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vdlsar
Stranger
Registered: 11/10/07
Posts: 33
Last seen: 11 years, 12 days
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Preparing mushroom tissue for microscope viewing?
#7815525 - 12/30/07 11:20 PM (16 years, 1 month ago) |
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I just got a decent, inexpensive microscope and I've been having fun looking at prepared slides. Does anybody have tips how to prepare and mount various mushroom tissues for viewing? I removed a single gill from a store-bought mushroom and tried a water mount but ended up with bubbles between the slide and the cover. Got a clear and compelling view but the cover slip would not lie flat, the gill was too thick. This rules out use of the 100x oil-immersion lens.
I also have a microtome. It seems like a simple enough tool to use. I tried to freeze and slice a section of oyster mushroom stem, but my freezer can't freeze the tissue solid enough to get a good slice. Would a sharper blade help, or do I just need to freeze it more solidly? I tried mounting a slice anyway, and found that the hyphae are practically invisible. So now I'm wondering what kind of stain to use and how to apply it.
Spores themselves are easy enough to look at. But I'd really like to see mycelial tissue, preferably still alive and growing.
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RogerRabbit
Bans for Pleasure



Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 11 months, 3 days
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Re: Preparing mushroom tissue for microscope viewing? [Re: vdlsar]
#7815560 - 12/30/07 11:34 PM (16 years, 1 month ago) |
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Instead of a slice, try scraping the scalpel so you 'shave' a very small piece. You should be able to get a paper thin curly from your scalpel that you can put a drop of distilled water over and then the cover slip should lay flat. There's mounting solutions if you want a permanent mount, but distilled water works great for immediate viewing. Tap water will have too much crap floating around in it. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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LordByron
Kwisatz Haderach



Registered: 10/07/05
Posts: 33
Loc: Cascadia
Last seen: 5 years, 1 month
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Re: Preparing mushroom tissue for microscope viewing? [Re: RogerRabbit]
#7830318 - 01/03/08 11:47 PM (16 years, 29 days ago) |
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I like to use Melzer reagent to stain tissue but this is very hard to come by unless you know someone in a bio lab. In a pinch you could try a small amount of iodine heavily diluted in distilled water.
Trying to cut a gill section of a mushroom for viewing is an art that takes a lot of practice. You can try my short cut. Take the smallest gill section you can cut, add a drop of what ever mounting solution you can muster, cover with a plastic slip cover, mash flat with a clean pencil eraser. Your subject is now 2d enough for the oil immersion lens.
For live growing tissue under the microscope, you are looking at some seriously fancy hardware.
-------------------- All you touch and all you see, is all your life you'll ever be.
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fastfred
Old Hand



Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
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Re: Preparing mushroom tissue for microscope viewing? [Re: LordByron]
#7830519 - 01/04/08 01:07 AM (16 years, 28 days ago) |
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Just skip the coverslip and put oil right on the specimin. That should work as long as the specimin is pretty flat, like gill tissue.
For viewing live specimins just don't kill them. I look at live stuff growing in petri dishes all the time. If you pour your agar right close to the top of the dish you can look through the cover at 400X pretty easily. Otherwise you can use 1000X oil by taking off the cover and placing the oil directly on the agar.
None of the micro 101 type stains I tried worked very well with myc. Crystal violet, methyl blue, gram stains, etc. didn't really seem to do much for viewability. I didn't try a negative stain though, so I encourage you to experiment. It would be nice to find a good stain to use.
-FF
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