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Microscopic Spore Identification


Rick

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Hello,

I'd like to raise my mushroom interest to the next level and purchase a nice microscope for spore identification. I believe this would help in my mushroom identification along with the aid of photo comparison. There is a firm in Florida, Florida Mycology Research Center, which is willing to sell you identified spore prints at a substantial price. There also is an excellent article in MushroomExpert.com titled "Using a Microscope: Viewing and Measuring Spores". This gives all the great information one should record when identifying spores. It even provides some “through the microscope” examples to show how to measure, identify color and shape, etc. A very nice article, but limited in spore content.

The crux of the problem is now that you have purchased your microscope, gathered your mushrooms, made your spore prints, and placed some spores on a glass slide under the microscope, what do you compare them to??? Thus the problem, there seems to be no reference library available for comparison.

For those of us who are not 100% positive on our find, certainly can compare our find to pictures in a book or online. We probably can even find some information as to the color of the spore print, but as to the spore characteristics themselves, nothing.

So if anyone, expert or novice, can point me to a website that contains a library of spore information, I'd be eternally grateful! This perplexing problem has me holding off on making my jump to the next level and purchasing a fine microscope. Really, with the nice microscopes today that allow photo capture and computer download, it shouldn’t be a problem to capture the spore information needed for a identification library.

Thank you for your consideration to my inquiry.

Rick

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The most detailed and comprehensive spore info I've seen are from Mushroomexperts.com. Dr. Kuo has spore characteristics and microscope pictures on most of the species on his site. (E.G. http://www.mushroomexpert.com/russula_virescens.html).

However, you do have to start with the visual identification of the fungi, then go in to the individual pages to confirm/negate your identification with spore info. I have not yet seen a site which uses fungi spores as the starting keys to identification.

Hope that helps!

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Sorry, if I mislead you as to my order of, or only method of identification! I was thinking on using the more definitive spore print followed by spore identification in conjunction along with the photographic physical attributes. I am not that confident in my identification to just holding up a mushroom and declaring its name, especially if it’s one I intend on eating. I was thinking that the spore is the individual "fingerprint" of the mushroom giving a higher consistent degree of positive identification. Let us not also forget the use of reagents and stains as another tool in our tool bag to use for verification. In short, if the National Audubon Society Field Guide to Mushrooms North America would have gone one step further and included a microscopic photo of the spore (stained or unstained) to go along with the spore description it already gives for each mushroom described in its book, that would be an invaluable source of reference indeed.

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I've been using a scope for about 7 years now. It's not a very good scope (looking forward to an upgrade), but I still get some occasional good information by using it. The combination of spore print color and spore shape is sometimes enough info to seriously consider a genus for ID. Sometimes size of the spores, or the quoteint length/width is helpful. Some genera of fungi feature mushrooms that produce distinctively-shaped spores, like Entoloma. But more subtle spore features are often required to get closer to a species ID. For instance, spore ornamentation. One needs a good 1000x scope in order to see this type of stuff. (My 400x scope will occasionally allow me to distinguish between smooth and roughened/warty spores.) But even this level of information may still be insufficient to get a highly-confident ID. Other micro-characters that are used are: presence of types of cystidia, shape/size of cystidia, cellular structures in various parts of the mushroom.

One online resource where micro-characters of mushrooms are often discussed is Mushroom Observer. Here's one of my recent posts where I included a few photos snapped through the lens of my scope. Note that nobody who has viewed this post has offered a solid suggestion as to the ID. The spore size, however, was sufficient for me to rule out one species name that had been proposed.

http://mushroomobserver.org/117308?q=tMbd

Like Planets says, Mushroom Expert has a lot of good micro info.

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Hello Dave-

Thank you for the reply. It is great to hear that someone does actually use a microscope to aid in the identification of mushrooms. I have to agree that the 400x is probably a little light in the power department. From everything I've read, 1000x seems to be the ticket. The Mushroom Expert does have some nice photos of spores doesn't it? The amyloid warts which seem to appear on Lactarius deceptivus would certainly be a telltale sign of identification I'd think.

The color changes with KOH would be of value also, and let us not forget how nice it would be to have some Melzer's solution in our tool kit. I just don't have the connections to obtaining it and I do work in a hospital!

I think I'm going to put my microscope up towards the top of my "wish and want" list and see if I can't start my own little library of spores. I think you've convinced me that it is a worthwhile endeavor.

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Rick, a friend of mine managed to convince his medical doctor to write him a prescription for Meltzer's. I'm down to my last couple of drops, so I'm gonna try this next time I visit the Doc. According to Rod Tulloss (Amanita expert) the prescription method is completely legitimate.

KOH (4-5%) and Congo red are two liquid ingredients that are helpful when using a scope. I mount my spore samples in KOH and I use Congo red to try to flesh out structural characters like cystidia. With a 1000x oil-immersion scope things change. I have yet to use one.

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Dave,

I found this website, http://www.first-nature.com/fungi/index2com.php, in my search for spore information. It is the most extensive to date that I've found. I'm sure you'll find it useful. I'll have to try asking my doc for the script, although with todays legal issues, I might not get it. Although if I explain to him that should I eat an Amanita thinking it to be a Horse mushroom after asking for the solution, the onus might be on him! It'll be interesting to see what he says.

Rick

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Nice website, Rick. I have bookmarked it.

But I did notice a few reports of spore-print colors that are likely incorrect. the site lists several Psathyrella species as having black prints. The only Psath I know which has a truly black print is P. velutina... which has now been placed into a new genus Lacrymaria. Some Psaths have very dark brown to purple-brown prints, but not truly black. IMO, the confusion between dark brown and black results when one wishes to make this determination based upon a print that is taken on a white background. very dark brown looks like black when contrasted with white. A truly black print disappears against a black background, which is the criterion I use to determine "black."

For comparison of spore print colors I use the Online Auction Color Chart. I got it several years back; it cost me $7. This chart is like an extensive color-sample index you may see in a store that sells paint. Colors are numerically coded.

A spore print should be fairly thick before one attempts to discuss its color.

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Dave, All good points on color comparison. I'll have to look into a Auction Color Chart and see if I can pick one up. Glad you liked the website I found. It's a starting point for us that want to go to the next step. As for microscopes, I've kind of narrowed it down to a Meiji MT5310 biological laboratory phase contrast microscope. Well its really a big wish that will take a lot of convincing to the wife! Since I'm still working and 5 years away from retirement, I thought now or never, as after retirement we'll have to survive on cat food and mushrooms! As for another convincing point to the wife, we need it now as we still have a good set of legs under us for all the hiking we'll be doing while we still can. I'm trying hard to cover all the angles!

Dave, you mentioned that you have been utilizing a microscope for a number of years now. Any chance you could provide me a shopping list of the stains/reagents needed for mycology work? I can start searching out the small stuff in the meantime. Any recommendations on slides and slips would also be appreciated. I think I read where some of the slides have a concave center for specimen and fluid retention.

Regards,

Rick

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The slides and slips I've been using for several years now were provided by the Bio department at the college where I work. I just keep washing all this stuff and using again. As for mounting/staing liquids, I use 4-5% KOH for most of my spore mounts. For structural features like cystidia a stain is useful. I use Congo red. I am still awaiting my next doctor's visit, so that I may try to get some Meltzer's. There are other substances that serious mushroom-ID people use. A really good scope is also on my wish list.

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  • 1 month later...

Dave,

I was wondering if when working at the college you ever thought of using a paraffin mount section and a rotary microtome to produce thin sections for viewing?

I wonder if a microtome would work for this rather than trying to get a thin sample by hand with a razor blade. There seems to be a fair number of units for sale on Ebay.

Rick

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My background in microscopy is very basic. I have an old 400x hand-me-down which I got for $50 from the bio department. Over a year ago I was told there would be an inventory of scopes, and that I'd probably have a chance to upgrade to a 1000x which uses oil-immersion. But the inventory has yet to be completed.

So I am not familiar with the methods you mention, Rick. But I'll ask some of the folks in the local bio dept for an explanation. Maybe I can even get somebody to show me examples? But I am at a stage in this hobby where I should try to move to the next level. Thanks for the suggestions.

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  • 2 weeks later...

Rick, I'm wondering if you may suggest some particular micro-characters of fungi that would lend themselves particualrly well to the methods you mention. I think the best way for me to learn about these methods is within the context of my hobby. That is, when the mushrooms start popping around here again, maybe I could arrange to use a scope in one of the labs, and with some good assistance, begin to learn some of these techniques. Also, do you know if any of these methods would be apt to produce anything meaningful if dried materials are used?

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Dave,

Of coarse we have the spores to study from our spore drop. Then if one were to take a thin section of the pileus which included the lamellae (gills) perhaps we could also detect the basidium with spores attached. It certainly is an interesting area to develop. I was thinking that since you seem to have access to a biology department, why not take advantage of the equipment and knowledge on using the equipment. I'm sure the department must have a microtome and if so, you should try it out!

One book that seems to be of great interest in this area Dave is titled How to Identify Mushrooms to Genus III:Microscopic Features by David Largent, David Johnson and Roy Watling. The ISBN is 0-916-422-09-7.

I just don't think one can be exposed to too much knowledge in a lifetime! Besides, now that I have my microscope, my daughter-in-law who is a big horse enthusiast said I have to now learn how to do fecal flotation for equine parasites. She claims to have a old vet who is a real nice guy, and one that she has used for many years, she is going to inquire to see if he would be willing to give a class on the subject. She claims I could be very busy up at the barn where she keeps her horses.

So Dave it only seems a natural combination horse poop and mushrooms!

Back to our main topic, I think it would be nice to explore the whole mushroom from a microscopic viewpoint. Will it allow us to identify a "death angel" from a "honey mushroom", I doubt it. We rely on other easier means of identification for that task.

Without knowing the extent of your involvement with the biology department, I would certainly encourage you to make good use of whatever facilities are open to you.

Shoot, I have a couple of oaks that I'm thinking of cutting down just to inoculate with oyster mushroom spores to see if they'll grow. From my reading, that seems to be something that can be achieved without too much trouble.

One of my next things on the list of "to dos" is to research out a Mycology 101 text book. Dave, the book above that I mentioned to you, although it is a good text, it is probably more at a PhD level. I'd prefer to start at the bottom up rather than from the top down if you know what I mean. Oh and I'm still working on the Melzer's issue too. Always something my friend!

Regards,

Rick

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Rick,

Lugol's solution can also be used to test spores as amyloid or not, and does not require a prescription.

For light colored spores (Amanita, etc.) it is just as good. The function of the chloral hydrate is to bleach

darker spores so you can see the color change.

There has been at least one replacement for chloral hydrate developed: http://visikol.com/information.html

It seems to be pretty pricey at present, but I'm glad someone is working on the problem.

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  • 2 weeks later...

Rick, that's an interesting issue... what constitutes a good "Mycology 101" book? I have plenty of field guides, each of which provides some background on mushroom features. But an introduction to mycology...? I am aware of the Largent "ID to genus" series. I really should have these books. Thanks for the reminder. And, I'll really need to see if I can get some access to scopes --and the people who know how to use them-- here at my college. My educational background in in mathematics. Other than what I have picked up here and there about mushrooms, I have no background in biology. But I do know a few things about spore shape/size and cystidea. I do use a junky old max 400x scope at home.

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