First attempt at a thin section with a manual microtome

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Macro_Cosmos
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First attempt at a thin section with a manual microtome

Post by Macro_Cosmos »

Cheap manual slicing microtome came in. Snipped some stems from the garden.
Jammed the stem into a carrot and sliced with a razor blade.
carrot.jpg
Slices aren't thin enough. Claims to do 20um, best I can get is probably 50um. The microtome does feel flimsy, perhaps I can get better with practice and using wax/paraffin instead of innocent carrots minding their own businesses before I cook them. Brunel offers a nice looking one which boasts 2um resolution? Has anyone actually used it?
https://www.brunelmicroscopessecure.co. ... tomes.html
Bench microtome found in the above link is the one I was referring to.

Not bad, but could do well with a thinner slice.
Image

Luis Barrera Puigdollers
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Re: First attempt at a thin section with a manual microtome

Post by Luis Barrera Puigdollers »

Hi, In my humble opinion, a very thin section is useful for botanists but lacks the beautiful 3D look for a considered photogenic image. I've tried with embedded paraffin and a lab microtome ,and sections over 20 microns usually roll up on themselves making them useless, but is not impossible. Not all botanical material has the same hardness even within the same piece and you need to test. With a manual microtome sections of already 50 microns or so, look well but practice is necessary to a uniform cut. With very hard material its another history and you have a lot of bibliography to complicate a bit your life. I strongly recommend the manual microtome. Look the slides of C. Krebs for example

Luis Barrera Puigdollers
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Re: First attempt at a thin section with a manual microtome

Post by Luis Barrera Puigdollers »

Hi again,
With a manual microtome, the blade and "cutting art" are critical. I never use the blade standard of a microtome. I use a razor blade mounted on a thick handle like glass scrappers to best control

Macro_Cosmos
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Re: First attempt at a thin section with a manual microtome

Post by Macro_Cosmos »

Luis Barrera Puigdollers wrote:
Tue Feb 23, 2021 11:15 am
Hi again,
With a manual microtome, the blade and "cutting art" are critical. I never use the blade standard of a microtome. I use a razor blade mounted on a thick handle like glass scrappers to best control
Thanks for your info! Very helpful. I watched many videos on microtomes, so I try to level the blade at a good angle and made several successful cuts that were nice and thin-ish.

Does Krebs use the same Brunel model? His work is great. The Brunel model looks very well made, I need their Naphrax diatom mountant and maybe some other items, so might as well pick it up. Shipping is very expensive, so I just want to get all the stuff I need and end there. I do want to try staining some of these too, so around 25um would be great.

Thanks for the input!

Pau
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Re: First attempt at a thin section with a manual microtome

Post by Pau »

I've used this big scalpel blades with hand microtomes with good success when teaching Microscopy techniques at High School level:
https://www.swann-morton.com/product/33.php
Pau

Luis Barrera Puigdollers
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Re: First attempt at a thin section with a manual microtome

Post by Luis Barrera Puigdollers »

Hi,
Before buying a rotary microtome you should consider several things: the purpose of course (animal or plant tissue), what type of blade will you use ( the large blade with triangular shape difficult to sharpen and expensive or the thin disposable and cheap) what type of paraffin block (cube-shaped or with cassette included).I am not a pathologist but I have some experience making histological preparations. I strongly recommend the use of plastic cassettes to avoid manipulate tissues during the embedding process, and the use of thin disposable blades.Making paraffin blocks with tissues at home can be frustrating, you need a well ventilated space especially if you use xylene, where children do not enter (nor the pets), and an understanding wife or husband.I am lucky (or not) to live alone, to have enough space, and to have three microtomes that I pulled out of a garbage container almost 20 years ago. I usually use a Microm HM330.Fortunately I also have a gas mask and a house on the beach.
It is for these reasons that I recommend starting with a handheld microtome. For botanical preparations I use AAF as a fixative, distilled water, ethanol and isopropanol. And for staining fuchsin-safranin-astrablau of Morphisto. Mounting with Euparal (Carl Roth)

Luis Barrera Puigdollers
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Re: First attempt at a thin section with a manual microtome

Post by Luis Barrera Puigdollers »

Look the post "A Rose In Our Garden" (single shots)

Macro_Cosmos
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Re: First attempt at a thin section with a manual microtome

Post by Macro_Cosmos »

Luis Barrera Puigdollers wrote:
Wed Feb 24, 2021 9:50 am
Look the post "A Rose In Our Garden" (single shots)
Thanks again for the insight! I'm not going to deal with weird chemicals with odd symbols as of now, I need a decent space first and some kind of safe to store those nasty looking things. I'm not going to bother with heavy lab microtomes, they are forbiddingly expensive to ship, even if seemingly capable "AO Spencer Model 820" microtomes can be had at less than $200.

I'm wondering if anyone has any experience with the bench microtome shown here, for 40 pounds. https://www.brunelmicroscopessecure.co. ... tomes.html
I plan on using carrots and candle wax. I'll try and find a local source for the blade Pau pointed to.

That stained rose stem is beautiful. Coincidentally, I was reading details of the exact staining method the author used.
Robin Wacker 3A Staining method: https://moticeurope.blogspot.com/2021/0 ... er-3a.html

NikonUser
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Re: First attempt at a thin section with a manual microtome

Post by NikonUser »

The double-blade method works well. Will not always give an intact section, will do so for thick sections, but it will give a portion of the section that can be in the region of a few cells deep.
Equipment needed is cheap.
http://www.microscopy-uk.org.uk/mag/ind ... icerb.html
NU.
student of entomology
Quote – Holmes on ‘Entomology’
” I suppose you are an entomologist ? “
” Not quite so ambitious as that, sir. I should like to put my eyes on the individual entitled to that name.
No man can be truly called an entomologist,
sir; the subject is too vast for any single human intelligence to grasp.”
Oliver Wendell Holmes, Sr
The Poet at the Breakfast Table.

Nikon camera, lenses and objectives
Olympus microscope and objectives

Luis Barrera Puigdollers
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Re: First attempt at a thin section with a manual microtome

Post by Luis Barrera Puigdollers »

The bench microtome you mention should work very well. I never use a handheld microtome holding it with my left hand, so the cutting precision is not obtained in my opinion and it needs a clamping to a workbench, table or kitchen counter.

ChrisR
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Re: First attempt at a thin section with a manual microtome

Post by ChrisR »

Chris R

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