Home made (Very Basic) Microtome

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Roldorf
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Home made (Very Basic) Microtome

#1 Post by Roldorf » Mon Aug 19, 2019 2:40 pm

Hi Scouring the back posts from early 2016 I came across two posts by members one from JimT and one from rabbitt on their home made microtomes.

Combining the two ideas I am trying out my own version only not as sophisticated as using an old micrometer (as I don't have one :D ).

Hang fire first I will show you what I did. (It still hasn't cooled off, hardened, yet) and I still have to trim the whole thing and find a way to mount it on the bench. Clamping the sample still has to be worked out probably a carrot or some pith from an elderberry branch.

Tools and materials:-

Wood saw (to saw the wood).
Drill and drill bits (to drill the wood).
Pen
Wood (piece of scrap lying around)
Round wooden broom handle (Old) ;)
Araldite.
Bolt (suitable size to fit the hole)
Washers (to fit the bolt)
Nut (to fit the bolt)
Aluminium tube (found in a bits box don't know what it's off) but it fits bolt.
A big washer with a small hole for the cutting surface (also from the bits box)

OH and last but not least some Knife blades (New but had them anyway)
Lots of patience. :shock:

Ok here it is:-

Image

Image

Image

I am hoping that the rudimentary markings on the 'Knob' will give me somewhere in the region of 0.018mm sections ;)
Alan
Bresser Science Infinity 4x 10x 40x 60x 100x oil. Canon EOS 4000d
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Roldorf
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Re: Home made (Very Basic) Microtome

#2 Post by Roldorf » Mon Aug 19, 2019 6:17 pm

First Slides/Images:-

4x Horsemint Stem Crossection


Image

10X


Image


I can see i need more practice. Raw images no alterations as they were cut.

I had to pack the hole with lots of the stems so that i could take a slice. Done with a carpet knife. No cover-glass.

Considering I am very happy with my very first try. :D


Errr by the way I have no idea what I am looking at. :roll:
Last edited by Roldorf on Mon Aug 19, 2019 9:25 pm, edited 1 time in total.
Alan
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MicroBob
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Re: Home made (Very Basic) Microtome

#3 Post by MicroBob » Mon Aug 19, 2019 6:26 pm

Hi Alan,
this will probably work acceptably well! When using a cylinder microtome like this it is best to use a blade holder that leads the blade at the right inclination over the microtome without the edge touching anything else than the object. The SHK holder is an example for this. I designed a 3D-printed blade holder for the especially sharp OLFA LBB blades: https://www.amazon.de/OLFA-10St%C3%BCck ... B0029F8RDQ
To bed the object it is best to embed it in two pieces of carrot. One section is the result of two cuts. The more uniform you do these, the more even the section. Here you can find a couple of hints: viewtopic.php?f=16&t=7065

Bob

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Re: Home made (Very Basic) Microtome

#4 Post by Dave S » Mon Aug 19, 2019 6:36 pm

Very innovative Alan, well done.

As the workman said to the lady who asked "how do I get to Carnegie Hall", "you have to practice lady" he said :)

From what I have seen, and as Bob suggests, using a Carrot to hold the sample does seem to be a popular method. Certainly got to be a lot easier than wax embedding.
Brunel SP100 (with 4x, 10x, 40x,60x, and 100x (oil) plan objectives), and Canon EOS 4000d Camera (microscopy use only)

Roldorf
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Re: Home made (Very Basic) Microtome

#5 Post by Roldorf » Mon Aug 19, 2019 6:40 pm

Thanks guys I will have to steal some carrot's from my wife's kitchen supplies. :twisted:
Alan
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Re: Home made (Very Basic) Microtome

#6 Post by MicroBob » Mon Aug 19, 2019 6:58 pm

On the MKB Bonn website you can probably find the most valuable collection of information on plant sectioning for the amateur: http://www.mikroskopie-bonn.de/biblioth ... index.html
They also show how to dress the carrot herev.
With these methods it is comparatively easy to obtain nice plant section slides with differentiated staining. I'm doing this only occasionally but got nice results here: viewtopic.php?f=6&t=7013

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mrsonchus
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Re: Home made (Very Basic) Microtome

#7 Post by mrsonchus » Mon Aug 19, 2019 8:45 pm

This looks like a very nice start to me, nicely-done!
Good clear image showing lots of anatomy.
John B

billbillt
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Re: Home made (Very Basic) Microtome

#8 Post by billbillt » Mon Aug 19, 2019 11:05 pm

Hello,

This is a good idea.. Where did you come up with your divisions?.. Did you take the pitch of the screw into account?.. The bolt you are using looks fairly coarse.. a finer pitch nut/bolt combination would allow for thinner sections...

The Best,
BillT

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KurtM
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Re: Home made (Very Basic) Microtome

#9 Post by KurtM » Tue Aug 20, 2019 1:09 am

Very cool!

Just in case somebody finds it useful, here's a hint for the sharpest razor blades you can buy locally: They still make and sell double-edged safety razor blades (I'm in the USA), find 'em in the grocery store, much keener than box cutter or single edge razor blades. Super glue or tape a blade to a popsicle stick or similar, and you have a heck of a cutting instrument. But do beware it's a "double edge sword". I actually fold them over until they snap in half, and glue a half-blade to a stick. Of course you want to be careful doing it, wear eye protection, but you ought to be observing meticulous safety protocol in the lab anyway, right?

Back when I was building and flying F1D Indoor Free Flight, I learned to dissect Gillette Mach 3 razor heads for the individual blades, which I super-glued to stick handles, to cut microfilm for wings and tail surfaces. Tiny little things are a bit of trick to handle, but even sharper yet.

PS: In case you're wondering what F1D Indoor Free Flight is: https://www.youtube.com/watch?v=sVaDvS33Z5Q
Cheers,
Kurt Maurer
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bobm4360
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Re: Home made (Very Basic) Microtome

#10 Post by bobm4360 » Tue Aug 20, 2019 4:03 am

Single-edged razor blades can be purchased for shaving at the pharmacy, and they are considerably sharper than the ones you buy for craft, box cutters, etc.

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Re: Home made (Very Basic) Microtome

#11 Post by Hobbyst46 » Tue Aug 20, 2019 7:56 am

KurtM wrote:...PS: In case you're wondering what F1D Indoor Free Flight is: https://www.youtube.com/watch?v=sVaDvS33Z5Q
Thanks for the link! hey, the indoor slow-motion hoverfly is at least as attractive as microscopy!!
Zeiss Standard GFL+Canon EOS-M10, Olympus VMZ stereo

Roldorf
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Re: Home made (Very Basic) Microtome

#12 Post by Roldorf » Tue Aug 20, 2019 10:30 am

Hi billbillt

I measured the pitch (1.3 mm by turning the nut one revolution) with a vernier caliper. Using the bolt head which I had attached to the wooden broom handle, I divided the circumference into 12 (flats and corners) then divided the marks in two to get 24 divisions, then it was pitch divided by 24 to get the movement for 1 division. A bit rough and ready, but the whole thing is anyway and it can't be too far out.

It was the finest thread that I had that would fit through the aluminium sleeve so that made my decision easy.
Last edited by Roldorf on Wed Aug 21, 2019 3:37 pm, edited 1 time in total.
Alan
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Re: Home made (Very Basic) Microtome

#13 Post by Roldorf » Tue Aug 20, 2019 10:39 am

Hi KurtM, bobm4360,

Thanks for the information on blades. I have regular old fashioned razor blades that we use for sectioning mushrooms. We usually use two together by snapping a blade in half holding the two half's pressed together then making the cut through the cap/stem or whichever part of the mushroom we want to look at under the scope. The very thin slice can be removed from between the two blades. Works well but only on very soft materials, hence my try at making a microtome.

I will try the carrot trick later today.
Alan
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Re: Home made (Very Basic) Microtome

#14 Post by tgss » Tue Aug 20, 2019 2:16 pm

KurtM wrote: Back when I was building and flying F1D Indoor Free Flight, I learned to dissect Gillette Mach 3 razor heads for the individual blades, which I super-glued to stick handles, to cut microfilm for wings and tail surfaces. Tiny little things are a bit of trick to handle, but even sharper yet.
Nice to know there are other ex-aeromodellers on the forum!

Tom W.
Class A combat, class A team race, in the UK before the current FAI classes were formulated! Latterly went to the dark side AKA radio control.
Now back on topic.

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Re: Home made (Very Basic) Microtome

#15 Post by Roldorf » Fri Aug 23, 2019 12:34 pm

Latest test from my microtome also a test once more of google photos according to mrsonchus.
Here we go:-
Image
Last edited by Roldorf on Fri Aug 23, 2019 12:42 pm, edited 1 time in total.
Alan
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Re: Home made (Very Basic) Microtome

#16 Post by MicroBob » Fri Aug 23, 2019 12:40 pm

Hi Alan,
this is starting to look really good!
For further improvent you might look into stacking. Picolay is free and works well.
To my surprise stacking still improves the quality of images of flat sections - the depth of focus is smaller than the slight unevenness of the sections. This will get rid of the slightly hazy areas in the image.

Bob

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Re: Home made (Very Basic) Microtome

#17 Post by Roldorf » Fri Aug 23, 2019 12:47 pm

Hi Bob, sorry to disappoint but this was stacked Oh no. I stacked it with Photoshop, I usually try both PS and Picolay and choose the result that looks the best, in this case it was PS. I think I could probably get it better with Picolay but there are so many stacking options I can never seem to get the best one.
Alan
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Re: Home made (Very Basic) Microtome

#18 Post by Roldorf » Fri Aug 23, 2019 12:56 pm

I have found that when sectioning it seems as though turning the screw for the next cut seems to result in a delay, possibly caused by compression of the sample.
Turn screw to raise sample, no section possible. turn screw again very thick section.
Wait a short while try section again, very thin section.
So it seems to be turn screw, wait a minute or two take section.
Alan
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Re: Home made (Very Basic) Microtome

#19 Post by MicroBob » Fri Aug 23, 2019 2:04 pm

I found it to be important to choose the right images to stack and leave all other images out. So may be begin 1 image before the first sharp areas appear, then through the stack down to the middle of the object or so, not too deep. In Picolay you can select "align images". I'm not a stacking expert and probably can't really help here.

The problem with the varying thickness is common. Here a couple of causes I have experienced:
- Thickness of set cut too set shallow for the type of material. The Sansevieria needed between 70 and 100µ to be cut properly
- Edge not sharp or folded over from contact to the microtome, raises the minimum possible thickness
- Motion not uniform from cut to cut
- It is best to move the blade along the stem until you see it enters it - then continue the cut right trough (I have tried to show the motion in my manual for the blade holder)

Bob

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Re: Home made (Very Basic) Microtome

#20 Post by Roldorf » Sat Aug 24, 2019 10:18 am

I have just completed 'Home Made Microtome version 2'.
It has a larger screw but finer thread and has a much more depth for specimens also a wider area for the blade. I hope that this will maybe alleviate some of the twisting that I have observed when making cuts with the blade. A test later this afternoon with the same material as before (when the Araldite has gone off properly).
Alan
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Re: Home made (Very Basic) Microtome

#21 Post by Roldorf » Sat Aug 24, 2019 11:28 am

Ok same plant stem as before with Microtome Ver 2.
I found it much easier to get a fine section with the new version.
Image is full size not edited apart from an 8 image stack and a bit of enhanced brightness and contrast. This stack is also with Photoshop.

Image
Alan
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Re: Home made (Very Basic) Microtome

#22 Post by Roldorf » Sat Aug 24, 2019 4:15 pm

Ok Version 2 will be the one I use from now on Ver 1 is hereby retired.

Managed to get a very fine slice with the new and improved Ver 2. Same plant stem as before.

Image

For those of you who are interested here is a drawing and some pics of the beast:-
rect5960.png
rect5960.png (17.07 KiB) Viewed 2463 times

Image

Image

Image

This last image is of the washers used to line the sample bore 29,8 x 12mm.
Alan
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Re: Home made (Very Basic) Microtome

#23 Post by mrsonchus » Sat Aug 24, 2019 5:34 pm

Looks good Roldorf, sections are steadily improving!

Interested to see how you progress with them using the v2, and with differing stem-types.

Nice work my friend, keep 'em coming. :)
John B

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Re: Home made (Very Basic) Microtome

#24 Post by tgss » Sat Aug 24, 2019 6:32 pm

Good job all round Alan! Pretty soon John B is going to have to look to his laurels ;)
Tom W.

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Re: Home made (Very Basic) Microtome

#25 Post by MicroBob » Sat Aug 24, 2019 7:19 pm

Hi Alan,
to me these sections look even but a bit thick. One point of improvement might be the surface of the top washer. As it looks it might easily scratch the edge while cutting. I haven't tested them for this purpose but the discs in hard discs are even, flat, have a smooth surface, a hole in the middle and are cheap.

Bob

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Re: Home made (Very Basic) Microtome

#26 Post by Roldorf » Mon Aug 26, 2019 2:44 pm

Tried using carrots. I have found that trying to get the stem to cut correctly in the middle of the carrot is a pain.
I find the easiest way is to put the whole carrot in (at just less than the size required) and then pushing the sample you wish to cut to the back makes it very easy to do.


Image
Last edited by Roldorf on Thu Aug 29, 2019 2:04 pm, edited 1 time in total.
Alan
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Re: Home made (Very Basic) Microtome

#27 Post by Roldorf » Thu Aug 29, 2019 2:02 pm

Can't see from this 10x section but the runner is square. Quick Section:-

Image
Alan
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Re: Home made (Very Basic) Microtome

#28 Post by MichaelG. » Mon Oct 07, 2019 3:42 pm

Roldorf,

Now that you have proved you can make your own ...
You may be interested in this; which is currently on special offer
https://www.surplusshed.com/pages/item/M1571D.html

Seems a fair price, and Fred is an honourable chap when it comes to postal charges.

MichaelG.
Too many 'projects'

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Re: Home made (Very Basic) Microtome

#29 Post by MicroBob » Mon Oct 07, 2019 3:58 pm

Hi Michael,
in my experience hand microtoms with object clamp and a big opening in the table are more useful. The shown has only a tiny hole in the table so you might be able to lead a stem through it, but not to support it on it's way.
Here in Germany hand microtomes are very popular, but they only work well with a blade holder that leads the blade above the table. These are common here in Germany but are not widely sold outside of the german speaking countries. This has probably contributed to a very weak supply of good hand microtomes.
With a blade holder they work like a charm for plant sections: viewtopic.php?f=16&t=7065&p=62601&hilit=blade#p62601

So I have started to think about a design for a modernized hand microtome that works better than a really good cylinder hand microtome and can be made readily available. Price range will probably be 150-200€. Progress depends on a lot of other projects so I can't promise then I can show more.

Bob

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Re: Home made (Very Basic) Microtome

#30 Post by MichaelG. » Mon Oct 07, 2019 6:35 pm

MicroBob wrote:
Mon Oct 07, 2019 3:58 pm
Hi Michael,
in my experience hand microtoms with object clamp and a big opening in the table are more useful. The shown has only a tiny hole in the table so you might be able to lead a stem through it, but not to support it on it's way.
Here in Germany hand microtomes are very popular, but they only work well with a blade holder that leads the blade above the table. […]
That’s interesting, Bob ...

The SurplusShed item looks to have very similar dimensions to my vintage [all Brass] one; which is, in turn, very similar to the ones we used at school more than 50 years ago. Both of these, of course, being used with a specially-ground cut-throat Razor.

I would be interested to see a photo of these currently popular modern German versions.

MichaelG.
Too many 'projects'

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