JoeTherapist said:

Thanks for the feedback! I will try to get better pictures in front of the FFU. This is T1 from LC. There is no contamination from my untrained eye, but if there is anything riding along, I don’t know what I’m looking for. I would love to be able to try some techniques to clean it up.

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bluescreen said:

Did this cabin (I know it’s not a proper cabin using an agar roof), but the plate was a partial pour, and I love how the myc is reaching out on the virgin plastic.

Reminded me of something Faht mentioned about sticking a piece of agar on the petri lid, and letting the myc reach up to it as a possibility for really cleaning things up. I don't know if it was just an idea or if anyone has tried it, but now I want to.

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Quote:

Bigworm said:

I don't see anything wrong with using an agar roof. I do it all the time and scrape the agar off that grows through it for a transfer.

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Yeah it's officially a cabin with a microscope slide cover slip or similar, just trying to be official. As I understand it

Bigworm said:

I know, but it works either way. The agar roof was the original then the slide cover came into play. At least that's how I took it.

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Yes it looks like it works either way, but on the big thread on cabins some folks pointed out it's a cabin with a slip, and a kind-of cabin with agar. I'm fairly new to all this so I have cover slips for the next ones.  But I'm going to try Faht's suspended ceiling first.

Bigworm said:

Look into cabin plates, and crevis or crevasse plates and water agar.

Water agar is the simplest. Water and agar. That's it. Put your culture in the center and let it grow out. Look for a really faint almost invisible leading edge and take a transfer. That's it.

The cabin plate consists of cutting a small square in the agar and removing it so it's an empty space. Then you take a transfer from the donor plate and put it in the center of that space on the new plate. You cover the space housing the transfer with a slide cover or a piece of what bigger then the cutout. Now the myc will jump off and climb out onto the agar surface, leaving anything piggybacking behind. Good for bacteria may help with mold.

Crevis plate is the same concept but you cut a 3mm wide line of agar out across the width of the plate and create a gap. Place the donor transfer on one side of the gap. My will jump across leaving the contams behind. Also good for bacteria may help with mold. A hot pour plate or brf puck would be good for mold.
I saw something called flap plate where you cut a ^ in the plate and stick a transfer under the flap. The myc will grow through the agar and supposably, you got a clean culture by scraping the stuff that grows threw the agar and sending to another plate.

I do a cabin plate then go to a crevis plate, then send it to water agar, and finally a regular plate. Why? Cause I'm paranoid and I've had a hell of a time with trich for a long time. I think I got it under control now. Water agar is awesome for cleaning up cultures.

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I’m taking a break now from doing the cabin plates with an agar roof. I also threw a crevice in the plates I took the sample from. I had a batch of agar I poured too hot and so there is WAY too much condensation. I am seeing some halo’s around the mycelium but can’t tell if it’s just new growth, moisture, or contamination. I have not yet experienced blatant contamination outside of two plates that never had mycelium. I will do the plates with the tiny halos after they grow out some more. Next time I pour I will make some water agar and low nutrient agar. I have MEA and grain water from boiling oat spawn I want to experiment with too. Thank you for the help! I will post the results of the cabins and trenches soon. I tried getting some pics in front if the FFU but they didn’t turn out well. These feel like they were going strong then thinned out and stalled.

These were the first transfers I ever did on pre-made plates. The darkest blue plate is LC not a transfer. The other two are T1 done in SAB. My own plates look WAY better.

MethodicalMystical said:

I don't really get why 10g nutes/500ml water is the standard recommended ratio around here. IME this resulted in slow growth and, as far as I understand, is about twice as much as microbiologists tend to use in lab settings. So after testing a bit, 6-7g has worked much better for me.

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