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 gsharpnolack
Registered: 09/05/11
Posts: 657
Loc: Washington State US 
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 Stropharia ambigua cheilocystidia 
#15794363 - 02/11/12 06:44 PM (3 months, 15 days ago) |
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Found in a woodchip trail
Stropharia ambigua - Field image
Underside
Cheilocystidia (no stains - no cover slip)
Cheilocystidia (no stains - no cover slip)
The same section first gets a drop of koh then a drop of congo red (no cover slip)
The same section first gets a drop of koh then a drop of congo red (no cover slip) Image 2
Same section - A sporeless basidium reveals its sterigmata
Same section - Spores within the gill (and I believe the cylinders are chrysocytidia?)
Same section - Spores within the gill #2
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Gravija
Registered: 06/28/11
Posts: 1,766
Loc: Chicago, Illinois, USA
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Re: Stropharia ambigua cheilocystidia [Re: gsharpnolack]
#15794385 - 02/11/12 06:49 PM (3 months, 15 days ago) |
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That is neat!
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Byrain
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Registered: 01/07/10
Posts: 1,819
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Re: Stropharia ambigua cheilocystidia [Re: Gravija]
#15794487 - 02/11/12 07:07 PM (3 months, 15 days ago) |
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Mount it in KOH if you have it and look for chrysocystidia!
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inski
Cortinariologist
Registered: 02/28/06
Posts: 2,565
Loc: New Zealand 
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Re: Stropharia ambigua cheilocystidia [Re: gsharpnolack]
#15794554 - 02/11/12 07:19 PM (3 months, 15 days ago) |
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Nice find, could you explain the reason for not using a cover slip please.
Another interesting microscopic characteristic you could look for is acanthocytes, these cells are found in the basal mycelium of Stropharia spp and in my experience at least one Psilocybe species, they can most likely be found in other Psilocybe species also.
They are generally star shaped or stellate with many pointed projections and are very interesting as it has been shown that they are able to trap and kill nematodes, the nematodes are consumed by the fungus and it is speculated that these fungi are obtaining nitrogen from the trapped nematodes to aid in survival.
Here is a link to a paper on Acanthocytes in Stropharia rugosoannulata and their function as a Nematode-attacking device.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1449000/
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gsharpnolack
Registered: 09/05/11
Posts: 657
Loc: Washington State US
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Re: Stropharia ambigua cheilocystidia [Re: inski]
#15794735 - 02/11/12 07:49 PM (3 months, 15 days ago) |
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With regards to the cover slip, it seems to be a more natural and therefore more authentic approach at microscopy - although not without caveat. I am now realizing the importance and necessity of koh and congo red - which are hardly natural approaches, but they are necessary sometimes.
I'm specifically interested in a mushroom's original appearance - without stains and without the potential of impacting microscopic forms by pressure from a cover slip. Stains, cover slips, crush mounts, etc are of course valid to me and useful but only in comparison to the truest, original form.
Another note, instead of having to deal with angles and concerning one's self with the impossibly thin gill cross section, a preparate cut-out (pictured above) is perfectly flat and allows for a very simple, direct sight. I encourage everyone involved in microscopy to try it the next time you harvest a large mushroom from the forest floor.
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One thing that can improve our planet: Personally creating a small amount of special atmosphere within the home place.
http://en.wikipedia.org/wiki/Four_Noble_Truths
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gsharpnolack
Registered: 09/05/11
Posts: 657
Loc: Washington State US
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Re: Stropharia ambigua cheilocystidia [Re: gsharpnolack]
#15795261 - 02/11/12 09:29 PM (3 months, 15 days ago) |
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Blue-green bruising on cap in areas where it touched black colored heavyweight construction paper
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One thing that can improve our planet: Personally creating a small amount of special atmosphere within the home place.
http://en.wikipedia.org/wiki/Four_Noble_Truths
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RigVedaLXVII.Soma
Bacteri/Phyc/Bry/Myc/ology
Registered: 11/21/11
Posts: 380
Loc: Michigan
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Re: Stropharia ambigua cheilocystidia [Re: gsharpnolack]
#15796170 - 02/12/12 02:51 AM (3 months, 15 days ago) |
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Right on man. Sweet posts as usual.
You mention how you would like to the view the mushroom in its natural look, how about snapping multiple shots at different focuses and focus stacking them? Then you would see everything in 3D like detail to give way more depth of field. It can really show some things you would never notice without doing so. Plus you would see many, instead of seeing just a tunnel vision focus of one object with background defocus.
For example, There is literally no other way to view this image, that I snapped at multiple focuses and focus stacked. Looking at the same thing under the scope in cross section reveals only clear protrusions with no detail.
It is the underside of a thallus of a Lichen.
If you don't know how to focus stack I could do it for you on some images if you would like. The resolution would look better than that last image, I just accidentally moved a little.
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     "While clinging to the moronic belief that they constitute a "counterculture," they share our society's overriding urge for expediency. They make no attempt to learn about the organisms they eat and it always struck me as ironic that people with such a low level of consciousness should be seeking "higher consciousness." -David Arora
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TimmiT
Registered: 03/23/10
Posts: 2,431
Loc: Victoria 
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Re: Stropharia ambigua cheilocystidia [Re: gsharpnolack]
#15796515 - 02/12/12 06:56 AM (3 months, 15 days ago) |
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I'm not sure I understand the natural/authentic approach to microscopy.
I suggest that you always use cover slips... if for no other reason than to protect your objectives from accidental contact with your sample. A cover slip should not change the appearance of microscopic structures.
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"Reality leaves a lot to the imagination" ~ John Lennon
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inski
Cortinariologist
Registered: 02/28/06
Posts: 2,565
Loc: New Zealand
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Re: Stropharia ambigua cheilocystidia [Re: TimmiT]
#15797537 - 02/12/12 12:06 PM (3 months, 15 days ago) |
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Quote:
TimmiT said:
I suggest that you always use cover slips... if for no other reason than to protect your objectives from accidental contact with your sample. A cover slip should not change the appearance of microscopic structures.
Agreed, not only for the reasons stated above, with no cover slip cells like cystidia and basidia are not likely to lie flat therefore making it next to impossible to take accurate measurements!
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gsharpnolack
Registered: 09/05/11
Posts: 657
Loc: Washington State US
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Re: Stropharia ambigua cheilocystidia [Re: inski]
#15797969 - 02/12/12 01:28 PM (3 months, 15 days ago) |
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Quote:
If you don't know how to focus stack I could do it for you on some images if you would like.
Nah, I'm good. But if you find a super pro-intuitive app or site that does it with two or three clicks - I'd appreciate a recommendation.
Quote:
if for no other reason than to protect your objectives from accidental contact with your sample. A cover slip should not change the appearance of microscopic structures.
I haven't had much of a transition into using cover slips. They rarely provide me with anywhere near the same quality of vision as without. Maybe you could provide more specific steps to avoid common mistakes.
I also wanted to ask you if you or anyone else has harmed an objective through contact of any type.
Quote:
with no cover slip cells like cystidia and basidia are not likely to lie flat therefore making it next to impossible to take accurate measurements
I'll keep this in mind
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One thing that can improve our planet: Personally creating a small amount of special atmosphere within the home place.
http://en.wikipedia.org/wiki/Four_Noble_Truths
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Byrain
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Registered: 01/07/10
Posts: 1,819
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Re: Stropharia ambigua cheilocystidia [Re: gsharpnolack]
#15798038 - 02/12/12 01:40 PM (3 months, 15 days ago) |
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Quote:
gsharpnolack said:
I haven't had much of a transition into using cover slips. They rarely provide me with anywhere near the same quality of vision as without. Maybe you could provide more specific steps to avoid common mistakes.
Thinner/smaller sections, the thinner the better.
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 Alan Rockefeller
Mycologist
Registered: 03/10/07
Posts: 24,722
Last seen: 4 days, 21 hours
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Re: Stropharia ambigua cheilocystidia [Re: inski]
#15798133 - 02/12/12 01:57 PM (3 months, 15 days ago) |
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Quote:
inski said:
Agreed, not only for the reasons stated above, with no cover slip cells like cystidia and basidia are not likely to lie flat therefore making it next to impossible to take accurate measurements!
I think dry mounts are useful for making art but not so much for science or mushroom ID.
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gsharpnolack
Registered: 09/05/11
Posts: 657
Loc: Washington State US
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Here's an attempt to rehydrate a gill and add a cover slip...
A slightly dehydrated gill is placed on a slide
A single drop of koh is added
Within a couple of minutes the gill is rehdrated
The excess koh - at least a lot of it - is soaked up using the edging of a napkin
The same cardboard that originally encased a razor blade is used to hold the gill in place as a preparate cut-out is made
The cutout will eventually reveal the cheilocystidia on the true gill edge, while the pleurocystidia (if present) will show itself on either of the left or right false edges
A cover slip is added - perhaps without enough removal of the liquid koh
Dilemma: The cover slip combined with the koh creates a border area blocking the view of cheilocystidia and all protruding features
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One thing that can improve our planet: Personally creating a small amount of special atmosphere within the home place.
http://en.wikipedia.org/wiki/Four_Noble_Truths
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TimmiT
Registered: 03/23/10
Posts: 2,431
Loc: Victoria
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Re: Stropharia ambigua cheilocystidia [Re: gsharpnolack]
#15799658 - 02/12/12 06:39 PM (3 months, 14 days ago) |
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You should have more liquid than that under your cover slip. There shouldn't be any air under it (the last photo shows an air bubble against the edge)
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"Reality leaves a lot to the imagination" ~ John Lennon
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Alan Rockefeller
Mycologist
Registered: 03/10/07
Posts: 24,722
Last seen: 4 days, 21 hours
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Re: Stropharia ambigua cheilocystidia [Re: TimmiT]
#15799679 - 02/12/12 06:45 PM (3 months, 14 days ago) |
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To add water, you can put a drop of water next to the cover slip right on the edge and it will get sucked under the cover slip.
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TimmiT
Registered: 03/23/10
Posts: 2,431
Loc: Victoria
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Re: Stropharia ambigua cheilocystidia [Re: gsharpnolack]
#15799834 - 02/12/12 07:18 PM (3 months, 14 days ago) |
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I'm bored so I decided to make a cover slip pictorial. It's not really complicated enough to need one but what the hell. I mounted a piece of leaf in red wine just to show the process and make things easy to see in a photo. There is otherwise no point mounting a leaf in red wine.
1. Leaf on slide
2. Add a drop or two of your mounting medium
3. Add cover slip. Apply at an angle to reduce bubbles.
4. The outcome
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"Reality leaves a lot to the imagination" ~ John Lennon
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TimmiT
Registered: 03/23/10
Posts: 2,431
Loc: Victoria
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Re: Stropharia ambigua cheilocystidia [Re: TimmiT]
#15799854 - 02/12/12 07:22 PM (3 months, 14 days ago) |
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Quote:
I also wanted to ask you if you or anyone else has harmed an objective through contact of any type.
You could easily damage the objective but the most likely outcome is that your objective will get very dirty and/or stained., which is going to degrade the image significantly.
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"Reality leaves a lot to the imagination" ~ John Lennon
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gsharpnolack
Registered: 09/05/11
Posts: 657
Loc: Washington State US
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Re: Stropharia ambigua cheilocystidia [Re: TimmiT]
#15799975 - 02/12/12 07:43 PM (3 months, 14 days ago) |
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Both comments were spot on.
For others reading, I was able to use the identical slide and specimen. I used a razor blade to slowly lift the cover slip and add about four drops of condo red to provide a light pool of liquid for the entire cover slip dimension. I was able to see a few cheilocystidia.
I then prepared a second slide with a new section, this time rehydrating with a drop of koh, soaking up the excess, adding a cover slip, then adding a few drops of 99.5% water / 0.5% soap at the edge of the cover slip. The water was gradually exhumed under the slide. No congo red was used and I was able, in this instance, to see even better.
Thank you
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One thing that can improve our planet: Personally creating a small amount of special atmosphere within the home place.
http://en.wikipedia.org/wiki/Four_Noble_Truths
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Gravija
Registered: 06/28/11
Posts: 1,766
Loc: Chicago, Illinois, USA
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Re: Stropharia ambigua cheilocystidia [Re: gsharpnolack]
#15800008 - 02/12/12 07:52 PM (3 months, 14 days ago) |
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Everyone who has contributed to this thread is a hero.
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Alan Rockefeller
Mycologist
Registered: 03/10/07
Posts: 24,722
Last seen: 4 days, 21 hours
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Re: Stropharia ambigua cheilocystidia [Re: gsharpnolack]
#15800960 - 02/12/12 10:40 PM (3 months, 14 days ago) |
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Quote:
gsharpnolack said:
For others reading, I was able to use the identical slide and specimen. I used a razor blade to slowly lift the cover slip and add about four drops of condo red to provide a light pool of liquid for the entire cover slip dimension. I was able to see a few cheilocystidia.
The way I was taught to stain things is to mount in KOH or water, then put a drop of stain on one side of the cover slip and a paper towel to wick away the water on the opposite side. The stain goes under the slide and into the sample. Once it is thoroughly red, you put drops of water on one side and keep moving the paper towel on the other side to a dry spot so it pulls the water under the slide and the excess red clears.
Thats not how I do it though, I put the stain on the sample, and if I want it really stained a lot I heat it from below with a lighter until its hot. Then I put the cover slip on, put a paper towel on one side and drops of water on the other until all of the excess stain is gone and the background is clear.
Quote:
They are generally star shaped or stellate with many pointed projections and are very interesting as it has been shown that they are able to trap and kill nematodes, the nematodes are consumed by the fungus and it is speculated that these fungi are obtaining nitrogen from the trapped nematodes to aid in survival.
Here is another paper on fungi eating nematodes. Ability of wood-decay fungi to prey on the pinewood nematode, Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle.
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