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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Isolating individual spores with a dental pick * 11
    #29697514 -

This is not supposed to work but god damnit we did it and I have receipts.

Background: My KSSS is sporeless, which is annoying because I picked it up specifically to have an obvious pheno to cross against. I've been looking at the gill tissue in the scope periodically to see if more spores develop as the fruits age. So far, no.


This darkfield shot has approx one visible spore near the center, and the rest is bits of cell damage bruising blue. This matters because the following videos are kinda blurry, and it looks like there are a lot of spores at a glance, but there aren't.


Aside: when there are spores, they frequently show up in clusters of 4, sometimes in some perfect clover arrangements, so it seems like spore production might be all-or-nothing on a per basidia basis with these.

--

The one nice thing about this situation is that it naturally dilutes the spore density by like 99%. The idea of "snag a single spore on a needle" is usually a bongrip-tier idea that betrays a lack of appreciation for just how dense a single visible spec of spore already is, how much they want to stick to each other, etc. But in this case they're infrequent enough to actually have a chance.

So I've been trying to actually do it. What ended up feeling best has been a dental pick, deburred with a razorblade to be very finely pointed. It's fine enough to be accurate-ish while being substantial enough to to dig into the plate, grip well, not deform, etc. Needle was too fiddly.

To keep the tissue in place, I choke up on it with a second glass slide and hold it down so it can be prodded without sliding around everywhere.

Sorry this video is such shit. The focus point for the camera and the eyepieces are sliiiiightly misaligned, so having perfect focus in the eyepiece means the video kinda sucks. IRL doing this work was as visible as the photos above.


Managed to whack some loose right at the edge.


Yoink. It looks like I grabbed two here, there's one right on the surface line of the water that looks like it gets picked up first. Did that happen, or did it get pushed back onto the tissue / stay on the surface tension boundary? No idea. Impossible to tell from the video and I was super locked in on the one spore IRL.

As soon as I saw that it got picked up, I immediately jumped into the hood and lightly streaked it on a plate. I then took the wrapped plate back under the scope to try and find whether there was a visible spore in the streak.



I didn't really expect to find anything, but I'll be fucked. Right at the very beginning of the drag where the hook first pierces/touches the agar, there is a single spore and what looks like a tiny bit of coir debris (also visible in the videos โ€“ there was some coir on the original gill tissue). There's nothing else like this visible along the rest of the streak.

--

Will this go anywhere? IDK. The tissue was mounted on the slide with non-sterilized tap water and inspected in open air. The plating is the only thing that happened in the hood. Who knows what other fuckery is along for the ride on that streak. Will the spore even germinate? If it does, will the resulting mono be able to fight off whatever else pops off? No idea.

But that's a single mf spore poached from gill tissue to agar so I'm chuffed :lol:


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.~`  ๐”… ๐”ž ๐”ค ๐”ข ๐”ฉ  ๐Ÿฅฏ  ๐”… ๐”ฆ ๐”ฑ ๐”ข ๐”ฐ  `~.
:salute: CAKE WARS III HAPPENING NOW :salute:

๐Ÿงช Castellani Preservation | ๐Ÿ”ฌ The Slidewich | โ˜ ๏ธ Touchless skull sterilizer | ๐Ÿ“ธ Agar plate scanner


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Re: Isolating individual spores with a dental pick [Re: Rasmodius]
    #29697547 -

You should start making art on grains of rice lol

Super sick write-up Rasmo!! I thought the pics and vids were very detailed :chefskiss:


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๐Ÿ…ƒ ๐Ÿ„ด ๐Ÿ„ฐ ๐Ÿ„ผ    ๐Ÿ„ฒ ๐Ÿ„ป ๐Ÿ„ธ ๐Ÿ„ฝ ๐Ÿ„ถ ๐Ÿ…† ๐Ÿ… ๐Ÿ„ฐ ๐Ÿ„ฟ

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Re: Isolating individual spores with a dental pick [Re: fontinalis666] * 1
    #29697660 -

Congrats, Ras!

Damn, man โ€” this is quite an achievement.

I was thinking this might actually be useful for you. Look, I usually use Japanese acupuncture needles, mine are 0.16mm (almost like a beard hair), and they come sterile and individually wrapped. They have a coiled handle โ€” really comfortable to hold. Really thin, but there are even thinner ones at 0.12mm.



A pack of 100 needles is super cheap and would last you a good long while.

Hugs :heart:


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"Little shark in da bathtub"

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Re: Isolating individual spores with a dental pick [Re: Belintaf]
    #29697956 -

This is awesome :takingnotes:

Definitely bookmark worthy


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"Why try for high fives,
When my pies have nine lives?"
DUBJUG : H.D. : PLASTIC-MYC : TGE : JOIN CAKE WARS : MODERN FRUITING : MCWA : QDMT : MY FAV TEKS ๐Ÿ“–

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