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InvisibleDerGeist
Completing theCycle

Registered: 12/23/04
Posts: 135
Loc: The Other Side
Perhaps not using the right search words..
    #3605408 - 01/09/05 10:49 PM (12 years, 6 months ago)

Well I looked, but wasn't getting quite the answers I was looking for. Say you had a foaf that was wanting to start a mycelium culture in one of the various mediums, one can do this in. How long can you "stall" the growth? Say. Placed in a somewhat cool environment? And still have viable mycelium to work with?

Sorry if this should be a given, but I trudged through tons of How-To posts, but nothing answered this well enough.

DG


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Invisibledog
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Registered: 06/20/02
Posts: 2,790
Loc: Route 66
Re: Perhaps not using the right search words.. [Re: DerGeist]
    #3605437 - 01/09/05 10:59 PM (12 years, 6 months ago)

I've stored colonized and partially colonized substrate jars in the fridge for months. Once they warm up again they grow normally, provided they have not dried out.


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Fascism (fash'izem) n. A governmental system marked by a centralized dictatorship, stringent socioeconomic controls, and often belligerent nationalism. see also: the Bush Administration.


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Invisibleagar
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Registered: 11/21/04
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Trusted Cultivator
Re: Perhaps not using the right search words.. [Re: DerGeist]
    #3605459 - 01/09/05 11:05 PM (12 years, 6 months ago)

First it depends on the method & medium.

You can remove a piece of healthy myc off an agar plate, store it in sterile oxegen free water in a test tube for years.

You can store a well prepared culture on agar, in a fridge for months.

You can store a fully colonized grain / wbs jar in the fridge for longer than a month.

What is the specific medium & storage - you want to accomplish?


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InvisibleDerGeist
Completing theCycle

Registered: 12/23/04
Posts: 135
Loc: The Other Side
Re: Perhaps not using the right search words.. [Re: agar]
    #3605479 - 01/09/05 11:11 PM (12 years, 6 months ago)

This foaf is taking a vacation here shortly, and was hoping to start a culture that would be somewhat developed when they got back. In hopes of getting a jump start on the new hobby they seemed to have been enchanted with..  Now this isn't your typical weekend vacation, mind you, so a longer period of time would be needed. It would need to survive 1.5 to 2.5 months in a fridge... They really don't have a plan as of yet, but researching the options. :smile:

Just Pipe Dreaming?

DG


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OfflineTantalus
Beyonddescription.
Registered: 06/05/03
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Last seen: 11 years, 10 months
Re: Perhaps not using the right search words.. [Re: agar]
    #3605528 - 01/09/05 11:22 PM (12 years, 6 months ago)

Quote:

agar said:
You can remove a piece of healthy myc off an agar plate, store it in sterile oxegen free water in a test tube for years.






Oxygen free water?  Can you explain this one?  I don't think that's possible.  :confused:


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"A nation's hope of lasting peace cannot be firmly based upon any race in armaments but rather upon just relations and honest understanding with all other nations...

Every gun that is made, every warship launched, every rocket fired signifies, in the final sense, a theft from those who hunger and are not fed, those who are cold and are not clothed... The cost of one modern heavy bomber is this: a modern brick school in more than 30 cities. We pay for a single fighter with a half million bushels of wheat. We pay for a single destroyer with new homes that could have housed more than 8,000 people..."

President Dwight Eisenhower, 1953


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Invisibleagar
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Re: Perhaps not using the right search words.. [Re: Tantalus]
    #3605557 - 01/09/05 11:28 PM (12 years, 6 months ago)

I used a poor choice of words. Should have said "sterile water".

Sterile water was first used to preserve cultures by Castellani in 1939 (2). Since then, many scientists have used this method; McGinnis et al. in 1974 (3) and Odds in 1991 (4) reported that they were able to maintain viable cultures for more than three years, without degradation.

This technique satisfies many different interests: Castellani's pathogenic fungi, Odds interest was in pathogenic yeast, and McGinnis' interest was in a wide range of fungi, yeast, and bacteria. The distilled water preserved all of them.

Storage Method:
Obtain dram vials from your laboratory supplier and fill them about half full (about 3mL) with distilled water, loosely cap the vials and sterilize them in a pressure cooker for 30 minutes @250 F. Half dram vials or test tubes with screw caps would also work well.

About six milliliters of sterile distilled water is pipetted aseptically into a freshly growing culture. The fragments of hyphae are dislodged by lightly scraping the aerial growth with the same pipette, and the resulting suspension is withdrawn and transferred to a sterile glass vial. Put plenty of inoculum into each vial to insure success. Screw the lid on tight and wrap Parafilm around the top of the vial to make sure it is airtight. When you come back in a few years, you would not want to find that the water had evaporated
.
Store the vials at room temperature away from direct sunlight. A bookshelf or wall cabinet is an excellent place. If conditions deteriorate, and the room should become unbearably hot, the vials can be refrigerated, but that is not normally necessary.

In the distilled water environment, the mushroom culture enters a dormant state, and it is held in stasis
.
The Rude Awakening:
Under aseptic conditions, simply dip a sterile loop into the vial, and streak the mycelia-rich water onto an agar plate. It will start to reanimate on being in a nutrient source and oxygen.

The first four methods keep cultures alive with three items: food, water, and oxygen. If they lack any of these, It's goodbye.

Instead of trying to keep them alive, there is a better way: In sterile distilled water, with no food, oxygen, or minerals. This method was in use almost 60 years ago, but was apparently lost due to lack of communication.

References
(1) M.D.Graham, "A Simple,Practical Method for Long Term Storage of Yeast", Brewing Techniques 5, March/April (1997), pp 58-62
(2) S.Castellani,"Viability of Some Pathogenic Fungi in Distilled Water", Journal of Tropical Medicine and Hygiene 42, pp 225-226 (1939)
(3) M.R.McGinnis,A.A.Padhye,and L. Ajello,"Storage of Stock Cultures of Filamentous Fungi,Yeasts, and Some Aerobic Actinomycetes in Sterile Distilled Water", Applied Microbiology 28, pp 218-222 (1974)
(4) F.C.Odds, "Long Term Laboratory Preservation of Pathogenic Yeast in Water",Journal of Medical and Veterinary Mycology 29, pp 413-415(1991)


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