Home | Community | Message Board
You are not signed in.
Sign In New Account
Forum Index Search Posts Trusted Vendors Highlights Galleries FAQ User List Chat Store Random Growery


This site includes paid links. Please support our sponsors.

Mushrooms, Mycology and Psychedelics > Mushroom Hunting and Identification
Threaded Mode Threaded  Previous topic Previous  View all topics Index  Next topic Next 

Welcome to the Shroomery Message Board! You are experiencing a small sample of what the site has to offer. Please login or register to post messages and view our exclusive members-only content. You'll gain access to additional forums, file attachments, board customizations, encrypted private messages, and much more!

Shop: Mushroom-Hut Substrate Bags   Myyco.com Isolated Cubensis Liquid Culture For Sale   Sporeworks.EU Spores for European Microscopy   North Spore North Spore Mushroom Grow Kits & Cultivation Supplies

Jump to first unread post Pages: 1 | 2 | 3 |  [ show all ]
Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
Calibrating Microscope
    #11133509 -

So i got this microscope a month ago about. Ive been enjoying just looking at different spores but i really need to be able to measure them. I read up from Micheal Kuos article. So my question is 1. My microscope doesnt seem to have any sort of measuring system in the Eyepiece. Do i have to put one in? How?  Its hard to start from next to no Experience and go straight from a article. I would really appreciate some advice from anyone Experienced in mushroom Microscopy. im lost.


--------------------

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: ehtdaedlufetarg]
    #11133522 -

i believe what you will be needing is called a reticle

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: ehtdaedlufetarg]
    #11133546 -

You need a stage micrometer that shows a "grid" when you look through the eyepiece. You have to calibrate that.


--------------------
Mr. Mushrooms said:
I will confess something that should be quite obvious, CC.  I love mushrooms, i.e. fungi.  I really do.  I am talking about a strong feeling, i.e. emotion, for them.  I think they are beautiful.  I even dream of them.

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: ehtdaedlufetarg]
    #11133648 -

http://www.microscopyu.com/tutorials/java/reticlecalibration/

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: ehtdaedlufetarg]
    #11133694 -

You need a reticle, which is a little piece of plastic that goes into the eyepiece, so you have a grid when you look through it.

Most people buy a 10x eyepiece with a reticle installed for about $35, but you can buy just the little plastic bit, maybe, but I don't know where.

I like the grid style better than the crosshairs style, its much easier to measure the spores that way.

Always measure spores at the highest magnification you can.

I usually keep the reticle in all the time but that gives some people headaches if their eyes can't ignore it.

To calibrate the reticle you need a stage micrometer, which is a special microscope slide with fine divisions.  They cost between $30 and $100 on ebay.  You only need it once.  I got a nice one for $30 on ebay that was supposed to cost $400, it is made from mirrored glass and has tiny holes.  Anyone who wants to borrow it should PM me.

The divisions on the reticle are usually 1 or 10 micrometers, but using the stage micrometer I found that my divisions are actually 8.7 microns, which is a significant difference and a real good thing to know when calculating spore size.  Cactu's reticle is actually 9.4 microns per each 10 measured.

If you are using a microscope to look at mushrooms, you need "How to Identify Mushrooms to Genus part III: Microscopic Features".  If you have a microscope but don't have a copy, PM me and I will fix that problem for you.

Measuring spores by hand introduces a lot of error.  Its better to use the Mr. Mushrooms method, where you hold a camera to the microscope reticle to photograph the spores and grid, then use a photo editor like The Gimp or Photoshop to count the number of pixels wide and long each spore is.  Then measure the grid, calculate the number of pixels per micrometer, and use math to turn the number of pixels into micrometers.  This eliminates lots of sources of error and gives you measurements that are exact to the nearest ten nanometers, more or less.  Probably less but that doesn't really matter, its cool to be able to say "this spore is 9.67 microns long".

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: falcon]
    #11133860 -

This was helpful to me as well. What measurement type of stage micrometer should we get? My maximum view is 500x.

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: World Spirit]
    #11133862 -

Woops should've refreshed the page first
Thanks AR

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: Alan Rockefeller]
    #11134487 -

Alan Rockefeller said:
Measuring spores by hand introduces a lot of error.  Its better to use the Mr. Mushrooms method, where you hold a camera to the microscope reticle to photograph the spores and grid, then use a photo editor like The Gimp or Photoshop to count the number of pixels wide and long each spore is.  Then measure the grid, calculate the number of pixels per micrometer, and use math to turn the number of pixels into micrometers.  This eliminates lots of sources of error and gives you measurements that are exact to the nearest ten nanometers, more or less.  Probably less but that doesn't really matter, its cool to be able to say "this spore is 9.67 microns long".


Hey, that was supposed to be our secret.  :lol:  Seriously though, thanks for the credit.  I've used both methods but prefer mine for accuracy.

Also, after calibrating my way I merely hold a ruler up to the monitor and measure the spores, doing the math.  I can measure 30 spores in less than 5 minutes.  The standard way takes far longer.


--------------------

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: Mr. Mushrooms]
    #11134576 -

Is Ebay a good place to get a microscope? I am guessing no. If not, where?


--------------------
:macrofunction:

Mushroom Hunting and Identification Rules
Puget Sound Mycological Society
MushroomExpert.com
Sample ID Request - Necessary for proper ID's
An Overview on how to find Psilocybe mushrooms by Shockvalue
P. cinctulus (subbalteatus) vs. P. foenesecii
How do I put pictures in my posts?

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: Twiztidsage]
    #11135419 -

I think Alan has a link to a decent microscope offered on eBay.


--------------------

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: Mr. Mushrooms]
    #11137147 -

Wow. Soo helpful. Thanks guys.


--------------------

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: ehtdaedlufetarg]
    #11137583 -

I want to add to this thread for future search inquiries a note or two on the following unrelated material:

(Not listed at www.dictionary.com is the word pleurocystidia)

pleurocystidia
Plural form of pleurocystidium.

Pleurocystidium -
(mycology) A relatively large cell found on the gill face of a basidiomycete


Jump in and add a significant piece of basic knowledge to help upcoming microscopin' fungofamiliacs

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: Twiztidsage]
    #11137762 -

Quote:
Is Ebay a good place to get a microscope? I am guessing no. If not, where?


Yes.  There are many suitable microscopes on ebay for good prices.  There are also lots of crappy microscopes and also really nice microscopes.  Sometimes for low prices.

This one costs $170 and has achromatic objectives:

http://cgi.ebay.com/BINOCULAR-BIOLOGICAL-MICROSCOPE-w-MECHANICAL-STAGE_W0QQitemZ350179455000QQcmdZViewItemQQptZLH_DefaultDomain_0?hash=item5188527018&_trksid=p3286.c0.m14

There are also many decent microscopes on amscope.com.

Quote:
(mycology) A relatively large cell found on the gill face of a basidiomycete


I wrote that. 

Its now the second google hit when you search for pleurocystidia.  I am so excited.  Its going to make me famous for sure.

Everyone that wants pleurocystidia will have to come to me to get them.  When putting a name on a species its really important to have them yet they can be really hard to find. 

Its easy enough to find them on fresh material - you cut the cap in an extremely thin slice perpendicular to the gills so you have a slice that looks like a very thin saw blade.  The pleurocystidia is on the sides of the saw teeth and the cheilocystidia on the tips. 

But when its dried its a lot harder to find the cells on the gill face.  You can't cut the cap like that, it will just break. Cheilocystidia is no problem.  For the pleurocystidia I guess you can rehydrade and fold the gill over.  But that only gives a narrow section of the gill face and usually there are no pleurocystidia there.  Also the gill usually breaks when you try to fold it over which pretty much sucks.

Maybe the key is to make lots of really thin slices of a bit of gill, and hope that some pleurocystidia shows up exactly where you cut it so you can get a good picture.  Good luck with that.

I guess workman knows how to find pleurocystidia in a dried sample and maybe he has some insight into the problem.

Maybe the sample could be rehydrated and then cut in a similar way.  That would turn the sample into a blob of goo but maybe it would work.

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: Alan Rockefeller]
    #11137842 -

Thanks for the info Mr. Rockefeller!


--------------------
:macrofunction:

Mushroom Hunting and Identification Rules
Puget Sound Mycological Society
MushroomExpert.com
Sample ID Request - Necessary for proper ID's
An Overview on how to find Psilocybe mushrooms by Shockvalue
P. cinctulus (subbalteatus) vs. P. foenesecii
How do I put pictures in my posts?

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: Twiztidsage]
    #11137916 -

Amen. Thanks.

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: Alan Rockefeller]
    #11146507 -

Alan Rockefeller said:
Quote:
Is Ebay a good place to get a microscope? I am guessing no. If not, where?


Yes.  There are many suitable microscopes on ebay for good prices.  There are also lots of crappy microscopes and also really nice microscopes.  Sometimes for low prices.

This one costs $170 and has achromatic objectives:

http://cgi.ebay.com/BINOCULAR-BIOLOGICAL-MICROSCOPE-w-MECHANICAL-STAGE_W0QQitemZ350179455000QQcmdZViewItemQQptZLH_DefaultDomain_0?hash=item5188527018&_trksid=p3286.c0.m14

There are also many decent microscopes on amscope.com.

Quote:
(mycology) A relatively large cell found on the gill face of a basidiomycete


I wrote that. 

Its now the second google hit when you search for pleurocystidia.  I am so excited.  Its going to make me famous for sure.

Everyone that wants pleurocystidia will have to come to me to get them.  When putting a name on a species its really important to have them yet they can be really hard to find. 

Its easy enough to find them on fresh material - you cut the cap in an extremely thin slice perpendicular to the gills so you have a slice that looks like a very thin saw blade.  The pleurocystidia is on the sides of the saw teeth and the cheilocystidia on the tips. 

But when its dried its a lot harder to find the cells on the gill face.  You can't cut the cap like that, it will just break. Cheilocystidia is no problem.  For the pleurocystidia I guess you can rehydrade and fold the gill over.  But that only gives a narrow section of the gill face and usually there are no pleurocystidia there.  Also the gill usually breaks when you try to fold it over which pretty much sucks.

Maybe the key is to make lots of really thin slices of a bit of gill, and hope that some pleurocystidia shows up exactly where you cut it so you can get a good picture.  Good luck with that.

I guess workman knows how to find pleurocystidia in a dried sample and maybe he has some insight into the problem.

Maybe the sample could be rehydrated and then cut in a similar way.  That would turn the sample into a blob of goo but maybe it would work.


Yeah, that's the link I was talking about.

And yeah, thanks for posting all of that.  I was just thinking we should share a few more tips on microscopy here for those interested in it.

When I am looking for pleurocystidia from a dried specimen I rehydrate a gill with 3% KOH solution and lay it flat on a slide.  Then I use a razor blade and slice off the thinnest slices I can.  When I have a few of them I put a cover glass over the mess and start looking around.  Most of the time I can find them. Photographing them is my problem.  I use a camera with a smaller lens than the ocular like you suggested.  Still I have problems focusing, even more problems holding the camera steady.  I think I might have a holder special made for it.

Maybe Workman will share his method with us like you said.


--------------------

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: Mr. Mushrooms]
    #11146709 -

I pretty much do the same thing as Mr. Mushrooms on the slicing, but I only put the best slice on the slide with a drop of KOH solution.  I then press it down with a pencil eraser to flatten the gill slice and expell any air bubbles.  You have to mash it enough to get good visibility but not so much that you destroy the sample.  It often takes a few trys.

I've found that the less material on the slide, the sharper the image.  There is something about extra material on the slide, even if its not directly under the objective, that muddies the image.


--------------------
Research funded by the patrons of
The Spore Works
Exotic Spore Supply

My Instagram
Reinvesting Sales Towards Basic Research and Species Identification :amanitajar:

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: Workman]
    #11146741 -

Sweet.  I'll have to try that.  I was afraid of getting the section on its side so I used a few of them.  If you mash it like you do that should solve that problem.


--------------------

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: Mr. Mushrooms]
    #11146878 -

If your slice is thinner than the gill is wide it naturally floats to the right orientation (in theory) :wink:


--------------------
Research funded by the patrons of
The Spore Works
Exotic Spore Supply

My Instagram
Reinvesting Sales Towards Basic Research and Species Identification :amanitajar:

Extras: Filter Print Post Top
Re: Calibrating Microscope [Re: Mr. Mushrooms]
    #11146888 -

I think the problem with too much material on the slide is that it is too wide creating a greater depth of focus, hence the importance of creating as thin a section as possible, it takes practice.
A good way of telling if your sections are thin enough is if your slide cover sits flat against your slide when applied!
A problem with the crush mount technique is the distortion of the cells which in turn gives you inaccurate measurements.
I have worried about the same thing you describe Mr. Mushrooms where the section tips on it's side, this is another reason for the importance of as thin a section as possible!

Workman, do you have any microscopy images of the basidia of penis envy, I would be very interested in seeing some!
inski..


--------------------

Edited by inski (09/28/09 09:35 PM)

Extras: Filter Print Post Top
Jump to top Pages: 1 | 2 | 3 |  [ show all ]

Shop: Mushroom-Hut Substrate Bags   Myyco.com Isolated Cubensis Liquid Culture For Sale   Sporeworks.EU Spores for European Microscopy   North Spore North Spore Mushroom Grow Kits & Cultivation Supplies

Mushrooms, Mycology and Psychedelics > Mushroom Hunting and Identification
Threaded Mode Threaded  Previous topic Previous  View all topics Index  Next topic Next 

Similar ThreadsPosterViewsRepliesLast post
* Notes on microscopy and how to get started
( 1 2 3 all )		 The Lightning 8,247 55 05/13/12 09:33 AM
by The Lightning
* Psilocybe stuntzii look-alike sp. microscopy (pleurocystidia with explanation)
( 1 2 all )		 The Lightning 5,835 32 11/16/11 06:23 PM
by psylosymonreturns
* microscope leafing 2,040 8 03/19/05 09:44 AM
by kadakuda
* What kind of microscope
( 1 2 all )		 tonyshroom 6,995 33 02/23/11 01:49 PM
by BlimeyGrimey
* how do you tell the species of a shroom using a microscope?
( 1 2 3 all )		 robocopdead 3,791 44 06/14/12 07:25 PM
by Alan Rockefeller
* Eastern North America Request - UPDATE with microscopy Eastern Baeocistis
( 1 2 all )		 rope 3,136 28 05/11/14 04:33 PM
by rhizoRider
* Pan foe microscopy pics. NeoSporen 650 7 06/07/11 10:28 PM
by sporeRider
* I need potential microscope owners to help with an FAQ
( 1 2 all )		 The Lightning 2,606 27 12/07/12 06:00 AM
by NeoSporen

Extra information
You cannot start new topics / You cannot reply to topics
HTML is disabled / BBCode is enabled
Moderator: ToxicMan, karode13, inski, Alan Rockefeller, Duggstar, TimmiT, Anglerfish, Tmethyl, Lucis, Doc9151, Land Trout
6,331 topic views. 0 members, 59 guests and 12 web crawlers are browsing this forum.
[ Show Images Only | Sort by Score | Print Topic ]
Search this thread:

Copyright 1997-2026 Mind Media. Some rights reserved.

Generated in 0.038 seconds spending 0.01 seconds on 18 queries.