Hi all

Hi all

Postby Simon Robson » Mon Dec 21, 2009 9:59 pm

My name is Simon Robson and I live in Jersey C.I.

Today I have just received my first microscope a Brunel sp-100 Biological trinocular compound.

Unfortunately I do not know anyone in Jersey with a microscope so this is all very new :o

The sp-100 came with something to attach (technical term) my 5D and lots of fun things like slides cover glass and a slide micrometer!

So I think I will be pestering the group with a lot of basic questions.

My main interests are fungi, entomology and biology.

Living in Jersey we do not have any microscope shops, but I cannot thank http://www.brunelmicroscopes.co.uk enough. Simply excellent service.

So if any of you have any advice for a complete beginner please feel free :D

I will be ordering stains ect soon.

It has been a horrendous year for fungi in Jersey this year terrible weather, still I have a few ids for the group.

Regards,

Simon
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Re: Hi all

Postby admin » Tue Dec 22, 2009 11:11 am

Hello Simon

It's probably too broad a a subject to deal with comprehensively on the forum. However the ABFG has produced a series of articles in the Forayer covering all aspects of basic fungal microscopy and I am sure we can post off some copies to you. In the meantime other forum members may want to offer their own bits of advice. In any event good luck with the microscopy. You will find it quite rewarding once you get the hang of things.

One thing I would say is that the stage micrometer, if it is the usual one that Brunel send out, is unlikely to be of much use. Its calibrations are not fine enough for calibrating your eyepiece graticule (the measuring line up in the top end of the microscope). But we can put you on to an ABFG Jersey member that I suspect will have a good quality stage micrometer. Contact me directly: mj@abfg.org

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Re: Hi all

Postby jonathan need » Tue Dec 29, 2009 1:47 pm

Hi Simon

Welcome to the group, its good to hear i am not the only one with a new microscope and no idea how to use it properly. :D
Maybe we can help each other along the way.

Jon
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Re: Hi all

Postby Simon Robson » Sat Jan 02, 2010 5:04 pm

Hi Jonathan, Thanks for the welcome.

As for the microscope I can only assume that I died over x mass and am now in purgatory.

I did not know the first thing about them (apart for needing one) but I must confess the so and so has on more than one occasion nearly gone through the window.

I know it’s me and my incompetence”this makes it worse”

First was I had no idea you would need another light (from above)
Depth of field is worse than my 2.8 macro
The 100 thing (with the oil?) hits the slide before focusing
As for the thing to attach my 20d too don’t go there

As if Fungi and Lichens where not hard enough …LoL


I know I will need some reagents ect so I will order some now the holidays are over.

Hope you’re having more luck than me.


Best Wishes

Simon R
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Re: Hi all

Postby admin » Sat Jan 02, 2010 8:55 pm

Simon, Happy New Year (tears and all). But something is clearly wrong here. The Brunel Winchester SP100 Kohler has built-in illumination and has no need of lighting from above. Depth of field is always very limited with transmitted light through any compound microscope, but that won't stop you getting needle sharp images and it's why you have fine focusing adjustment. The high power oil immersion is only swiveled into position after you have focused accurately through x10 and x60. You wouldn't swing it into position without oil, and without first getting the focus right with the low power objectives. The only reason it would hit the cover slip would be if the cover slip was extraordinarily thick, or the instrument is faulty (unlikely with Brunel)

Give me a call if you want to - the number is in the back of the Forayer. We need to sort you out quickly, or it will feel like purgatory!

MJ
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Re: Hi all

Postby admin » Sat Jan 02, 2010 8:58 pm

Simon

A basic afterthought. Are you by any chance trying to use this instrument to view the surface of a solid object?

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Re: Hi all

Postby admin » Sun Jan 03, 2010 10:36 am

Simon

Sorry to post this reply in separate bits, but the more I read into your original post the more I have come to the conclusion, perhaps wrongly, that you are attempting to use the microscope in a way for which it is not designed. There are basically 2 kinds of light microscope: a low power stereo dissecting microscope, which you use with reflected incident light from above to view a solid object e.g. the surface of a feather or a leaf or a piece of fungus; and a high power compound microscope, which works with transmitted light that passes through a very small amount of material from below. Your SP100 is designed for the latter purpose, which is why it has no light source from above and no depth of field and, I suspect, why the x100 is hitting the cover glass. You can make a squash of a piece of material about the size of a pinhead, using a macerating fluid like KOH where necessary, or use a razor blade to cut a very, very thin slice that is not more than about 1/10th of a millimetre thick and therefore effectively transparent, or make something like a spore print. But the space between the glass slide and the cover glass has to be so fine (a fraction of a millimetre) that a single small drop of water will fill the entire space.

Now, am I right or barking up the wrong tree?

MJ
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Re: Hi all

Postby Simon Robson » Sun Jan 03, 2010 11:33 am

Hi Michael thanks for taking so much time.

You are quite correct in thinking operator error, On a major long stint with the beast this morning everything in your last couple post is correct.

What a Muppet I am.. LoL :oops:

I vastly underestimated the size of the sample you could use, I need a microscope to cut the bits small enough.

Its still is going to be a labour of love but a least thanks to you I hope I am on the right track.

I do not have any stains or KOH at the moment would NaOH (caustic soda) diluted 10% do the job?


Thanks again for your time


Best wishes,

Simon Robson
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Re: Hi all

Postby admin » Sun Jan 03, 2010 11:56 am

Simon

No worries! We all have to start somewhere, and good for you to take up the challenge. Caustic soda diluted might work temporarily but the problem may be contamination. Reagents for microscopy need to be very pure or what you see down the tube looks like a rubbish dump!

The best way to view bits of fungal tissue when you are just starting out, is to take a specimen and using either a razor blade or a scalpel cut a very small piece (about 2-3mm square) out of the edge of a gill. Pick it up with a needle (dampened) and lay it flat on the slide. If it is a tiny delicate gill, as in a Mycena, just add a single drop of water and lay the coverslip gently on top. If it's a square coverslip, lower one edge onto the slide, place a needle under the slip to support the open side and gently lower it like a hinged lid, watching as the water drop spreads underneath, that you don't trap air bubbles. If it's a thicker gill, apply a drop of your Caustic Soda, which will start to macerate the material. Always start with the x10 objective (or whatever low power there is on the instrument) and then work up through the magnifications, refocusing each time. For the oil immersion objective you can temporarily use pure olive oil or sunflower oil - a single small drop and be sure to wipe the lens afterward with a soft lens tissue. The oil forms a 'bridge' between the surface of the coverslip and the surface of the lens, which has a better refractive index than air and means you don't see a fuzzy image.

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Re: Hi all

Postby Simon Robson » Sun Jan 03, 2010 12:20 pm

Thanks again for the advice, although my ignorance knows no bounds.

I did not know what the immersion oil was for most had leaked to in transit but there is still some left. Did not occur to me to put it on the coverslip…

Best Wishes


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