Tubaria furfuracea - spores

[mebeingme]

Ok, so not really Malcolm Storey’s league, but I guess we all start somewhere…





I’ve finally taken the plunge and experimented with my new microscope which has been sitting unused for some time now. I thought I’d try and look at spores of Tubaria furfuracea as this species is in abundance at mo. I prepared slide by slicing a 2 mm square of gill area (fresh), transferred to slide, added small droplet of water to gill area and sealed using glass cover square. (Re-reading this description - I sound like Professor from Fast Show!) I had difficulty analysing spores this way and had more joy when I cut a smaller piece of gill and looked directly on slide. Saying that, photos were still disappointing. I had been advised to squash a piece of gill tissue the size of a pinhead between a slide and cover slip but had difficulty cutting smaller piece; also am not sure whether water / oil would be advisable to moisten tissue sample? – had a fair few air bubbles using former.

Having left the entire cap on slide overnight, I had an amazing spore print following morning. I have checked fringes of this under microscope @ x 400 magnification and am really pleased with results.
The spores seem to tick all the boxes with Roger Phillips description of this species – “spore print was pale ochre and the spores do appear elliptic with rounded apex. The only down side is that I haven’t yet worked out how to measure spore dimensions: My microscope has a built in LCD screen instead of eyepiece so am not sure whether I can use a calibrated graticule.

Any thoughts always appreciated

[SheffieldLass]

Congratulations!

Spore prints onto slides are the easiest way to see the spores, at least you know they are all mature, and they are not hidden by other bits of the fungus structure.

It looks as if you were looking at the spores dry. It is easier (usually, unless you have only two or three clear ones, which escape when you put water on them ) to look at them with water (or if clear ones, with stain), with a cover slip, and when you do come to measure them, the size is more accurate. They do measure a different size when dry compared to wet, and the sizes you see quoted are ones measured wet (or so I understand).

Sounds as if you tried the right technique with the bit of gill. Small is best though, but can be tricky. When you first mount it, add water or stain, put on a cover slip and squash lightly. You should be able to pick up a bit of the structure, particularly things like cheilocystida, the specialised cells on the gill edge, even if you can't see them clearly yet. You may then need to squash it further to get a clearer picture of the individual cells. The more you squash it, the more the structure of the gill gets mixed up though, so at that stage it may become difficult to work out which are cheilocystidia (on the gill edge) from pleurocystidia (which occur within the gill, not the edge), hence why to do gentle squash first.

Anyway, you've got a whole new world opening up for you now!

As for measuring spores or cells etc with your set up .... if you use a stage micrometer (a slide which has been calibrated), you could capture a frame of that at each magnification level, which would appear as a scale rule in the photos. You then have a reference scale you can compare with. Maybe draw from that a rule on paper which you can then put against the screen for captured pictures of the fungi taken at the same magnification. Hope I've described that so you understand what I'm talking about ....

Have fun

Melanie

[Ditiola]

As you only need the stage micrometer just the once, it is best to borrow one if you can.

In the mean time you can work out an approximate scale using the spores from Lycoperdon pyriforme as they are very consistent in size. They are 3.5 to 4 microns diameter therefore with a scale based on these spores you are likely to be accurate to 0.5 microns.

You can have hours of fun with your microscope. You can just look at everything even if you do not fully understand what you are looking at, after all that’s what the first Mycologist had to do when microscopes were invented.

Peter

[fairplay]

The only thing nobody has picked up on is your being unsure whether to use oil or water ("to moisten tissue sample")
You sound a bit confused here (or is it me ?) Only water or a stain/reagent is used between the slide and the cover slip.
(Immersion) Oil is only used on the cover slip when you want to lower your (usually 100x) oil immersion lens onto and into the tiny drop of oil placed on the cover slip.

I have a feeling on your microscope set-up, you do not yet have an oil immersion lens available.

NEVER use oil with any objective lens other than one specially made for oil immersion. The lens will usually have "oil" embossed on the side.

Neil.

[mebeingme]

[quote=SheffieldLass;593916]Congratulations!

Anyway, you've got a whole new world opening up for you now!

Yes, I think I am officially addicted

Thank you so much for information, especially use of stage micrometer which didn't know existed. The tip about L. pyriforme is also useful to know.

Neil, I was confused about use of oil or water beneath cover slip so thanks for feedback. I was recently re-reading thread entitled 'Microscope help needed' - posted Sept 2008. A member had encountered similar problems with identical microscope set-up as this model doesn't have a 100x oil immersion lens. It would be useful to contact this member to see whether they were able to acquire this lens - is this possible on WAB? Maybe this same member went on to become Malcolm Storey ?

[fairplay]

Hi there,
I know nothing about the equipment you have, but I would imagine if a 100x lens was available the resolution would be so poor that no further detail could be seen - I may be wrong.

To send a PM, just click on the persons name next to the avatar and a list of options should come up and the first option is to send a PM.

Good luck.

Neil.