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How do YOU store your samples? OR Interesting Storage Examples
    #16714404 -

Sorry if this should be under another section, but I though advanced mycology fit it best, as it's essentially a secondary "skill" to the usual mushroom cultivation.

I'm wondering how people here store their syringes, LC, prints, petri's, etc.  Both long or short term if they are somewhat interesting or useful. I'm not asking about just normal storage conditions- I've read many different guidelines about temp/light/etc. found in the forums and the mushroom info section; I'm more looking for things like ways people might have rigged a humidor to hold syringes instead of cigars. Some might say I'm looking for weird or strange storage examples, while others would possibly say ingenious or the like for some of the better examples.

Personally I'd love to find some antique medical/science equipment that might store some things, though price and selection make this unlikely, at least for now. I've not seen anything specific yet, but I'm looking some now that the idea came to mind. I thought about that after remembering some of the old needle kits and whatnot from 60-120 years ago. Looking through old medical stuff I've seen many interesting specimen containers, and lots of stuff I don't even know the name of, but I thought might look cool re-purposed. I would also love to do a steam-punk style case of sorts at some point :tongue:

I'm likely a ways away from storing anything of mine in one of these ways, but the longer I plan/think about it the better my options might be. Another idea I'd love to have the time/equipment/money to do: I could rig a small suitcase or something to automatically regulate, and control the samples; that is I'd place them into a slot, it pulls them through, scans the label, saves the label and marks the sample storage slot for later. A little led screen with two or three buttons could be used to make my selections, and once made the samples can rotate around till it finds the right one and out it pops. I know there are plenty of modern day machines that'd do this, but I don't have the funds to buy one, and I find it much more enjoyable to build something like that myself. I'd want to find a good way to make it maintain the right temps and humidity too.


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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: otakun]
    #16715039 -

i store them in half pint mason jars

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: florida_guy]
    #16719211 -

After experimentation with various long and short-term techniques, I now use "industry standard" methods. Long term storage is done with slants, short term with agar plates. I toss my agar plates when they reach 6 months of age, and re-culture my slants at 18 months. Both are stored in a refrigerator set to about 34 degrees F. Tropicals are stored at higher refrigerator temperatures. If you don't refrigerate cultures, they won't last.

Automatic selection of samples??? I keep my 160+ strains (2 samples each = 320+ tubes) in plain old test tube racks, sorted in alphabetical order by Latin name. I used to use computer-printed labels, but found it to be too much trouble. Now I just mark the slant tubes with a felt-tipped pen.


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Liberté, égalité, humidité.

Edited by Terry M (08/18/12 06:54 AM)

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: otakun]
    #16726324 -

Hi Otakun,

I have used the following methods:

http://www.mycosphere.org/pdfs/MC1_3_No1.pdf

http://www.cssm.info/priloha/fm2008_534.pdf

https://files.shroomery.org/cms/burds94a.pdf


Regarding your idea of an automated machine, I've never seen anything like that before.

Cheers,


Wilson

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: wbastz]
    #16726741 -

wbastz said:
http://www.cssm.info/priloha/fm2008_534.pdf




This paper talks about using perlite as a long-term storage medium. It reports longer storage time than with slants. I have regular correspondence with the author, Ladislav Homolka. I'll ask Ladislav if he would absolutely trust his own CCBAS culture library with this method! :smile:


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Liberté, égalité, humidité.

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: Terry M]
    #16726822 -

Hi Terry,

I believe he uses liquid nitrogen and this method too !

Wilson

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: Terry M]
    #16726904 -

Hi Terry,

This link shows a pps file containing some photos on the use of perlite, but most of the text is written in Czech.

http://sun2.biomed.cas.cz/ccbas/CCBAS.pps

Apparently, he uses perlite as a support for the mycelium and stores these tubes in liquid nitrogen.

Cheers,

Wilson

Edited by wbastz (08/19/12 02:08 PM)

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: wbastz]
    #16727196 -

wbastz said:
Hi Terry,

This link shows a pps file containing some photos on the use of perlite, but most of the text is written in Czech.

http://sun2.biomed.cas.cz/ccbas/CCBAS.pps

Apparently, he uses perlite as a support for the mycelium and stores these tubes in liquid nitrogen.

Cheers,

Wilson


Thanks, Wilson!

Wow, I didn't know that cryogenic storage was so sensitive to culture medium. I assumed it was always 100%, except for maybe a few species. I have no experience with it, nor am I likely to unless the milkman starts delivering liquid nitrogen to me every week.
:lol:

Regards,
Terry


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Liberté, égalité, humidité.

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: Terry M]
    #16729086 -

The Czech file showing cryopreservation seems to predate the Long-term Maintenance file by about 7 years.  I want to try his method in any case. 

I am especially interested in his maintenance media formula of 4° Balling & 1.5% agar which equates to a 1.016 SG wort + 15g agar per liter.  My calculations show 41.4g of malt extract per liter which seems extremely high to me but I'll break out my brewing refractometer tomorrow (yes, I have been a home brewer for over seven years) and verify the amount of malt extract by weight per liter to equate to 1.016 SG (4°Balling).

I have been following Aloha's procedure of using a coffee stir stick (made from hardwood) in every slant.  John's theory is that after the agar media is exhausted the mycelia can live off of the wood for an extended period of time. 

It will be interesting to test the two methods side-by-side using the same agar media.


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"Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin

"Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: OICU812]
    #16729873 -

The wooden coffee stirrer or piece of tongue depressor for slants is pretty standard around here. RR teaches it in his video.


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Liberté, égalité, humidité.

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: Terry M]
    #16730056 -

I've also seen agar formulas containing salt.  As a matter of fact I used one I found (and modified) recently.  The mycelium seem to be accepting it better than I thought. 

I have read others use of freezing with success.  It would be good to hear from them too . . . hint hint.  I found this interesting article by James Harris, PhD that I want to try too.  In his article, Dr. Harris claims " This technique is highly successful. Many fungi (including filamentous molds, yeasts and yeast-like fungi) will survive for a decade or more in sterile water."


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"Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin

"Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson

Edited by OICU812 (08/20/12 07:05 AM)

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: OICU812]
    #16730247 -

Quote:
OICU812 said:

I am especially interested in his maintenance media formula of 4° Balling & 1.5% agar which equates to a 1.016 SG wort + 15g agar per liter.  My calculations show 41.4g of malt extract per liter which seems extremely high to me but I'll break out my brewing refractometer tomorrow (yes, I have been a home brewer for over seven years) and verify the amount of malt extract by weight per liter to equate to 1.016 SG (4°Balling).




According to my ProMash brewing software, 1 liter of water will require 50.92g of Malt Extract to formulate a wort with a specific gravity of 1.016.  This still seems too high to me.  I will verify this with a refractometer when I create more media lather this week.

Terry, if you would, when you correspond with Ladislav Homolka, please ask him if this concentration is still in use by them.


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"Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin

"Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: OICU812]
    #16731288 -

Since I don't have a cryogenic chamber, I've been doing two versions of each strain- 1 slant on agar w/ toothpick (a la RR), 1 wedge in water.  Never heard of salt, but from what I've read these two methods seem to work better than pure agar for long-term.

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: nycomyco]
    #16761713 -

In doing some searches today, I ran across this article (I don't think it's a post) that seems to have been written in  April 1998 by: Joseph C. Kish and posted/uploaded in the shroomery 02/17/03.  This article confirms the sterile distilled water method with some additional data.

Resurrecting a Better Method for Long-Term Storage of Mushroom Cultures

I also found this list of articles:

Agar and Culture Storage


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"Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin

"Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson

Edited by OICU812 (08/30/12 06:50 PM)

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: OICU812]
    #16762150 -

OICU812 said:
In doing some searches today, I ran across this article (I don't think it's a post) that seems to have been written in  April 1998 by: Joseph C. Kish and posted/uploaded in the shroomery 02/17/03.  This article confirms the sterile distilled water method with some additional data.

Resurrecting a Better Method for Long-Term Storage of Mushroom Cultures

I also found this list of articles:

Agar and Culture Storage


I've read the Kish paper, and others. I'd be happy to send you those, as they included very specific procedures. I tried sterile DW is a few different forms, over more than a year's time. At first, I thought it would save me a ton of work, since SDW appeared to have a longer storage life than slants. And I've got more than 150 different edibles strains to maintain.  But it's just not 100% reliable, like slants are. And you need 100%, or you risk losing a culture. So better make slants for backup! :smile:


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Liberté, égalité, humidité.

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: Terry M]
    #16762508 -

Quote:
Terry M said:

I've read the Kish paper, and others. I'd be happy to send you those, as they included very specific procedures. I tried sterile DW is a few different forms, over more than a year's time. At first, I thought it would save me a ton of work, since SDW appeared to have a longer storage life than slants. And I've got more than 150 different edibles strains to maintain.  But it's just not 100% reliable, like slants are. And you need 100%, or you risk losing a culture. So better make slants for backup! :smile:


I wholeheartedly agree, Terry.  Redundancy is in my blood.  I have had quite a few system failures, mostly PC (computer) related, and I have learned the hard way that multiple backups are essential.

I also agree that this method would save a ton of work and allow for longer term storage, kinda like the CD did when it replaced the floppy disk, but alas, CD's aren't foolproof either.

I will always use slants, but I can't help but be a little adventurous and try the alternative methods.  I appreciate your offer and would be very interested in studying the Kish paper and others.  I would also be interested in your opinion as to the nature of the failures you experienced using the sterile DW method.


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--------------
"Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin

"Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: OICU812]
    #16762705 -

I looked at my collections of pdf papers on the subject of sterile distilled water culture storage, and realized that I had offered too much. All the papers from the Journal "Mycologia" were acquired under a JSTOR academic license, and I am not free to share any copies. A few others were purchased from the academic journals in which they appeared, and also cannot be shared. The rest can be found on the Internet and downloaded.

So what I will do over the next few days is compile a bibliography of all the papers I have on this subject. I'll post it on this thread. I think it may be generally useful to have such a list available.

- Terry


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Liberté, égalité, humidité.

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: Terry M]
    #16763081 -

It took me less time to compile this than I thought. But I'm sure there are some typos, which I will go back and correct. I have copies of all of these papers, and have read them.


Benedek, T. 1961. On Castellani's "water cultures" and Benedek's "mycotheca" in chlorallactophenol. Fragmenta Mycologica 22(7): 255-260.

Borman, A.M., A. Szekely, C. K. Campbell, and E. M. Johnson. 2006. Evaluation of the viability of pathogenic filamentous fungi after prolonged storage in sterile water and review of recent published studies on storage methods. Mycopathologia 161: 361–368.

Burdsall, H.H., and E. B. Dorworth. 1994. Preserving cultures of wood-decaying Basidiomycotina using sterile distilled water in cryovials. Mycologia, 86(2), 275-280.

de Capriles, C.H., S. Mata, and M. Middelveen. 1989. Preservation of fungi in water (Castellani): 20 years. Mycopathogia 106: 73-79.

de Capriles, C.H., S.M. Essayag, A. Lander, and R. Camacho. 1993. Experimental pathogenicity of Sporothrix schenckii preserved in water.  Mycopathologia 122: 129-133.

Castellani, A. 1963. Further researches on the long viability and growth of many pathogenic fungi and some bacteria in sterile distilled water. Mycopathol. et Mycol. Appl. 20, 1-2.

Castellani, A. 1964, The “water cultivation” of pathogenic fungi. Ann. Soc. Belge Med. Trop. 44(2), 217-220.

Ellis, J.J.  1979. Preserving fungus strains in sterile water.  Mycologia 71(5), 1072-1075.

Iacobellist, N.S., and J E. DeVay. 1986. Long-term storage of plant-pathogenic bacteria in sterile distilled water. Applied and Environmental Microbiology, 52(2),388-389.

Mcginnis, M.R., A.A. Padhye, and L. Ajello. 1974. Storage of stock cultures of filamentous fungi, yeasts and some aerobic actinomycetes in sterile distilled water. 28(2): 218-222.

Richter, D.L., and J.N. Bruhn. 1989. Revival of saprotrophic and mycorrhizal basidiomycete cultures from cold storage in sterile water. Can. J. Microbiol. 35: 1055-1060.

Richter, D.L., L.C. Kangas, J. K. Smith, and P. E. Laks. 2010. Comparison of effectiveness of wood decay fungi maintained by annual subculture on agar and stored in sterile water for 18 years. Can. J. Microbiol. 56: 268–271.


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Liberté, égalité, humidité.

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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: OICU812]
    #16764234 -

OICU812 said:
I am especially interested in his maintenance media formula of 4° Balling & 1.5% agar which equates to a 1.016 SG wort + 15g agar per liter.  My calculations show 41.4g of malt extract per liter which seems extremely high to me but I'll break out my brewing refractometer tomorrow (yes, I have been a home brewer for over seven years) and verify the amount of malt extract by weight per liter to equate to 1.016 SG (4°Balling).



I will say from experience that high amounts of malt extract in solution has some issues when being PC'ed Even at 5psi it tends to caramelize and create crystallize. This, at least in liquid cultures and some high solution agars, makes it hard to spot contaminates.

However, that is my findings and could be impurities in the malt extract I buy. I am using brewing grade and not lab grade.


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Re: How do YOU store your samples? OR Interesting Storage Examples [Re: Cyber]
    #16774881 -

Regarding Mycologia articles from the issues 1 to 100, they can be viewed for free at Cyberliber.
http://www.cybertruffle.org.uk/cyberliber/
Just click on 'Journals' in the main menu and choose 'Mycologia'.

For example Ellis, J.J.  1979. Preserving fungus strains in sterile water.  Mycologia 71(5), 1072-1075
http://www.cybertruffle.org.uk/cyberliber/59350/0071/005/1072.htm

Carsten

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