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OfflineAeolus1369
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dH20 vs. an isotonic solution for long term storage
    #4021427 - 04/06/05 11:48 AM (19 years, 1 month ago)

I've read many posts that recommend keeping mycelial cultures in distilled water for long term storage. But have there been any tests done to see if an isotonic solution (one where the concentration of solutes in the solution is equivalent to that inside the mycelium) might be optimal?

Many bacteria and eukaryotic cells will lyse in dH20 because the water diffuses through the cell membrane and generates too much pressure for the membrane to withstand. I know fungi circumvent this because their thick cell-wall can withstand high internal pressure, but perhaps on longer time scales it becomes an issue?

Admittedly this isn't such a huge concern since the "several years" of viability I've seen claimed with just dH20 is probably long enough for practical purposes. Still though, optimization is a recurring theme here so I was just curious if anyone knew anything about the significance of water activity in the culture solution.

--Aeolus

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OfflineSuntzu
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Re: dH20 vs. an isotonic solution for long term storage [Re: Aeolus1369]
    #4025311 - 04/07/05 08:29 AM (19 years, 1 month ago)

I've thought about this a little bit too. The 'experiments' take years to perform, unfortunately.
I think the cell walls should be pretty resistant to osmotic lysing, but I'm not so sure about the wide pH swings that dH2O can go through. Because it's so easy to come by in labs, I've always wanted to try TE buffer; in addition to being somewhat osmotically balanced, it's pH balanced and contains EDTA which curbs rogue enzymatic and bacterial activity. You can use 0.1X TE buffer for many applications as well.
dH2O has worked well for most species I've played with. It seems like the slower growing/finicky species don't survive as well, though. It has never worked for pans, e.g., [IME]

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OfflineAeolus1369
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Re: dH20 vs. an isotonic solution for long term storage [Re: Suntzu]
    #4025820 - 04/07/05 11:09 AM (19 years, 1 month ago)

Hmm, interesting. TE uses Tris as a buffer right? I was thinking the other day how a strong buffer like tris might be used in a casing to ensure long term pH balance but I have no idea if it has any negative impact on mycelium. I would imagine EDTA is to be avoided because it would render many of the mycelium's secreted enzymes inactive but I think it would be beneficial in a long term storage solution as you said.

How does dH20 in a sealed container undergo wide pH swings?

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OfflineSuntzu
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Re: dH20 vs. an isotonic solution for long term storage [Re: Aeolus1369]
    #4031936 - 04/08/05 05:49 PM (19 years, 1 month ago)

dH2O straight from the source doesn't necessarily have a pH of 7. It depends on a lot of factors. If left to equilibrate it usually settles around pH 5.0-6. I'd imagine even a little biological activity could drop it furthur; maybe a buffering agent could keep it constant and closer to optimal.
I can't find the source, but somewhere I heard that ultra-pure water ['ddH2O'] can have an initial pH of something like 3-9; Most likely any water will settle to 5-6 over a short period of time, though.

So I wonder what 'optimal' really would be?

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