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Offlineg3n3h4x0r
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Mycelium Markers
    #19294664 - 12/18/13 06:46 PM (10 years, 1 month ago)

Hello all!  :grin:

Quick question.  I'm doing some mycology reading and have a few unanswered questions.  However, the answers I'm looking for are more quantitative, possibly species-specific.  So...

Are there any markers, dyes, etc. that can be used to visualize flow through mycelium? 

I'd like to grow mycelium in columns and pipes and pump visualizing agents while varying a few parameters.  I'd like to capture periodic photos during the process for doing measurements and mathematics, so I'll need something that can be seen in the visual spectrum, standard - intense fluorescent (I have a high powered light for visualizing radioactive compounds).

Thanks everyone!  :amanita2:


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OfflineNakor420
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Re: Mycelium Markers [Re: g3n3h4x0r]
    #19298658 - 12/19/13 04:12 PM (10 years, 1 month ago)

radioactive carbon...


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Offlineg3n3h4x0r
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Re: Mycelium Markers [Re: Nakor420]
    #19299408 - 12/19/13 06:49 PM (10 years, 1 month ago)

Quote:

radioactive carbon...




Wouldn't radioactive carbon be sequestered by the mycelium?  I'm trying to visualize flow gradients, so it would need to be something that would not be used, but be permeable.  I could be wrong, though.  Thanks!


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OfflineNakor420
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Re: Mycelium Markers [Re: g3n3h4x0r]
    #19301502 - 12/20/13 05:58 AM (10 years, 1 month ago)

No...psilocybes don't hyper accumulate radioactive carbon as far as I know. I suppose it's possible but I know radioactive carbon has been used as a marker to track nutrients moving through a mycelial network...


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Edited by Nakor420 (12/20/13 05:59 AM)


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Offlineg3n3h4x0r
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Re: Mycelium Markers [Re: Nakor420]
    #19303040 - 12/20/13 01:54 PM (10 years, 1 month ago)

Quote:

No...psilocybes don't hyper accumulate radioactive carbon as far as I know. I suppose it's possible but I know radioactive carbon has been used as a marker to track nutrients moving through a mycelial network...




Interesting - I'll take a look into it.  I think I can get access to some C-14 marked reagents, not sure about pure carbon, though.  Just gotta do some browsing, I guess.  Thanks for the help!


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OfflineNakor420
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Re: Mycelium Markers [Re: g3n3h4x0r]
    #19306605 - 12/21/13 06:46 AM (10 years, 1 month ago)

Np...just for the record...I'm only relaying info I saw in a Paul Stamets lecture,lol.


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Invisiblefastfred
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Re: Mycelium Markers [Re: Nakor420]
    #19310161 - 12/22/13 01:03 AM (10 years, 1 month ago)

There are various stains you could use, but those would permanently stain the tissue and might not be good for the myc.  This would throw off your results if they harmed the myc.

I'd use simple food coloring.  It's cheap, easily available, and unlikely to harm the myc.

Be sure to post pics!  It will be interesting to see how interconnected the network really is.  Depending on the mating types involved you might find patches of unconnected myc which wouldn't take up the dye.


-FF


--------------------
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OfflineNakor420
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Re: Mycelium Markers [Re: fastfred]
    #19310658 - 12/22/13 05:50 AM (10 years, 1 month ago)

I would be interested to know why nutrients move in one direction within one cellular pathway and a different direction in another...What determines which direction chemicals flow through a mycelial thread?


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OfflineAmanita virosa
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Re: Mycelium Markers [Re: fastfred]
    #19317279 - 12/23/13 05:23 PM (10 years, 1 month ago)

Quote:

fastfred said:
There are various stains you could use, but those would permanently stain the tissue and might not be good for the myc.  This would throw off your results if they harmed the myc.

I'd use simple food coloring.  It's cheap, easily available, and unlikely to harm the myc.

Be sure to post pics!  It will be interesting to see how interconnected the network really is.  Depending on the mating types involved you might find patches of unconnected myc which wouldn't take up the dye.


-FF



Seems like food coloring is going to stain the shit out of the chitin in the cell wall and then ur not going to be able to visualize the cytoplasm very well. I would try an oil base stain like Sudan red,
Which won't stain the polysaccharides in the cell wall. But it could be toxic as was earlier stated.  radioactive barium or iodine if you can get some?  Both prolly cytotoxic?  Maybe schedule an upper gi and then bolt with the baruim milkshake. :cool:


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Offlinedrake89
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Re: Mycelium Markers [Re: Amanita virosa]
    #19318718 - 12/23/13 09:59 PM (10 years, 1 month ago)

can anyone find that paper by the team from university of tennessee, where they used radioactive markers to track nutrients through mycorhizae and the associated trees?  i feel like it should be famous but I could never find it...


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Invisiblefastfred
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Re: Mycelium Markers [Re: Amanita virosa]
    #19319642 - 12/24/13 03:56 AM (10 years, 1 month ago)

Quote:

Amanita virosa said:
Seems like food coloring is going to stain the shit out of the chitin in the cell wall and then ur not going to be able to visualize the cytoplasm very well.




Why would you think that?  Food coloring is water soluble.  I don't think it will stain anything.

It should just continue to move with the flow of water.  If it does stain or accumulate in any area it will eventually be redissolved and move along with the flow.


-FF


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OfflineRogerRabbitM
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Re: Mycelium Markers [Re: fastfred]
    #19319844 - 12/24/13 06:40 AM (10 years, 1 month ago)

Tom Volk has an excellent video showing 'flow' through mycelium including the clamp connections.  I saw it in 2005 at a medicinal mushroom conference.  He had some sort of marker, but I can't remember what it was.  Perhaps his video is available somewhere.  Last I heard, he was still at UW Madison.
RR


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OfflineMrcloudy
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Re: Mycelium Markers [Re: RogerRabbit]
    #19320210 - 12/24/13 09:12 AM (10 years, 1 month ago)

Food coloring will eventually be consumed by the mycelium. I did a few tests and dropped several drops onto a plate of reishi, certain colors lasted longer but in the end the color did not really move through the mycelium, although it did stain the agar underneath. Red #40 lasted the longest but after a month or so only a light pink splotch remains.  I have thought about experimenting with synthetic clothing dyes but haven't gotten around to it yet.


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

10 different Ganoderma species from across the USA

AMU

MrCloudys guide to North American GanodermaUpdated A rough guide to North American Ganoderma species, with an emphasis on the laccate species.


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InvisibleWillSolvem
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Re: Mycelium Markers [Re: Mrcloudy]
    #19333647 - 12/27/13 04:59 PM (10 years, 1 month ago)

lycopene or other carotenoids(?)


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AMU Q&A Thread because questions deserve answers.


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OfflineMrcloudy
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Re: Mycelium Markers [Re: WillSolvem]
    #19334620 - 12/27/13 09:35 PM (10 years, 1 month ago)

Apparently Red #40 is a sodium salt. It seems pretty resistant to consumption by the mycelium but still hasn't traveled from the spot it was dropped on. Perhaps if it were dropped on the leading edge of the mycelium instead of on the mycelial mat.


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

10 different Ganoderma species from across the USA

AMU

MrCloudys guide to North American GanodermaUpdated A rough guide to North American Ganoderma species, with an emphasis on the laccate species.


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Invisiblefastfred
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Re: Mycelium Markers [Re: Mrcloudy]
    #19335427 - 12/28/13 03:00 AM (10 years, 1 month ago)

Quote:

Mrcloudy said:
Apparently Red #40 is a sodium salt. It seems pretty resistant to consumption by the mycelium but still hasn't traveled from the spot it was dropped on. Perhaps if it were dropped on the leading edge of the mycelium instead of on the mycelial mat.




You're not going to see any flow if there is no flow.

If all the cells are taking up what they need then there should be no flow.  The only time I'd expect to see any flow is when there's a higher water concentration in one area and a low concentration in another area.


-FF


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Offlineg3n3h4x0r
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Re: Mycelium Markers [Re: fastfred]
    #19342489 - 12/29/13 04:50 PM (10 years, 1 month ago)

Quote:

Food coloring will eventually be consumed by the mycelium. I did a few tests and dropped several drops onto a plate of reishi, certain colors lasted longer but in the end the color did not really move through the mycelium, although it did stain the agar underneath. Red #40 lasted the longest but after a month or so only a light pink splotch remains.  I have thought about experimenting with synthetic clothing dyes but haven't gotten around to it yet.




Great!  Thanks for the info - let me know if you get to it before I do - I'll do the same.

Quote:

The only time I'd expect to see any flow is when there's a higher water concentration in one area and a low concentration in another area.




This is what I'm interested in testing. 

Quote:

Tom Volk has an excellent video showing 'flow' through mycelium including the clamp connections.  I saw it in 2005 at a medicinal mushroom conference.  He had some sort of marker, but I can't remember what it was.  Perhaps his video is available somewhere.  Last I heard, he was still at UW Madison.




I've watched some of his videos through, but can't find the particular one.  I'll keep watching, though!  Still have plenty to learn about mycology - about a third of the way through a text book.


Thanks for the input, everyone!  I'll post my results when I get everything running for anyone interested.

I don't really plan on fruiting any of the tests - more interested in mycelium for my studies, but I'm thinking of using a seed-based medium.  I'm thinking a BFR or grain medium might be too absorbent and might skew my results.  I plan on testing gravity dependence of flow and any ability for a chemical to flow along piping.  Possibly transfer the mycelium to my clear testing container and allowing it to further colonize/consolidate in the dark, then running the tests.  Any thoughts/concerns?


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Invisiblefastfred
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Re: Mycelium Markers [Re: g3n3h4x0r]
    #19353620 - 01/01/14 03:19 AM (10 years, 30 days ago)

Quote:

g3n3h4x0r said:
Quote:

Tom Volk has an excellent video showing 'flow' through mycelium including the clamp connections.  I saw it in 2005 at a medicinal mushroom conference.  He had some sort of marker, but I can't remember what it was.  Perhaps his video is available somewhere.  Last I heard, he was still at UW Madison.




I've watched some of his videos through, but can't find the particular one.  I'll keep watching, though!  Still have plenty to learn about mycology - about a third of the way through a text book.




Just email him.  He's a nice enough guy.  I'm sure he wouldn't mind answering the question.


-FF


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Offlineg3n3h4x0r
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Re: Mycelium Markers [Re: fastfred]
    #19355538 - 01/01/14 05:20 PM (10 years, 29 days ago)

Quote:

Just email him.  He's a nice enough guy.  I'm sure he wouldn't mind answering the question.




Didn't even think about that, I'll give it a try!


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Invisiblelipa

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Re: Mycelium Markers [Re: g3n3h4x0r]
    #19356185 - 01/01/14 08:41 PM (10 years, 29 days ago)

Quote:

g3n3h4x0r said:
Quote:

Just email him.  He's a nice enough guy.  I'm sure he wouldn't mind answering the question.




Didn't even think about that, I'll give it a try!




Please let us know what he comes up with. Yes, he is very nice and has answered every email I have ever written him.


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InvisibleAleon
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Re: Mycelium Markers [Re: fastfred]
    #19357427 - 01/02/14 06:31 AM (10 years, 29 days ago)

Quote:

fastfred said:
Quote:

g3n3h4x0r said:
Quote:

Tom Volk has an excellent video showing 'flow' through mycelium including the clamp connections.  I saw it in 2005 at a medicinal mushroom conference.  He had some sort of marker, but I can't remember what it was.  Perhaps his video is available somewhere.  Last I heard, he was still at UW Madison.




I've watched some of his videos through, but can't find the particular one.  I'll keep watching, though!  Still have plenty to learn about mycology - about a third of the way through a text book.




Just email him.  He's a nice enough guy.  I'm sure he wouldn't mind answering the question.


-FF




He hasn't been at UW at Madison for years.  He's at UW lacrosse, which is quite a distance from madison.  Also many people keep saying UW at Madison is a good school for mycology, but actually it blows.  I have a friend who's a student there wanting to be a mycologist and they only offer 1 or 2 classes taught by a plant pathology professor; they don't even have a mycologist on staff.


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InvisibleMushmitch
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Re: Mycelium Markers [Re: Aleon]
    #19397351 - 01/10/14 05:09 AM (10 years, 21 days ago)

29 mins into this is some info on dying tissues,, http://www.learnerstv.com/video/Free-video-Lecture-1077-Medical.htm

dunno if this is helpful to what you are doing..


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OfflineTrichodermapotamus
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Re: Mycelium Markers [Re: g3n3h4x0r]
    #20366397 - 08/02/14 08:13 PM (9 years, 5 months ago)

Did anything ever come of this?
What markers have you ended up deciding to use for your experiments?


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InvisibleWillSolvem
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Re: Mycelium Markers [Re: Trichodermapotamus]
    #20366550 - 08/02/14 08:58 PM (9 years, 5 months ago)

Quote:

Trichodermapotamus said:
Did anything ever come of this?
What markers have you ended up deciding to use for your experiments?




In my case dichlorofluorescein







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OfflineTrichodermapotamus
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Re: Mycelium Markers [Re: WillSolvem]
    #20387122 - 08/07/14 09:30 AM (9 years, 5 months ago)

Would you be willing to point us towards a protocol you've been using and what you're using it to assess and measure?
Cool stuff.


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OfflineMrcloudy
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Re: Mycelium Markers [Re: Trichodermapotamus]
    #20387164 - 08/07/14 09:44 AM (9 years, 5 months ago)

It looks like as the mycelium grows out it consumes the fluorescence leaving depigmented agar, is this correct?


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

10 different Ganoderma species from across the USA

AMU

MrCloudys guide to North American GanodermaUpdated A rough guide to North American Ganoderma species, with an emphasis on the laccate species.


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