Resinomycena saccharifera - difficulties with microscopy

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Steve
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Resinomycena saccharifera - difficulties with microscopy

Post by Steve » Sun Jul 26, 2015 9:29 pm

Hi,
I was "hitting the wall" with my microscopy manipulation skills with this one - it was so tiny and flexible that I was quite unable to cut off gills, slice cap cuticle or do anything apart from cut of bits and make a squash :(
I'm fairly sure this has to be Resinomycena saccharifera – a small white Mycena-like toadstool on marsh plants. John and I found it on the Longshaw Estate on partly submerged plant stems with Juncus, on which Mycena bulbosa was also found. The description in Funga Nordica is the best fit I can find in my books, for overall dimensions, texture, colour: white turning slightly yellow-brown, crenulate cap margin, and gills which are broad and distant. Spores were also a good fit - elongate tear-shaped 9-12 (13) x 3-4.5. Reaction with iodine was not easy to see as the spore mass was so minute, and the slight reaction noted in Funga Nordica may not be obvious on single spores (especially in blueish LED light). However, the dextrinoid reaction of the cortex was more convincing. Basidia were very hard to see once the toadstools had dried and were rehydrated – one single two-sterigmata basidium was micrographed. The tiny toadstool was basically impossible for me to section, or dissect in any way – I had to make squashes of chunks, meaning that I was unsure which structures belonged to which bits. However, nothing seems to instantly rule out Resinomycena saccharifera, I think.
I have 4 local records, 2 of which look like replicates from a 2011 BMS foray to Worsborough (anonymous but presumably trustworthy), and two from the Longshaw Estate in the 1980’s, one anonymous, and one from Tony Lyon which can be trusted. So it's not something unexpected in our area.
Steve
Attachments
Meltzers.jpg
To test for a dextrinoid reaction I dunked a whole toadstool into Melzers - seen lit from below (left) and above (right). The orange colour is less obvious in the blueish LED light of my microscope, which supresses red.
Microscopy.jpg
Clamps top left, cystidia, cap cuticle bottom right perhaps...
Spores.jpg
Spores in water, oil immersion
Macro.jpg
Stereo microscope images

Steve
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Re: Resinomycena saccharifera - difficulties with microscopy

Post by Steve » Mon Jul 27, 2015 9:20 pm

Hi,
My post maybe looks scary as no-one is responding. It's mean to be a starter to a discussion on microscopy of tiny fungi. There must be lots of expert microscopists out there who can offer tips and advice.

How to cut a section of a tiny fungus :?:
Required:
Razor blade (37 mm) to cut section
Dissecting needle (60 x 0.5 mm) to hold specimen and pick up section
Small drop of water on microscope slide for section
Cover slip
Binocular (stereo) dissecting microscope
Put specimen e.g. an Unguicularia cup fungus of 0.2 mm diameter under microscope.
Hold with dissecting needle and cut off a 20 um slice with the razor blade.
Use the dissecting needle to transfer to the drop of water.
Place cover slip on drop of water.

Try doing this at x45 under the stereo microscope. A virtual reality where everything looks 45 times bigger. You yourself are now over 80 yards high.
You feel as though you are in a giant’s kitchen and are about to slice a pea: this one is about a third of an inch across (0.2mm x 45 = 9 mm).
The knife (razor blade) you are using is five and a half feet long.
You hold the pea steady with a sharpened pole (ie the dissecting needle) 9 feet long.
Then you take the knife between your finger (which is 12 feet long and 3 feet thick), and your thumb (which is 10 ft x 4 ft).
The slice should be something like ONE MILLIMETRE thick in your x45 world (ie 20 microns x 45).
You pick up this wafer thin slice of pea with the 9 foot pointy pole. (A touch of the DTs, or a minute awkward flick and everything is hurled a vast distance and lost. Some sliced dried material simply whizzes off to infinity like a highly charged particle).
With luck you place the slice in a drop of water on a slide which is 11 ft x 3’ 6”. (That’s if the slice doesn’t stick like glue to the pole end and won’t wash off into the water).
On top of this goes a cover slip, a yard square. Where is that slice of pea?
Of course you can use a lower stereomicroscope power – but it’s a tiny fungus...
Attachments
Cutting a section.jpg
All objects to same scale at x45 (including finger)
Tools.jpg
Tools for the job
Hyalopeziza millepunctata (Unguicularia scrupulosa).JPG
A tiny fungus (Hyalopeziza millepuncta perhaps) x45

roy betts
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Re: Resinomycena saccharifera - difficulties with microscopy

Post by roy betts » Tue Jul 28, 2015 8:55 am

No need to reply - you seemed to have it sorted! Except the spore width is a little narrow and they're usually 4-spored.
Sometimes I find you can see if small specimens are 2 or 4-spored using low power and looking directly up at the gills (gill cystidia also often visible; if present). Otherwise squashing half a fruitbody is the only way. This is OK if you know what you're expecting to see (eg: Mycena adscendens has distinctive cheilocystida and cap veil cells).
Fortunately I'm not addicted to the 'hard stuff' (brackets, crusts) so never have cause to examine these microscopically!

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Lancashire Lad
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Re: Resinomycena saccharifera - difficulties with microscopy

Post by Lancashire Lad » Thu Jul 30, 2015 12:44 pm

I originally typed this response: - viewtopic.php?f=5&t=336 (relating to a method I use for preparing tiny samples) here, but moved it into "Tips & Techniques", as it may be more visible there for future searches.

Regards,
Mike.
Common sense is not so common.

Steve
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Re: Resinomycena saccharifera - difficulties with microscopy

Post by Steve » Sat Aug 01, 2015 4:47 pm

Many thanks Roy and Mike,
Some good tips!
Sorry for tardy response - a full day at Longshaw on Weds means a long time on the microscope.
I find hypodermic needles extremely useful for picking up whole fruit bodies (as you say - sometimes it's the only way to get a specimen onto the slide). They are also very useful for dealing with tiny amounts of water and reagents. I'm not 100% sure about the legal situation of hypodermic needles but you can get them on Amazon. I suspect that they are only illegal as part of a drug-kit.
Regards,
Steve
Attachments
3.JPG
Picking up an Unguicularia fruit-body
2.JPG
At x 45 the Unguicularia is still mighty small
1a.JPG
The fine hypodermic in action. The "big" mushroom is Crepidotus epibyrus. The Unguicularia is almost invisibly small.

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