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Invisiblecheesenoonions
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Question about Microscopes and IDing
    #1041979 - 11/10/02 10:10 PM (21 years, 8 months ago)

Can someone tell me what kind of magnification I would need to see some microscopic features of various species. I have read that there ae differences in the gills that can distinguish between similar species. Do you need an electron microscope or would a 100x or 50X objective get the job done?

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OfflineRoKiSdEaD
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Re: Question about Microscopes and IDing [Re: cheesenoonions]
    #1042209 - 11/11/02 12:14 AM (21 years, 8 months ago)

I too have been thinking about getting a microscope, but wonder what kind to get. They had one at walmart, up 1000x magnification for 49 bux. Would that work good? I dont know shit about microscopes, and don't want anything fancy, just something that will work to look at spores with. And it must be cheap :smile:

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Invisibleangryshroom
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Re: Question about Microscopes and IDing [Re: cheesenoonions]
    #1042456 - 11/11/02 02:40 AM (21 years, 8 months ago)

You can use a magnifying glass to look at gills. :wink:

If you want to look at spores, and be able to ID from them, which is pretty damn positive as you can get with the experience, you would need something more than a 400X. Ive looked at them under that magnification, and they are still pretty hard to make out.

1000x would be great.

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OfflineToxicManM
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Re: Question about Microscopes and IDing [Re: cheesenoonions]
    #1042928 - 11/11/02 09:15 AM (21 years, 8 months ago)

For looking at spores 400x is probably the minimum. If the microscope has more magnification than that it needs to be an oil immersion objective. If you don't already know how to use one, get a book on microscopy or take a class. For general looking at things I generally use about 400x. When you want to measure spores or look at surface ornamentation you really need oil immersion.

I haven't seen the WalMart $49 microscope, but if it looks like a cheap toy then it probably is. Check around on the internet and at local scientific supply companies. A good student microscope can be had for a couple hundred bucks.

Once you have the microscope, you should also get a book on mycology with one. I recommend 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. It's inexpensive and tells you everything you need to know about the mycology side of things.

Happy mushrooming!


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Anonymous

Re: Question about Microscopes and IDing [Re: ToxicMan]
    #1050297 - 11/13/02 07:30 PM (21 years, 8 months ago)

i've been considering buying a microscope for about a half a year now... there are many high and low end microscopes available on ebay... this is one good example... it comes from a very reliable ebay seller... 1600X... oil immersion and has photographic/video capability...

the current auction thread for this model is http://cgi.ebay.com/ws/eBayISAPI.dll?ViewItem&item=733399648 .... a local dealer would sell a similar "used" scope for about $500 more...


there are many others models available from this and other sellers... some for as low as $100...

i hope posting this ebay link doesn't cause problems... if it does let me know and i'll edit this... i hope this helps u... peace 

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OfflineToxicManM
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Re: Question about Microscopes and IDing [Re: Anonymous]
    #1050742 - 11/13/02 09:47 PM (21 years, 8 months ago)

That particular one looks pretty good. It's better than the one I have.


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OfflineRoKiSdEaD
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Re: Question about Microscopes and IDing [Re: ToxicMan]
    #1050787 - 11/13/02 09:58 PM (21 years, 8 months ago)

the one at walmart is kind of plastic looking... so it must suck,eh? What is oil immersion anyway? I mean, would that just be putting the spores in oil or what? heh, thats what it sounds like (sorry if i sound dumb)

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Offlinezeronio
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Re: Question about Microscopes and IDing [Re: RoKiSdEaD]
    #1051321 - 11/14/02 12:35 AM (21 years, 8 months ago)

Oil immersion is needed for higher magnifications. You immerse the objective of the microscope in a drop of oil so there is no air between the cover glass and the objective.

Edited by zeronio (11/14/02 12:42 AM)

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Invisibleangryshroom
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Re: Question about Microscopes and IDing [Re: zeronio]
    #1051367 - 11/14/02 12:49 AM (21 years, 8 months ago)

yup yup.


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Invisiblecheesenoonions
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Re: Question about Microscopes and IDing [Re: ToxicMan]
    #1053953 - 11/14/02 08:52 PM (21 years, 7 months ago)

What will 100x oil immersion let you see?

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InvisibleCow Shit Collector
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Re: Question about Microscopes and IDing [Re: cheesenoonions]
    #1054270 - 11/14/02 11:20 PM (21 years, 7 months ago)

isnt this also called priming the slide?


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InvisibleZen Peddler
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Re: Question about Microscopes and IDing [Re: Cow Shit Collector]
    #1054647 - 11/15/02 02:44 AM (21 years, 7 months ago)

On the gill surface there are basidia that have spores attached to these cells - often 2 spored or 4 spored. There are also sterile cells called pluerocystidia and cl. cystidia and these can vary greatly in shape - these are often used by the likes of Guzman and other acclaimed mycologists


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Offlinezeronio
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Re: Question about Microscopes and IDing [Re: cheesenoonions]
    #1054723 - 11/15/02 03:33 AM (21 years, 7 months ago)

With 100x oil immersion objective and 10x eyepiece you get 1000x magnification.
Maybe you could se spore ornamentation, clamp connections, count the number of kernels in cells, ...

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OfflineToxicManM
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Re: Question about Microscopes and IDing [Re: cheesenoonions]
    #1055138 - 11/15/02 08:49 AM (21 years, 7 months ago)

zeronio pretty much has it. Greater magnification lets you see smaller and more subtle details of what you're looking at. For example, there are spores which are ornamented, but where the ornamentation is subtle enough that you won't see it without the use of an oil immersion objective (Panaeolus foenisecii is an example of this).

Don't forget to use the immersion oil both between the slide and objective and between the condenser and slide. The oil allows the light cone to be wider (without going into detail, it has to do with refractive indexes) which is necessary for the optics to perform as designed.


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OfflineAnnoA
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Re: Question about Microscopes and IDing [Re: cheesenoonions]
    #1055623 - 11/15/02 12:11 PM (21 years, 7 months ago)

>What will 100x oil immersion let you see?

Something like this, in terms of spores, 1000x oil immersion.

http://www.fungifun.org/gallery2/view_album.php?set_albumName=microscopy
http://www.fungifun.org/gallery2/view_album.php?set_albumName=wood_lovers

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OfflineRoKiSdEaD
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Re: Question about Microscopes and IDing [Re: Anno]
    #1056980 - 11/15/02 09:42 PM (21 years, 7 months ago)

Man this crap sounds too hard to me :frown: I was hoping to stick shit under a microscope and get an ID.... but now i have to do all kinds of crazy things.... i dunno if ill get one or not, im awfully lazy :smile: Well i will, but i dont know how soon, especially since 49.97 aint gonna cut it :frown:

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Anonymous

Re: Question about Microscopes and IDing [Re: ToxicMan]
    #1057772 - 11/16/02 07:42 AM (21 years, 7 months ago)

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OfflineToxicManM
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Re: Question about Microscopes and IDing [Re: ]
    #1057933 - 11/16/02 10:39 AM (21 years, 7 months ago)

Sure.

Similar to the way a drop of oil is placed on top of the slide and the objective is rotated into the oil, a drop should be placed on top of the condenser and the condenser raised until the oil is in contact with the slide.

The details of why can be found here (it's mentioned in the sections following Figures 3 and 6). I suppose I should mention that this needs to be done only if your condenser has a numerical aperture greater than 1.0. Of course, if your oil immersion objective has an aperture of 1.25 (for example, that's what my 100x is) and your condenser an aperture of 1.0, in reality the condenser limits your objective to 1.0 whether you use the oil or not. And failing to use oil on the condenser cuts its aperture to 1.0 (if it was greater than that), which limits the entire optical system to 1.0.

If you don't need the full resolution of the optics and just want the magnification, leaving out the oil on the condenser will save a little time. But if you're going to do photomicroscopy or really detailed work then you shouldn't leave this part out.

Hope that's helpful!


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Invisiblecheesenoonions
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Re: Question about Microscopes and IDing [Re: ToxicMan]
    #1058100 - 11/16/02 12:45 PM (21 years, 7 months ago)

ok, if you want to see how many spores are on a basidia, how would you go about making the slide? I have read that the number of spores attached to the basidia can distinguish species from one another.

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OfflineToxicManM
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Re: Question about Microscopes and IDing [Re: cheesenoonions]
    #1058399 - 11/16/02 03:46 PM (21 years, 7 months ago)

Use a tweezers (or something similar) to break off a couple millimeters of a gill near the stem. Put it on a slide, and add a drop of water. Particularly if the gill and spores are pale you might use a stain (with or instead of water). Put a cover slip on it. Use a pencil (eraser end) to press on the cover slip to sqeeze out air droplets and squish the gill piece a little, spreading out the tissues a bit. Examine under the microscope. Look along the edges of the gill piece. You can generally find a few basidia sticking out where you can see how many spores there are. Note also that you'll usually have to move the focus up and down to count the spores on even a single basidium, due to the low depth of focus of most microscopes.

There are more involved ways (cross sectioning gills) which will give prettier results, but this way is quick and easy.

You're correct that you can identify a few species of mushroom this way, but there aren't that many. Agaricus bisporus is the most obvious example. The vast majority of mushrooms you'll find will have four spores per basidium. Most of the time you'll need to measure the size of the spores, look at shape and ornamentation, and see how they stain (particularly with Meltzer's reagent).


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Anonymous

Re: Question about Microscopes and IDing [Re: ToxicMan]
    #1058500 - 11/16/02 04:30 PM (21 years, 7 months ago)

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Re: Question about Microscopes and IDing [Re: ToxicMan]
    #1059026 - 11/16/02 09:47 PM (21 years, 7 months ago)

Toxicman great info! Would you like to try your hand at some aussie natives that i have here??


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Invisiblecheesenoonions
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Re: Question about Microscopes and IDing [Re: ToxicMan]
    #1060518 - 11/17/02 05:45 PM (21 years, 7 months ago)

Yes indeed!  You have provided e with all I need to know. Thank you!  I may be wrong, but the two species i was thinking of discerning was Pan semiovatus and antillarium.  I read somewhere that they have different numbers of spores per basidia.  Maybe I'm going about it the wrong way.  Any thoughts?  Ohh man my 200th post  :laugh: 

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OfflineToxicManM
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Re: Question about Microscopes and IDing [Re: ]
    #1060783 - 11/17/02 07:47 PM (21 years, 7 months ago)

MrMushrooms

You're very welcome. I was fortunate to have a very knowledgable teacher who required us to learn these things when I was taught to use a microscope. Thank you Mr. Lemieux.

bluemeanie

Thank you for your kind offer. I currently have no use for spores other than to make slides for future reference (I assume that's what you mean).

cheesenoonions

Panaeolus semiovatus is fairly common here in Colorado, and there are no species very similar to it to confuse things (you never need a microscope for ID for them). If Panaeolus antillarum actually has other than 4 spores per basidium then this type of examination would show you that. I don't have any references to the species which tell me anything about its basidia. In general, if a species has other than 4 spores per basidium then that is pretty useful for identification. The only time I've used it personally was identifying some Agaricus bisporus which somebody brought in to the mycological society meeting.


Happy mushrooming, all!


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