Request for Microscopy Techniques/Info

[Lancashire Lad]

OK fungifans, after much “shall I or shan’t I” soul searching, I’ve finally come over to the dark side, and bought a microscope.

I’ve spent a couple of days getting used to the basics of setting it up, - (don’t have any immersion oil yet, so haven’t sorted the proper use of the 100x lens), calibrated with stage micrometer & eyepiece graticule, and am generally seeing some nice clear views of whatever it might be that I’m looking at.

And therein lies the route of many problems – trying to decipher what it actually is that I’m looking at, and worse, how best to prepare the specific microscopic parts of the specimen when I do know what I’m looking for.

I know what spores and the asci containing spores look like, and how to go about finding them, but what about such things as cystidia, basidia, clamp connections etc. etc? – I understand that there are several types of cystidia, and that they appear on different parts of the fungus.

I acknowledge the information in the “Missing Basidia” thread, but can anyone give any further information on the best ways of preparing the different types of cystidia etc. for microscope viewing? – i.e. where to take the samples from and how best to prepare them on the slide. – Should they always be looked at dry initially, or wet? With or without coverslips? – Do you use plain water, or some other solution? (Is there any good source for such information? – I haven’t found much about this on the web).

Is there a general “checklist” or “order” of things that should be looked for with a typical fungus? – (bear with me ). i.e. something like: - first look for spore colour, second size the spores, third look for ornamentation, fourth look for cystidia, fifth look for clamp connections etc. etc. – In what order do you experienced microscopists go about things?

Any pointers will be much appreciated. I will try to upload photos of things as and when found, to accompany any future queries – of which there are bound to be many!!

Just as an afterthought, would it be better to start new threads for any such advice? With separate titles such as “How to prepare cystidia” etc.? – That way, any detailed advice will remain searchable, as per Chris’s “Stains in Microscopy” thread, and wouldn’t get lost as individual posts within threads might.

Regards,
Mike.

[Chris Yeates]

Clearly you're asking for lots of details, Mike

this is a first response with just a few first (quick) thoughts; fungal microscopy is a shallow learning curve (in the true sense of that much misunderstood phrase!); you have to take it slowly.

Basically -

• well done! it'll be frustrating at times (it isn't an immediate identification solver, believe me) but you'll not regret it - when I started out there was nothing like WAB for example . . .
  
• don't bother about oil at this stage
  
• I'd recommend getting a x60 objective if you haven't already got one (one of the best bits of advice I was ever given)
  
• practise with a shop-bought ripe mushroom - using a scalpel or similar try removing a gill, lay it flat on a slide - no water, no stain at this stage - and using a low power objective (try the x10 for starters) focus down towards the gill surface - if you can see groups of four (like the dots on dice) they've sold you the wrong thing - rather rarely among agarics cultivated mushrooms should show a patter of twos; when next you find a wild toadstool, try the same tecnique and you'll almost certainly see those groups of four
  
• don't limit yourself to toadstools - there's lots of other fungi which will often give you better (or at least quicker) results - I'll post more about this in due course
  
• practise getting small amounts of material in your slide preparations - less is almost invariably more
  
• I'm a bit tied up for a month or so - but if at some stage you want to pop over the Pennines, I'd be happy to give you some tips
  
• finally make sure you learn to set the microscope up correctly - so many people don't, and that can be why they get disappointing results
  
• and finally, finally have a look at the have a dung day thread . . . . collect some rabbit pellets and keep them damp as described there - that way you will always have material of lots of different groups to look at with a minimum of effort (dry weather such as we've been having recently needn't stop your progress); it really is a nasty, smelly activity - honest! as long as you stick to herbivores

that should be enough for starters + hopefully there's some useful info on this thread already

cheers from the White Rose County

Chris

[stickman]

Welcome to the dark side Ken

When I first bought my microscope (about a year ago) I was asking much the same questions as you, and was recommended a book by someone (I forget who) on WAB:

'How to Identify Mushrooms to Genus: Microscopic Features v.3' by D.L. Largent (Author), et al.

It really is excellent: It covers all the basic and much of the more advanced aspects of mushroom morphology, stains, technique etc etc

As you would expect from the title, you won't find any info on, say, Ascomycetes but many of the techniques covered are the same whatever you are looking at.

If you can't pick it up for a decent price, I would be more than happy to lend it to you!

Cheers, Nick.

[Lancashire Lad]

Originally Posted by stickman

....'How to Identify Mushrooms to Genus: Microscopic Features v.3' by D.L. Largent (Author), et al....If you can't pick it up for a decent price, I would be more than happy to lend it to you!....

Thanks for the offer Nick, that is most kind of you. I've just done a quick search, and the book, although apparently out of print, seems to be available from several sources, so I'll try to get hold of one.

Regards,
Mike.

[earthdragon64]

The following website might be helpful;

Under the microscope - Scottish Fungi

Also, depending on how far you want to go, and how deep your pockets are, Liz Holden holds a 5 day Identifying Fungi Workshop which has quite a bit of microscopy work in it. It is held at the Field Studies Council centre at Kindrogan;

Workshops - Scottish Fungi

I've not done microscopy with Liz, it's a course I'd like to do in the future when I've got time though. I have been on some forays and day long ID courses with Liz and would highly recommend her as a teacher, her knowledge and enthusiasm seem boundless.

Regards, Audrey.

[Lancashire Lad]

Chris,

Thanks for this reply. You have provided some good initial pointers here.

I've got x4, x10, x20, x40 and x100oil immersion Infinity Corrected Plan Achro objectives, and x10 widefield eyepieces.
Not sure if a x60 is available but will check with them next week.
I know there is a x80, but it's about £160.00, so I wouldn't want to spend that amount extra unless you think it would be a useful and worthwhile compromise between the x40 and the x100immersion lens?

If both x60 and x80 are available, which would you recommend?

I will have a go with shop-bought mushrooms. Off to the shops shortly.

Yes, I've already discovered that what looks small to the eye, still looks like a great boulder when under the scope. So getting the smallest amount of material under the lens seems to be the order of the day!

I've got various information on setting the scope up. - I have centered the condenser properly, and have the theory on adjusting it, but I must admit that I don't seem to notice much change when altering the condenser position & diaphragm aperture.
There definitely is a change - in the appearance of cell walls etc., but not very much.
Perhaps it's just a case of getting used to the extent of the difference or maybe I'm not yet adjusting it correctly?

Originally Posted by Chris Yeates

....it really is a nasty, smelly activity - honest!....

-I hope it isn't!!

This morning, whilst searching for the book which Nick mentioned, I stumbled upon this website page: -
Using a Microscope (MushroomExpert.Com)
which has links to several primer pages giving exactly the sort of information I was looking for on cystidia, basidia, etc. etc.
To me, the information via these links seems to be pretty comprehensive and useful. (But I'd still like to see further information in due course from yourself and from other experienced microscopists. The more info that I and others can get, the better.).

Lastly, thanks very much indeed for your offer of giving me some "hands on" tips. Huddersfield isn't a million miles away, so I may well take you up on that when you've got some free time. I would undoubtedly learn more from yourself in an hour, than I would in a week from the web or books.

Best regards,
Mike.

[Lancashire Lad]

Originally Posted by earthdragon64

The following website might be helpful;

Under the microscope - Scottish Fungi

Also, depending on how far you want to go, and how deep your pockets are....

Thanks very much for this info Audrey.
Just had a quick look, and the Scottish Fungi site looks to be very useful!

As for the depth of my pockets - well they are, unfortunately, pretty shallow. - Even more so just now, having just bought this microscope kit. - If I had any moths in my wallet, they will have definitely now left due to lack of food.

Regards,
Mike.

[Chris Yeates]

Originally Posted by Chris Yeates

[*]and finally, finally have a look at the have a dung day thread . . . . collect some rabbit pellets and keep them damp as described there . . . . . it really is a nasty, smelly activity - honest! as long as you stick to herbivores[/list]Chris

did I say that?

I did of course mean NOT

Chris

[Chris Yeates]

Originally Posted by Lancashire Lad

. . . . .

If both x60 and x80 are available, which would you recommend?
. . . . .

Best regards,
Mike.

see: SP200 Accessories (x60 over x80 every time); just a quick bit of advice - and if this is obvious to you I apologise - if you are tempted by higher power eyepieces (x15 or x20) remember that you are just making an already existing image bigger - not improving resolution (they are not the same thing at all) so they are an unnecessary luxury at this stage, if you were wondering about them

with regard to other stuff make sure you have (at least) a scalpel or two and fine-pointed forceps - don't worry about stains at this stage - if and when you come over I can supply you with starters on those . . .

you're not going to thank me when I say that a dissecting microscope is exceptionally useful as well () - unfortunately another WABber now has my old one . . . but see:

Fungal Plant Parasites

I think the item linked to is no longer available from that site, but also see: QUICKTEST Small magnifiers on stands (including linen provers / linen testers) (I'm thinking particularly of that "10X30 Loupe on a Stand") it means you can do stuff like gill removal, preparing a gill edge on a slide etc. hands-free

cheers

Chris

[Lancashire Lad]

Originally Posted by Chris Yeates

....(x60 over x80 every time)....

Cheers. - I'm not sure if the SP200 accessories will be compatible with my scope, but will check regarding x60 on Monday.

Originally Posted by Chris Yeates

....(just a quick bit of advice - and if this is obvious to you I apologise - if you are tempted by higher power eyepieces (x15 or x20) remember that you are just making an already existing image bigger - not improving resolution (they are not the same thing at all) so they are an unnecessary luxury at this stage, if you were wondering about them)....

No problems - other eyepieces aren't on the agenda, but I fully accept what you are saying.

Originally Posted by Chris Yeates

....with regard to other stuff make sure you have (at least) a scalpel or two and fine-pointed forceps - don't worry about stains at this stage - if and when you come over I can supply you with starters on those...

I have scalpels and very fine pointed forceps. I think I'll need to get some single sided safety razor blades though.

Originally Posted by Chris Yeates

....you're not going to thank me when I say that a dissecting microscope is exceptionally useful...

I have some devious plans in that direction.

In the meantime, I have something very similar to these which I find useful: -
Mains Desktop Illuminated Magnifier - Stores and Prices
It may not be the best, but mine has x2 large lens, and x 5 smaller inset.

I will see how things go but the dissecting scope would obviously be the best solution.

Regards,
Mike.