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 gsharpnolack
Registered: 09/05/11
Posts: 657
Loc: Washington State US 
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 Re: update to microscopy notes [Re: suchen]
#15808552 - 02/14/12 01:57 PM (3 months, 12 days ago) |
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I'm not familiar with water immersion. I use an achromatic plan objective (60x) just like I use my 40x objective.
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One thing that can improve our planet: Personally creating a small amount of special atmosphere within the home place.
http://en.wikipedia.org/wiki/Four_Noble_Truths
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RigVedaLXVII.Soma
Bacteri/Phyc/Bry/Myc/ology
Registered: 11/21/11
Posts: 380
Loc: Michigan
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Re: update to microscopy notes [Re: suchen]
#15808616 - 02/14/12 02:10 PM (3 months, 12 days ago) |
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Aha, I see. But Wouldn't it be beneficial to give the section more depth of field, or are you saying it is too overkill for this purpose?
As shown in this image, even though it isnt a gill section, it isnt too much thicker than a gill section. C. in the figure is was captured with a hoffmann phase contrast and water immersion lens.
Either way, I think I could find many uses for it besides strictly fungal microscopy. 
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     "While clinging to the moronic belief that they constitute a "counterculture," they share our society's overriding urge for expediency. They make no attempt to learn about the organisms they eat and it always struck me as ironic that people with such a low level of consciousness should be seeking "higher consciousness." -David Arora
"Man is placed in the middle between two infinities - the infinitely great and the infinitely little - both of which are equally incomprehensible to him." -Pascal
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 Terry M
Stranger in a Strange Land
Registered: 06/18/10
Posts: 800
Loc: Rhode Island
Last seen: 4 hours, 12 minutes
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Re: update to microscopy notes [Re: suchen]
#15808633 - 02/14/12 02:14 PM (3 months, 12 days ago) |
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Great info in general. I like the stain step-by-step, because I'm just a beginning microscopist and haven't seen the mounting and staining process shown in this much detail before. Got my Congo Red on the shelf and ready to go.
I also found it particularly interesting about observing a gill rather than a gill cross section. This looks like a nice quick technique. I still want to practice cutting cross sections, though. I've only cut my thumb once so far!
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Obsessed with edibles all my waking hours.
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hungrygoldfish
Registered: 10/23/10
Posts: 641
Loc: UK 
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Re: update to microscopy notes [Re: Terry M]
#15819072 - 02/16/12 01:28 PM (3 months, 10 days ago) |
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I contacted the seller I got my microscope from to buy some other bits for it and about the 60x objective he said:
Quote:
The 60x objective will work on your scope but you need to re-focus as it is not parfocal with your Kyowa objectives. That 60x is a DIN type and is longer than yours.
What does that mean? Is it difficult to do?
Also in terms of measuring spores would it be better to get a Calibration slide or Eyepiece graticule or both?
Also for the oil lens is This the right type of oil?
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gsharpnolack
Registered: 09/05/11
Posts: 657
Loc: Washington State US
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Quote:
hungrygoldfish said:
I contacted the seller I got my microscope from to buy some other bits for it and about the 60x objective he said:
Quote:
Quote:
The 60x objective will work on your scope but you need to re-focus as it is not parfocal with your Kyowa objectives. That 60x is a DIN type and is longer than yours.
What does that mean? Is it difficult to do?
He's just saying that after you rotate from a less-powered objective (ie 10x) into the 60x you'll have to use the focus knobs to refocus. Piece of cake 95 percent of the time.
Quote:
Also in terms of measuring spores would it be better to get a Calibration slide or Eyepiece graticule or both?
For measuring spores you can get this: http://store.amscope.com/ep10x23r.html or another similar one
Quote:
Also for the oil lens is This the right type of oil?
This is what I use for immersion oil:
Your stage micrometer will work.
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One thing that can improve our planet: Personally creating a small amount of special atmosphere within the home place.
http://en.wikipedia.org/wiki/Four_Noble_Truths
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RigVedaLXVII.Soma
Bacteri/Phyc/Bry/Myc/ology
Registered: 11/21/11
Posts: 380
Loc: Michigan
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yeah man, Idk If I would buy immersion oil from the link you posted. They list it in Pet Health/care section, and have NO description whatsoever.
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"While clinging to the moronic belief that they constitute a "counterculture," they share our society's overriding urge for expediency. They make no attempt to learn about the organisms they eat and it always struck me as ironic that people with such a low level of consciousness should be seeking "higher consciousness." -David Arora
"Man is placed in the middle between two infinities - the infinitely great and the infinitely little - both of which are equally incomprehensible to him." -Pascal
MyPostImage Gallery Trade ListSpores I Am Looking For Seeds I am Looking For Solanaceae, Amanitaceae
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Byrain
-
Registered: 01/07/10
Posts: 1,819
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Cargille is great.
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hungrygoldfish
Registered: 10/23/10
Posts: 641
Loc: UK
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Re: update to microscopy notes [Re: Byrain]
#15819710 - 02/16/12 04:11 PM (3 months, 10 days ago) |
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Quote:
gsharpnolack said:
He's just saying that after you rotate from a less-powered objective (ie 10x) into the 60x you'll have to use the focus knobs to refocus. Piece of cake 95 percent of the time.
I have to always refocus anyway!
Quote:
RigVedaLXVII.Soma said:
yeah man, Idk If I would buy immersion oil from the link you posted. They list it in Pet Health/care section, and have NO description whatsoever.
That's exactly why I was asking, I have a £20 amazon voucher to use up so I was hoping that this product would be good 
Quote:
Byrain said:
Cargille is great.
Thanks for the tip!
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TimmiT
Registered: 03/23/10
Posts: 2,431
Loc: Victoria 
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Quote:
What does that mean? Is it difficult to do?
I've never had to add an objective, so I'm not sure what's involved. I imagine that it would be fairly straight forward though. He means what gsharpnolack said... that you will have to refocus when changing objectives. In an ideal world when you change objectives the subject will remain in focus because they're made so that the focal distances match. More often the focus will need fine adjustment. If you used that 60X the focus would not match the other objectives and it would take more than fine adjustment
Quote:
Also in terms of measuring spores would it be better to get a Calibration slide or Eyepiece graticule or both?
You need both. An eyepiece with a reticle (probably easier to buy an eyepiece with the reticle already installed) and a stage micrometer (calibration slide). You will only need the stage micrometer once so try borrowing one if you can. It's purpose is to measure the distance between the graduations of the reticle. Once you know the distances for each objective you don't need the micrometer again.
Quote:
Also for the oil lens is This the right type of oil?
That oil looks fine.
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"Reality leaves a lot to the imagination" ~ John Lennon
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Terry M
Stranger in a Strange Land
Registered: 06/18/10
Posts: 800
Loc: Rhode Island
Last seen: 4 hours, 12 minutes
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Re: update to microscopy notes [Re: TimmiT]
#15822767 - 02/17/12 09:16 AM (3 months, 10 days ago) |
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There are Type A and Type B of immersion oil commonly used. I asked a pathologist friend what kind he uses. He said a mixture of both types, though he didn't know the proportions used in his hospital because someone in his lab mixes them for him.
Type B is more viscous than Type A. I bought both types, but so far I've just used Type A, which appears to work fine as far as I can tell.
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Obsessed with edibles all my waking hours.
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gsharpnolack
Registered: 09/05/11
Posts: 657
Loc: Washington State US
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Re: update to microscopy notes [Re: Terry M]
#15833648 - 02/19/12 05:34 PM (3 months, 7 days ago) |
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Updated above
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One thing that can improve our planet: Personally creating a small amount of special atmosphere within the home place.
http://en.wikipedia.org/wiki/Four_Noble_Truths
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RigVedaLXVII.Soma
Bacteri/Phyc/Bry/Myc/ology
Registered: 11/21/11
Posts: 380
Loc: Michigan
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I was given the permission of my very kind friend gsharpnolack to post my relevent thread, "DIY Microtome; For those thin sections you just cant get yourself" in hopes that some may find it useful in sectioning, and so that the new people who gain interest in microscopy dont lose hope or aspiration just because they cant get thin sections by hand. Now everyone can get almost paper thin sections with ordinary houshold items. No reason that one cant get at least 10 micron thin sections.
Thank you gsharpnolack. I appreciate your kindness.
And dont give up folks! Whether you try my DIY microtome tek, or section by hand, DO NOT GIVE UP! Practice Practice Practice!!
Happy Microscopy!!
Good Luck!
There is a whole other world under that scope!
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"While clinging to the moronic belief that they constitute a "counterculture," they share our society's overriding urge for expediency. They make no attempt to learn about the organisms they eat and it always struck me as ironic that people with such a low level of consciousness should be seeking "higher consciousness." -David Arora
"Man is placed in the middle between two infinities - the infinitely great and the infinitely little - both of which are equally incomprehensible to him." -Pascal
MyPostImage Gallery Trade ListSpores I Am Looking For Seeds I am Looking For Solanaceae, Amanitaceae
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Alan Rockefeller
Mycologist
Registered: 03/10/07
Posts: 24,722
Last seen: 4 days, 19 hours
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Re: update to microscopy notes [Re: Terry M]
#15835939 - 02/20/12 12:42 AM (3 months, 7 days ago) |
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Quote:
Terry M said:
There are Type A and Type B of immersion oil commonly used. I asked a pathologist friend what kind he uses. He said a mixture of both types, though he didn't know the proportions used in his hospital because someone in his lab mixes them for him.
Type B is more viscous than Type A. I bought both types, but so far I've just used Type A, which appears to work fine as far as I can tell.
I like type A because when I use type B, it has a tendancy to slide the cover slip over when I move the objective away.
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gsharpnolack
Registered: 09/05/11
Posts: 657
Loc: Washington State US
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I'm trying to find an ideal stain for specifically working with dried/rehydrated gilled mushrooms and found these stains:
http://www.mycology.adelaide.edu.au/Laboratory_Methods/Microscopy_Techniques_and_Stains/
Do any of you have an opportunity to try one of these out and share your results?
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One thing that can improve our planet: Personally creating a small amount of special atmosphere within the home place.
http://en.wikipedia.org/wiki/Four_Noble_Truths
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Alan Rockefeller
Mycologist
Registered: 03/10/07
Posts: 24,722
Last seen: 4 days, 19 hours
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My favorite stain is cotton blue. It is extremely blue.
I wonder how well food coloring would work.
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TimmiT
Registered: 03/23/10
Posts: 2,431
Loc: Victoria
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The Adelaide Uni site covers medical mycology, so the techniques and stains are more relevant to that field. You probably won't find them used outside a diagnostics lab.
I have pics of PAS and silver stains (and a few other histo stains) in my gallery if you want to see what they look like (not fungal though).
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"Reality leaves a lot to the imagination" ~ John Lennon
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bloodworm
cube con·nois·seur
Registered: 05/22/10
Posts: 4,073
Loc: 352
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Re: update to microscopy notes [Re: TimmiT]
#15846650 - 02/22/12 08:23 AM (3 months, 5 days ago) |
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i found an excellent stain.
ink stamp refills work awesome...and they are cheap (you can find them at any hobby or office store).
here are some pics where i used ink stamp refills as the stain.
  
Pluteus cervinus cystidia @ 400x in a crush mount
just be careful not to get any of the ink on your lenses.
peace and love
bloodworm
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Alan Rockefeller
Mycologist
Registered: 03/10/07
Posts: 24,722
Last seen: 4 days, 19 hours
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Re: update to microscopy notes [Re: bloodworm]
#15849378 - 02/22/12 06:52 PM (3 months, 4 days ago) |
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Quote:
Pluteus cervinus cystidia @ 400x in a crush mount
Thats not a crush mount. Or if it is, it isn't crushed enough to free the cells.
In a crush mount you should be able to see the whole cell, not just the part that is sticking out of the gill.
The stain looks really good, but I wonder why there is an aura around the cystidia. I have never seen that before.
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falcon
In the green
Registered: 04/01/02
Posts: 5,150
Last seen: 20 hours, 18 minutes
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Thanks!
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gsharpnolack
Registered: 09/05/11
Posts: 657
Loc: Washington State US
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Quote:
In a crush mount you should be able to see the whole cell, not just the part that is sticking out of the gill.
Can you share a few examples?
--------------------
One thing that can improve our planet: Personally creating a small amount of special atmosphere within the home place.
http://en.wikipedia.org/wiki/Four_Noble_Truths
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