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Mushrooms, Mycology and Psychedelics > Advanced Mycology
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OfflineFishLevelMidnight
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Registered: 09/01/17
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Scloteria long term storage for reanimating tissue
    #24713996 - 10/16/17 11:59 AM (6 years, 5 months ago)

Hey all.

If I take a clean scloteria stone and place it in distilled water, will it keep long term?

I know dH2O can be used to store mycelium for long periods of time; would it be possible for one to store a stone in dH2O at 4°C and then recover myc for inoculation off of agar later?

I am just curious what will be the best/easiest way to keep my galindoi for later use (other than spore print). I will try for prints, but I will also store mycelium in dH2O off agar if the stone method won't work or is untested.

Thanks and happy grows!
Fisherman


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OfflineAlan RockefellerM
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Re: Scloteria long term storage for reanimating tissue [Re: FishLevelMidnight]
    #24718605 - 10/18/17 03:26 AM (6 years, 5 months ago)

That sounds like a good way to do it.

Also try storing stones in a vial without the water.

Also try making a culture slant, and instead of inoculating it with an agar wedge, drop a sterile stone on it.

Freeze drying is what the professionals use, but the equipment is quite expensive.

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Offlinekrypto2000
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Re: Scloteria long term storage for reanimating tissue [Re: Alan Rockefeller]
    #24718906 - 10/18/17 08:41 AM (6 years, 5 months ago)

Slants are generally how any long term cultures are preserved afaik. Even freeze drying/storing cryogenically I believe that's true, but not positive on that one.

edit: I currently do not have slants... actually I bought some but lost the test tubes and rack, so I just keep agar dishes in the fridge. Every 6-12 months I'll pull them out and refresh them (eg transfer to a new dish and let it grow out). So far that has worked for me for 2-3+ years and although I've had some that were very slow to recover I have not yet lost a culture. The ones that are slow to recover I probably waited over a year too though.

Edited by krypto2000 (10/18/17 08:43 AM)

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InvisibleAndyHinton


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Re: Scloteria long term storage for reanimating tissue [Re: FishLevelMidnight]
    #24984598 - 02/11/18 12:00 AM (6 years, 2 months ago)

It's interesting that you mention sclerotia shavings in water, I've seen that with Perigord truffles in tiny vials. For bitter stones we need honey. :smile:

I learned the hard way that dH2O kills most fungi within a year due to osmotic pressure imbalances. Water cultures progress slowly and quietly. Just like how people remembered Castellani's 1939 research much later, so had Benedik already solved the problem with saline in 1962.

Water cultures fill an important niche between keeping slants for daily use and making cryovials because when refrigerated, they outperform anything else you can store in a home unit with auto-defrost. You can also keep them at room temperature and back up your collection, e.g., in a safe deposit box for up to 5 years.

My current technique is based on blood serum and involves mycelium suspended in 0.6% sea salt–dH2O saline, stored in silica-coated BD Vacutainer serum tubes (red top). The 33% salinity reduction and basal salts are based on Awad et al., "The history of 0.9% saline"; and Tresner and Hayes, "Sodium Chloride Tolerance of Terrestrial Fungi."

Normal 0.9% saline is arbitrary or derived from an osmotic coefficient calculation specific to human blood (0.6% salt). 50% of the salt-tested fungi died in a 2% concentration. 6 g/L sea salt in dH2O seems to be where it's at for an inert medium. It's also a good hangover cure. :smile:

PS. I was thinking about the LC-soaked perlite cryovials and wondered if a Sabouraud-type mixture with peptone, extra potato dextrose, and 10% glycerol might be the best formulation for that purpose. My LB is already made up and I want to add gentamicin later.


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Edited by AndyHinton (02/11/18 12:06 AM)

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Mushrooms, Mycology and Psychedelics > Advanced Mycology
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