help with making microscope slides of fungi

[Walloon]

Can anyone offer advice of how to prepare permanent/semi-permanent microscope slides of fungi.

Specifically, I’d like to create a slide showing a section of gill with a row of basidia lining the edge with spores still attached.

After some trial and error with a razor blade I’m getting reasonable hand-sections that nicely show up the spores/basida on fresh specimens.

The problem comes as soon as I apply a drop of slide mountant however. Within minutes the majority of basidia and spores have disappeared!

Some spores are drifting away in the mounting fluid but even when this doesn’t happen I still lose visibility of most.

My working hypothesis (?) is that the mounting fluid is causing the main mass of hyphae making up the gill body to expand slightly over a period minutes. Since the spores start only a couple of microns out from the gill edge, even the slightest ‘bloating’ / expansion of the gill hyphae is enough to ‘swamp’ the basidia and spores under my coverslip.

I’ve tried glycerine, water, Permount and Loctite as a mountant – all with the same result.

It’s time consuming, so I’ve stopped short of the whole ‘dehydrate/wax-embed/section’ routine, but I’ve the kit to do it if someone says it will help.

Any advice appreciated!

Harry

(p.s Note to forum editors: I’m new to the forum – I’m resending this thread as I wasn’t sure my last attempt registered on the site. Feel free to delete one or the other. )

[Ditiola]

Hi Harry and welcome to WAB

I think you have set yourself a very difficult task, and I am not sure that it will be possible to get a permanent mount with spores attached to basidia. Perhaps if you use immature specimens where the spores have not fully mature you might stand a better chance, but to get a whole row of basidia with spores is I think pretty unlikely. Although I expect some other WAB contributors may have some useful tips.

Peter

[fairplay]

I will be attending a week long microscopy course in Ipswich next year where we will be making permanent fungal mounts (or in my case, attempting to !), but it has already been pointed out to me that it serves no real useful purpose as the mycologist always has fresh material (dried or wet) to work with.

But in my case it will enable me to improve my microscopy techniques and increase my knowledge of chemical use.

I'm still pretty dumb when it comes to manipulating things on the computer so am unable to do a link highlighted in blue, but if you are interested, it is a residential course, run by Ernie Ives, at the Belstead House, Ipswich, Suffolk.

I think you should be able to Google Belstead House residential courses and hopefully something should come up.

Neil.

[SheffieldLass]

You could capture a few microphotos instead .... permanent if burned onto cd

[FungiJohn]

Originally Posted by SheffieldLass

You could capture a few microphotos instead .... permanent if burned onto cd

and quite possibly the best future alternative method

John

[flaxton]

This method allows only the information you need (spores cystidia clamps) to be centre stage in the photo. With a permanent slide a particular cystidia can sometimes be difficult to locate.

Mal

[cybershot]

It's amazing the heights to which you can aspire when considering permanent microscopy records!



David

[cybershot]

Originally Posted by cybershot

It's amazing the heights to which you can aspire when considering permanent microscopy records!



David

Apologies for not making it abosolutely clear that this is the work of Canadian mycologists Jacqueline Labrecque and Roland Labbé.

David

[Walloon]

It's been my first posting to WAB and I've been really amazed at the speed and number of responses. Sincere thanks all!

At the risk of seeming ungrateful however - I'm still not clear if it's possible to make the sort of slide I'm picturing (I should add, my interest is purely that of an amateur hobbyist). Do the Kew's of this world really not have a reference slide collection of such things as gill edge structure?

One thing I feel I must take issue with (that I guess may ignite some discussion!). I really can't go along with the suggestion that "slides arn't needed, photo's are enough". By that logic who'd ever need to see a real mushroom again - there are a million mushroom photos out there.

[FungiJohn]

Originally Posted by Walloon

It's been my first posting to WAB and I've been really amazed at the speed and number of responses. Sincere thanks all!

At the risk of seeming ungrateful however - I'm still not clear if it's possible to make the sort of slide I'm picturing (I should add, my interest is purely that of an amateur hobbyist). Do the Kew's of this world really not have a reference slide collection of such things as gill edge structure?

One thing I feel I must take issue with (that I guess may ignite some discussion!). I really can't go along with the suggestion that "slides arn't needed, photo's are enough". By that logic who'd ever need to see a real mushroom again - there are a million mushroom photos out there.

Hi Walloon and welcome to WAB btw

You could use the comparison (loosely) with how we once looked at video tapes as against DVD, Blue ray etc format.

A slide can only really be viewed using a microscope and generally only one person can view it at a time, where as a digital image can be viewed on many platforms and by many.

Kew, like many other institutions will no doubt have slides but this is essentially old technology.

With regards to ‘mushroom photo’s’ these are still required in order to aid identification. You could compare with others but they are not the ones ‘found’!

I have almost 2 million fungi images of around 1000 species, a figure which seems large but many are still unidentified through lack of microscopic data or available information.

Hope this helps

John