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What's up with these Petri dishes, what's this pattern? * 2
    #29079157 -

Hey there, I've been struggling since months in trying to obtain some clean mycelium. The following dishes are representative of what I'm getting at the moment.



What's your opinion on these? Especially, how should I judge this weird circular pattern? Is this symptom of some kind of contamination? I'm trying to get better at figuring out what's happening on my transfers, so I'd appreciate the reasoning. FWIW the smell is good, smells like mushrooms.

They come from a GT clone (which in turn came from a spore print), then after several transfers I managed to obtain the above. I never had any particular contamination AFAIK, but I struggled very hard to get rid of what looks to me as a second culture, which in most cases also seemed to grow on top of the rhizomorphic mycelium in the form of a more tomentose one. Is this even possible? By the way, see an example below.



Any help would be greatly appreciated. Thank you folks!


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Re: What's up with these Petri dishes, what's this pattern? [Re: jee3apha] * 2
    #29079228 -

Very pretty plates, nice work!  They look good to me, but I’m only a pretend mycologist. :wink:  I see some typical sectoring, which can come from different strains on the same plate - such as between 5&7 o’clock on the lower plate in your first pic, but it also looks like a small bit of transferred agar/material may have gotten rolled from center a bit.  In any case, looks really pretty to me….certainly good enough for grain. No bacteria or mold to my aged eyes. 

If you are worried about something hiding on that plate, one approach that can work well is described in the following paper, linked as a PDF: https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0224635&type=printable

Good luck!


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Re: What's up with these Petri dishes, what's this pattern? [Re: jee3apha] * 1
    #29079304 -

:whathesaid:

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Re: What's up with these Petri dishes, what's this pattern? [Re: Agar Wedge] * 2
    #29079312 -

Beautiful plates!  I'm a hobbyist, not a pro but perhaps the ring you are referring to was created due to a fluctuation in temps that caused a burst of growth?  I've seen some of my plates get the look of tree rings if I moved them around a lot.


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Re: What's up with these Petri dishes, what's this pattern? [Re: Quailfishbeach]
    #29079503 -

:whathesaid:

What is the temperature difference between day and night in the room?

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Re: What's up with these Petri dishes, what's this pattern? [Re: flapping_bird]
    #29079681 -

flapping_bird said:
Very pretty plates, nice work!  They look good to me, but I’m only a pretend mycologist. :wink:  I see some typical sectoring, which can come from different strains on the same plate - such as between 5&7 o’clock on the lower plate in your first pic, but it also looks like a small bit of transferred agar/material may have gotten rolled from center a bit.  In any case, looks really pretty to me….certainly good enough for grain. No bacteria or mold to my aged eyes. 



Thanks! I always considered uneven culture, a sign that something's wrong, I mean in most cases it is, I'm just not experienced enough to understand the nuances.

For example, I was worried that in the middle of the plates the mycelium is way thinner than on the outside, and I've found that's not the case in one other plate of the same batch (not shown in the pictures) in which it is way more uniform.

Quote:
If you are worried about something hiding on that plate, one approach that can work well is described in the following paper, linked as a PDF: https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0224635&type=printable



Wow this is extremely cool. Thank you! <3


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Re: What's up with these Petri dishes, what's this pattern? [Re: fungusul]
    #29079682 -

Thank you!

Quailbeach said:
I'm a hobbyist, not a pro but perhaps the ring you are referring to was created due to a fluctuation in temps that caused a burst of growth?  I've seen some of my plates get the look of tree rings if I moved them around a lot.


fungusul said:
What is the temperature difference between day and night in the room?


There is no temperature difference or fluctuation, since the culture was about 1 inch of diameter, I moved the plates into the incubator, which kept a pretty much steady temperature of 72.5 °F (22.5 °C).


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Re: What's up with these Petri dishes, what's this pattern? [Re: flapping_bird]
    #29081121 -

flapping_bird said:
If you are worried about something hiding on that plate, one approach that can work well is described in the following paper, linked as a PDF: https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0224635&type=printable



Have you tried this personally? Any practical advises? I'm going to give it a try tomorrow with some spare backup plates.


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Re: What's up with these Petri dishes, what's this pattern? [Re: jee3apha]
    #29096570 -

jee3apha said:
flapping_bird said:
If you are worried about something hiding on that plate, one approach that can work well is described in the following paper, linked as a PDF: https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0224635&type=printable



Have you tried this personally? Any practical advises? I'm going to give it a try tomorrow with some spare backup plates.


So I tried it, and this is the result, I'm kind of surprised that 7 dishes out of 7 basically succeeded, at least I think so, here's what it looks like after the transfer.



I could add some more details if someone's interested.


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Re: What's up with these Petri dishes, what's this pattern? [Re: jee3apha]
    #29096574 -

jee3apha said:
jee3apha said:
flapping_bird said:
If you are worried about something hiding on that plate, one approach that can work well is described in the following paper, linked as a PDF: https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0224635&type=printable



Have you tried this personally? Any practical advises? I'm going to give it a try tomorrow with some spare backup plates.


So I tried it, and this is the result, I'm kind of surprised that 7 dishes out of 7 basically succeeded, at least I think so, here's what it looks like after the transfer.



I could add some more details if someone's interested.



I'd appreciate some extra details.

I tried it once, but found that my donor plate was thicker than my cabin plate. Because of that I wasn't able to get the roof to stay put.

I've poured some thicker plates for the cabin that I haven't tried yet.

I'm also concerned that I won't be able to push the air out. The roof doesn't seem to want to stick, just lay on top of wherever I put it.


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Re: What's up with these Petri dishes, what's this pattern? [Re: PuffM4gicDr4gon]
    #29098140 -

PuffM4gicDr4gon said:
I tried it once, but found that my donor plate was thicker than my cabin plate. Because of that I wasn't able to get the roof to stay put.

I've poured some thicker plates for the cabin that I haven't tried yet.



You definitely want to pour way thicker plates if you do this! If you do so you'll be fairly sure that your transferred agar chunk will fit the cabin.

Quote:
I'm also concerned that I won't be able to push the air out. The roof doesn't seem to want to stick, just lay on top of wherever I put it.


Well if the cabin agar is thick enough to contain the chunk, all this will happen by itself. The agar is completely flat, so is the coverslip, basically you just have to let the coverslip fall onto the cabin, then you can gently tap it, just to be sure. I definitely had zero issues with air. You just need to be very careful when you place the coverslip because I think is basically impossible to move it afterwards, without ruining the agar. It's weird that you say that it doesn't stick, a flat glass surface and agar should stick like glue.

Biggest issue I had was to pick each individual coverslip using a pair of sterilized tweezers; moisture kept them all stuck together. Lesson learned: keep each coverslip separated by something, e.g., a layer of foil.


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