: TheDoobsker] * 1
    #28918701 -

Just did 3 different varieties from a clean t0, 3 small cubes of agar in 5ml tubes. I figured the more water the better when it comes to putting them into stasis. At least that's what I'm telling myself 😅


--------------------
15 year lurker

Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: TheDoobsker]
    #28919906 -

Great work fellas, I also have a few updates which I'll write here as an extension of what Doobsker is asking above (success with old cultures, etc.).

TheDoobsker said:
Has anyone else had a better success rate with older cultures?



A couple weeks ago I restored a cryovial culture from early 2022 of Tasmanian cubensis and since then I was able to expand it to more agar, LC, grain, and more cryovial backups in a matter of one week.  The culture grows on agar, grain, and LC just the same as it would if I had isolated it recently, it's no slower and is no less vigorous.  Actually the speed at which it revived and expanded was how I was able to recreate the entire culture from a stored vial.

I started with the restoration of the vial a few agar cups like I wrote a few replies back.  After about 4 days, I used that same growth (no need to isolate it further because the culture is already isolated, you're just reviving it) to expand to six more agar plates, two jars of LC, and  several grain jars.  I also recreated the backup by making 5-6 more cryovials.  All of the expansions are growing vigorously and quickly.

So with a single resurrection I expanded the stored Tasmanian culture out and also saved it again to more cryovials, and in the entire time that the cryovial was stored over the last two years it used no electricity or special storage, it was just in a vacuum sealed bag at room temp.

If you're experiencing a culture that stalls out or dies after the resurrection, maybe try a different agar recipe, or try putting it directly to an LC, etc.


--------------------
Cryovial Tek
Indoor Ovoid Community
ATL7 Cosmoplex Gallery
Fungus Gnat Control Tek
Precise Pasteurization Tek

Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Mycoplex] * 1
    #28919918 -

Odd question but how was it stored inside the actual vial, one big slab of agar like you have pictured or did you use a different method?


--------------------
15 year lurker

Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Meanwhile]
    #28920220 -

Meanwhile said:
Odd question but how was it stored inside the actual vial, one big slab of agar like you have pictured or did you use a different method?


The amount of agar I place into each vial is about the same as the small piece shown in the front page in the photo with the scalpel.  The piece I put in is quite small and sometimes I have to look twice in the vial to see it in the first place.


--------------------
Cryovial Tek
Indoor Ovoid Community
ATL7 Cosmoplex Gallery
Fungus Gnat Control Tek
Precise Pasteurization Tek

Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Mycoplex] * 3
    #28956462 -

Update:  I've successfully fruited the cubensis culture above that was resurrected from 2 years ago (put into stasis early 2022).  My experience with this is no different than if the culture was newer or recently isolated, all the timetables were the same and the fact that this was an aggressive colonizer still came through even though it was in stasis for 2 years.



--------------------
Cryovial Tek
Indoor Ovoid Community
ATL7 Cosmoplex Gallery
Fungus Gnat Control Tek
Precise Pasteurization Tek

Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Mycoplex] * 1
    #28956523 -

Mycoplex said:
Update:  I've successfully fruited the cubensis culture above that was resurrected from 2 years ago (put into stasis early 2022).  My experience with this is no different than if the culture was newer or recently isolated, all the timetables were the same and the fact that this was an aggressive colonizer still came through even though it was in stasis for 2 years.




That is awesome.

Was there any extra time for initial growth on the plate from cold storage?

Nice to know the culture wasn't affected.

Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Bigdogg] * 1
    #28956540 -

Bigdogg said:
Mycoplex said:
Update:  I've successfully fruited the cubensis culture above that was resurrected from 2 years ago (put into stasis early 2022).  My experience with this is no different than if the culture was newer or recently isolated, all the timetables were the same and the fact that this was an aggressive colonizer still came through even though it was in stasis for 2 years.




That is awesome.

Was there any extra time for initial growth on the plate from cold storage?

Nice to know the culture wasn't affected.


Yes, there was a period of about 3 days where I was waiting for it to "wake up" after popping it out of the vial.  Once it kicked off the standard timetables applied.  Thanks for checking out this thread!


--------------------
Cryovial Tek
Indoor Ovoid Community
ATL7 Cosmoplex Gallery
Fungus Gnat Control Tek
Precise Pasteurization Tek

Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Mycoplex] * 1
    #28956583 -

Thanks for making this thread.

Coming back from a long break away from the hobby I wish I had this info back then. Trying to bring back 5+ year old spores has been frustrating.

Good work with the write-up, testing and long term commitment to the project. 👍👌

:cool:

Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Mycoplex] * 1
    #28956639 -

Mycoplex said:
Update:  I've successfully fruited the cubensis culture above that was resurrected from 2 years ago (put into stasis early 2022).  My experience with this is no different than if the culture was newer or recently isolated, all the timetables were the same and the fact that this was an aggressive colonizer still came through even though it was in stasis for 2 years.




Nice homie :thumbup: really appreciate the updates


--------------------

When we constantly pull things apart trying to see how it works, we may end up with only an understanding of how to destroy something
- nick sand


Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Psicomb] * 1
    #28962128 -

What was the longest revival time you guys had to wake them up?

Ive put some 2-3months old wedges from vials to water agar and low nutes agar and non had taken off after 7 days so far.

I started experimenting with this method 2 years ago or so, but after a year or two, in that case i used scraped myc, most of cultures died. Back then i was storing cubensis and myc turned kinda brownish. I used store bought destilled water which is actually demineralised water so i thought that is the reason. So this year i made my own destilled water, actually destilled and stored some pans.

The myc on agar wedges in vials looks nice and white after 2-3 months.
But no growth after 7 days so far with these.
Only one that woke up was copelandia i had stored for a year, only myc in water in vials. That one is still with store bought demineralised water.

Previous experiments with scraped myc with pans didnt work out well, myc got all bruised and black after few weeks. Except for one sample, copelandia mentioned above.


The rest are ttbvi's and weza's and as i said are from June and July this year.
Maybe too little nutes? My low nutes agar is 0.5% lme only.

Should they take off on water/low nutes agar? Well copelandia did in 3-4 days or so..

Edited by Rush56 (09/26/24 08:33 AM)

Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Rush56] * 1
    #28962562 -

Rush56 said:
What was the longest revival time you guys had to wake them up?


In this thread I recently resurrected a 2 year old culture and fruited it (above).  I am sure that the timeframe can be significantly longer than that, and I'm sure that throughout the years others have done this with older samples.

My goal was to show it real-time in this thread, so I revived the same samples I was describing in the writeup on the first page.

I'm not sure about reviving them on water or low nutrients, if you're confident about that approach keep tinkering with it.  I drop the samples directly onto potato flake agar and they revive in about 4 days


--------------------
Cryovial Tek
Indoor Ovoid Community
ATL7 Cosmoplex Gallery
Fungus Gnat Control Tek
Precise Pasteurization Tek

Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Mycoplex] * 1
    #28963167 -

I will be reviving some 2 year old cryo vials in the next couple months; will update then.  Expecting it to go just fine :thumbup:


--------------------

When we constantly pull things apart trying to see how it works, we may end up with only an understanding of how to destroy something
- nick sand


Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Psicomb] * 1
    #28963432 -

Psicomb said:
I will be reviving some 2 year old cryo vials in the next couple months; will update then.  Expecting it to go just fine :thumbup:


Psicomb, nice to see ya man.

I am dumb so pardon this question. What's the best way to get the wedge out of the vial? Sterile swab?


--------------------
Cultivating Panaeolus cyanescens


Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Asura]
    #28964358 -

Asura said:
Psicomb said:
I will be reviving some 2 year old cryo vials in the next couple months; will update then.  Expecting it to go just fine :thumbup:


Psicomb, nice to see ya man.

I am dumb so pardon this question. What's the best way to get the wedge out of the vial? Sterile swab?


Yo, Asura!  There are multiple techniques that all should work, here are a few, if you have more ideas  we can also list those.
  • Use an inoculation loop to scoop out some samples.  You can shake the vial before doing this to break up the myc further.
  • Use a sterilized scalpel to scoop out a wedge
  • Shake the vial and then gently pour some into an agar plate without using any tools.  This is the technique I last used on the 2 year old sample and I found that all it took as a few drops from the vial to get the sample to revive.  The wedge itself doesn't need to come out, just some of the water in the vial.
  • Use a sterilized syringe + needle to suck some of the liquid out after agitating the vial.


--------------------
Cryovial Tek
Indoor Ovoid Community
ATL7 Cosmoplex Gallery
Fungus Gnat Control Tek
Precise Pasteurization Tek

Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Mycoplex] * 1
    #28964369 -

Thanks for for this write up. Picked up some vials the other day, did a small batch yesterday. 3D printed a little vial holder for them.




Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Henry Swanson] * 1
    #28964388 -

Henry Swanson said:
Thanks for for this write up. Picked up some vials the other day, did a small batch yesterday. 3D printed a little vial holder for them.






Epic!  Thanks for sharing this in the thread.  That's a really useful project and others might get the idea to replicate.


--------------------
Cryovial Tek
Indoor Ovoid Community
ATL7 Cosmoplex Gallery
Fungus Gnat Control Tek
Precise Pasteurization Tek

Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Asura] * 2
    #28965029 -

Asura said:
Psicomb said:
I will be reviving some 2 year old cryo vials in the next couple months; will update then.  Expecting it to go just fine :thumbup:


Psicomb, nice to see ya man.

I am dumb so pardon this question. What's the best way to get the wedge out of the vial? Sterile swab?


I wipe the outside of the vial down with iso, very gently pour out the water into a sterile dish but leave the culture behind in the vial and then just tap the vial upside down til wedge(s) falls out in a plate.  It isn’t perfect but surprisingly I never get contams this way. 

I’m just lazy and half-caveman, so that’s my approach :shrug: guarantee there’s better ways out there lol

Oh and hey, Asura! so good to still see you posting :sun:


--------------------

When we constantly pull things apart trying to see how it works, we may end up with only an understanding of how to destroy something
- nick sand


Extras: Filter Print Post Top
Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Psicomb] * 1
    #28965081 -

Mycoplex said:
Yo, Asura!  There are multiple techniques that all should work, here are a few, if you have more ideas  we can also list those.
  • Use an inoculation loop to scoop out some samples.  You can shake the vial before doing this to break up the myc further.
  • Use a sterilized scalpel to scoop out a wedge
  • Shake the vial and then gently pour some into an agar plate without using any tools.  This is the technique I last used on the 2 year old sample and I found that all it took as a few drops from the vial to get the sample to revive.  The wedge itself doesn't need to come out, just some of the water in the vial.
  • Use a sterilized syringe + needle to suck some of the liquid out after agitating the vial.



Psicomb said:
I wipe the outside of the vial down with iso, very gently pour out the water into a sterile dish but leave the culture behind in the vial and then just tap the vial upside down til wedge(s) falls out in a plate.  It isn’t perfect but surprisingly I never get contams this way. 

I’m just lazy and half-caveman, so that’s my approach :shrug: guarantee there’s better ways out there lol

Oh and hey, Asura! so good to still see you posting :sun:


These are great options so thanks guys. It took me a minute to figure out a good way
to get the wedges in...now for the other direction. This type of work is hard when
the eyesight isn't so good anymore.


--------------------
Cultivating Panaeolus cyanescens


Extras: Filter Print Post Top
Jump to top Pages: | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 |  [ show all ]

Shop: Myyco.com Isolated Cubensis Liquid Culture For Sale   MagicBag.co All-In-One Bags That Don't Suck   Mushroom-Hut Liquid Cultures   Original Sensible Seeds Bulk Cannabis Seeds   North Spore Injection Grain Bag   Sporeworks.EU Spores for European Microscopy

Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
Threaded Mode Threaded  Previous topic Previous  View all topics Index  Next topic Next 

Similar ThreadsPosterViewsRepliesLast post
* Best short-term storage... TheShroomHermit 2,115 11 05/03/03 04:32 PM
by Prisoner#1
* Best printing tek for storage? TheDoc 1,356 8 09/23/03 04:36 PM
by shirley knott
* short term storage honeyroasted 1,507 2 10/01/01 11:42 PM
by MushroomMatt
* Re: Honey Water Tek Anonymous 795 4 04/19/00 06:40 PM
by Hippie3
* Shot Down By An Expert! *DELETED* *DELETED* *DELETED* Amanita_Dreamer 1,039 10 05/16/03 11:28 PM
by Ali
* -=Poor Mans Syringe Tek=-
( 1 2 3 4 all )		 LiL_KuSsH 8,993 74 05/02/04 04:43 PM
by Rose
* Storage of mushrooms in vacuum-sealed bag... Horseman 4,262 5 07/25/03 08:32 PM
by govna
* Drying TEK JohnnyRespect 677 2 04/19/03 07:51 AM
by Prisoner#1

Extra information
You cannot start new topics / You cannot reply to topics
HTML is disabled / BBCode is enabled
Moderator: DERRAYLD, Shroomism, george castanza, RogerRabbit, veggie, mushboy, fahtster, the_chosen_one, LogicaL Chaos, 13shrooms, hamloaf, cronicr, Stipe-n Cap, Pastywhyte, bodhisatta, Tormato, Land Trout, A.k.a
13,869 topic views. 10 members, 2,022 guests and 202 web crawlers are browsing this forum.
[ Show Images Only | Sort by Score | Print Topic ]
Search this thread:

Copyright 1997-2026 Mind Media. Some rights reserved.

Generated in 0.039 seconds spending 0.008 seconds on 16 queries.