Hand sectioning; plants

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Rudix
Posts: 90
Joined: Sat Jan 21, 2017 11:30 am
Location: South Africa

Re: Hand sectioning; plants

Post by Rudix »

NikonUser wrote:
Fri Mar 26, 2021 12:42 pm
A further, final, comment on hand sectioning plant material; (originally posted on MicrobeHunter).

I have been trying several techniques to get cross-sections of plant stems including 'hand' cutting with the double razor blade, using a hand microtome fitted to a bench with the plant held by a foam plug and with the plant embedded in paraffin wax and using a cutting aid.
All techniques gave inconsistent results but the simplest method, by far, was the double razor blade. The simplicity allowed one to make dozens of sections in a very short time; usually one or two of the sections were suitable for further processing.
[my inconsistent, generally poor, results using the bench microtome and cutting aid was most likely due to not having the plant stem anchored firmly in the barrel of the microtome; the quality of the cutting aid and blade hold (from Germany) was excellent]

We bought some fresh carrots, all the way from California, which still had the greenery attached. I cut up a green stem and placed in 99% alcohol for 3 days. Cut with the double blade. Got quite a few decent complete sections. One partial section, last image, was about 1 cell thick!
Wow, fantastic results! Did you stain the sections?

Thanks for sharing, now I need to go find razor blades!
Rudi

NikonUser
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Joined: Thu Sep 04, 2008 2:03 am
Location: southern New Brunswick, Canada

Re: Hand sectioning; plants

Post by NikonUser »

Thanks.
Stained with Fast Green and Safranin-O
Perhaps a better method for sectioning:
https://pages.stolaf.edu/wp-content/upl ... 170415.pdf

OR:
The OPN Microtome: An Inexpensive,
Open Source Hand-Held Mini Microtome
John Giannini,* Associate Professor of Biology
St. Olaf College, 1520 St. Olaf Avenue, Northfield, MN 55057

Try again !
https://www.microbehunter.com/microscop ... opn#p74085
Last edited by NikonUser on Fri May 28, 2021 11:40 am, edited 1 time in total.
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

Rudix
Posts: 90
Joined: Sat Jan 21, 2017 11:30 am
Location: South Africa

Re: Hand sectioning; plants

Post by Rudix »

NikonUser wrote:
Fri May 28, 2021 11:31 am
Thanks.
Stained with Fast Green and Safranin-O
Perhaps a better method for sectioning:
https://pages.stolaf.edu/wp-content/upl ... 170415.pdf

OR:
The OPN Microtome: An Inexpensive,
Open Source Hand-Held Mini Microtome
John Giannini,* Associate Professor of Biology
St. Olaf College, 1520 St. Olaf Avenue, Northfield, MN 55057
Thanks for the prompt reply! Will go check the PDF in the link!
Rudi

rjlittlefield
Site Admin
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Re: Hand sectioning; plants

Post by rjlittlefield »

NU, thanks for the reference -- very interesting! (I admin-edited the post to fix a glitch in the URL, but then I see you did the same thing.)

--Rik

NikonUser
Posts: 2693
Joined: Thu Sep 04, 2008 2:03 am
Location: southern New Brunswick, Canada

Re: Hand sectioning; plants

Post by NikonUser »

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

Rudix
Posts: 90
Joined: Sat Jan 21, 2017 11:30 am
Location: South Africa

Re: Hand sectioning; plants

Post by Rudix »

The link worked fine thanks!

Interesting! I have downloaded the STL files and will print a copy tomorrow, looks like a nice little project that should be a nice addition to my equipment.

Appreciate the info, thanks!
Rudi

iconoclastica
Posts: 486
Joined: Sat Jun 25, 2016 12:34 pm
Location: Wageningen, Gelderland

Re: Hand sectioning; plants

Post by iconoclastica »

I just made a few of these. I had to change the design a little, for due to covid our printing workshop is still not running like it used to and I have to resort to the laser cutter. I made one with various sized holes to cut transverse sections and today I made another one with a horizontal slit for flat objects or longitudinal sections (see https://www.microbehunter.com/microscop ... 10&t=12939.

I was astonished to see how well this design works. I am a complete moron where it comes to cutting sections. I calculated today that I have failed to cut a decent section for 43 years - no joke.
But both of the microtomes I made, today and two weeks ago, produced usable thin cuts at the first try.
--- felix filicis ---

Rudix
Posts: 90
Joined: Sat Jan 21, 2017 11:30 am
Location: South Africa

Re: Hand sectioning; plants

Post by Rudix »

iconoclastica wrote:
Fri May 28, 2021 1:53 pm
I just made a few of these. I had to change the design a little, for due to covid our printing workshop is still not running like it used to and I have to resort to the laser cutter. I made one with various sized holes to cut transverse sections and today I made another one with a horizontal slit for flat objects or longitudinal sections (see https://www.microbehunter.com/microscop ... 10&t=12939.

I was astonished to see how well this design works. I am a complete moron where it comes to cutting sections. I calculated today that I have failed to cut a decent section for 43 years - no joke.
But both of the microtomes I made, today and two weeks ago, produced usable thin cuts at the first try.
Great news, thanks!

I am also looking at a few changes, I think it would be beneficial to make the gap between the slides adjustable, with a slight resizing of the print it should be possible to get a nice tight but not too tight fit between the printed part and the 2 slides allowing adjustment for thinner or thicker subjects. Might try to print the front section where the slides go with a different, more flexible material.

The basic design is simple so if I find the need I will just redo it from scratch in Fusion 360 or Solid Worx so that I have a file that it easier to edit. It is amazing how often I find things to print for my other hobbies, the printers also started as a hobby but they have become a fantastic tool.

Hope I can find razor blades today, I know I bought some a few years ago for this purpose but a major house move since then displaced a lot of stuff!

The method described to make sections of fragile subjects worry me a bit, once you have done the cut how do you remove the tape from the cut without damaging the section? Double sided tape normally has VERY strong adhesive, might have to resort to something to dissolve or at least soften the glue.
Rudi

iconoclastica
Posts: 486
Joined: Sat Jun 25, 2016 12:34 pm
Location: Wageningen, Gelderland

Re: Hand sectioning; plants

Post by iconoclastica »

Rudix wrote:
Fri May 28, 2021 9:15 pm
I am also looking at a few changes, I think it would be beneficial to make the gap between the slides adjustable, with a slight resizing of the print it should be possible to get a nice tight but not too tight fit between the printed part and the 2 slides allowing adjustment for thinner or thicker subjects. Might try to print the front section where the slides go with a different, more flexible material.
My front plate isn't fixed to the body and can be set into 3 positions, corresponding to 1mm, 2mm and 3mm holes that support the specimen in every direction (a second front plate additionally has 4mm and 5mm holes). You could try to place the two parts of the frontplate in the original design in a groove so you can slide them closer and wider.
Rudix wrote:
Fri May 28, 2021 9:15 pm
The basic design is simple so if I find the need I will just redo it from scratch in Fusion 360 or Solid Worx so that I have a file that it easier to edit.
In fact, when I made my version 1.0, I had placed the design files somewhere from where I couldn't download them once in the workshop. Then I made it completely anew in a couple of minutes just before cutting the acrylic.
Rudix wrote:
Fri May 28, 2021 9:15 pm
The method described to make sections of fragile subjects worry me a bit, once you have done the cut how do you remove the tape from the cut without damaging the section? Double sided tape normally has VERY strong adhesive, might have to resort to something to dissolve or at least soften the glue.
I place the whole device in a tray filled with so much water that the specimen is just in the water. Then I cut down, chac-chac-chac, as fast as I can, and only then start moving the specimen forward. The sections pile up at the bottom of the tray from where I pick them up with a needle.
--- felix filicis ---

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