Hand sectioning; plants

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

Hand sectioning; plants

Post by NikonUser »

In this month's Micscape Magazine there is an interesting technique:
http://www.microscopy-uk.org.uk/mag/indexmag.html

I have tried to hand cut sections using several methods, none worked! This method works at almost no cost.
Here is a section of a Hydrangea flower stem. I have only zoological stains which obviously don't work for botanical material :cry: . I have ordered botanical stains. I thought I would draw member's attention to the article while there is still time, for us in the north, to get plant material before winter.
Hopefully better images and interpretation after the stains arrive.
Nikon D7200, 4x CFN, Zerene PMax, flash; stem is 3.24mm wide
hydrangea 29x20.jpg
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


viktor j nilsson
Posts: 423
Joined: Fri Mar 01, 2013 1:43 am
Location: Lund, Sweden

Re: Hand sectioning; plants

Post by viktor j nilsson »

And for further reference, here's a link to the late Walter Dioni's original article:

http://www.microscopy-uk.org.uk/mag/artapr04/wdslicera.html

MarkSturtevant
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Joined: Sat Nov 21, 2015 6:52 pm
Location: Michigan, U.S.A.
Contact:

Re: Hand sectioning; plants

Post by MarkSturtevant »

This 'takes me back'. When I was in gradual school, my research involved doing lots of micro-surgery on various live insects and crustaceans. Legendary among us sorts were 'blue blades', which at the time were regarded as excellent quality. It has been a very very long time, but I suspect these are them: https://www.amazon.com/Silver-Double-Bl ... B0059QF5RS There is discussion in the comments about other brands that are also seen as exceptionally sharp.
Mark Sturtevant
Dept. of Still Waters

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

Re: Hand sectioning; plants

Post by NikonUser »

The only blades my pharmacy had were Wilkinson Sword, made in Germany. I bought a pack, of 10, $14.00 !!
These are the ones I used.
Still waiting for my stains, from the UK.
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

Pau
Site Admin
Posts: 6052
Joined: Wed Jan 20, 2010 8:57 am
Location: Valencia, Spain

Re: Hand sectioning; plants

Post by Pau »

Straight scalpel blades are useful for hand cutting, although in most cases too short for use with typical hand microtomes
Swann-Morton #16 may be convenient,
https://www.ebay.co.uk/itm/Swann-Morton ... SwSXFeEO8r

I've used their "Minor Skin Graft Blade No. SG3" with good success with vegetable not too soft tissues
https://www.swann-morton.com/product/33.php
Pau

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

Re: Hand sectioning; plants

Post by NikonUser »

My botanical stains finally arrived (from the UK).
Here are two sections of a lupine stem using the double-blade method; stained with Safranin-O and Fast Green.
Reversed 40mm enlarging lens and Nikon 10x for close up, Nikon D700
Rather thick sections but considering the cost of equipment (= 2 razor blades) it seems a useful technique for casual interest in plant anatomy.
Attachments
lupine 7iii21 whole.jpg
lupine 7iii21 crop.jpg
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

Sumguy01
Posts: 1715
Joined: Mon Jan 28, 2013 11:05 pm
Location: Ketchikan Alaska USA

Re: Hand sectioning; plants

Post by Sumguy01 »

=D> Very nice.
Last one is amazing.
Thanks for sharing.

klevin
Posts: 129
Joined: Fri Dec 14, 2012 5:28 pm
Location: SW New Hampshire, USA

Re: Hand sectioning; plants

Post by klevin »

Tell me more about the staining? Any beginner references?

Great pics!

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

Re: Hand sectioning; plants

Post by NikonUser »

Thanks Sumguy01; it seems that keeping the blades perpendicular to the stem results in a complete section but rather thick. I do sometimes end up with some thin sections but these are never complete 'circles'; so I get maybe 1/2 a section but the inner straight edge is just a few cells thick. Likely cause is the blades cutting at an angle less than 90.
Klevin; simply dissolve minute amounts of stain in water, basic ref:
http://microscopy.berkeley.edu/Resource ... aining.htm
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

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

Re: Hand sectioning; plants

Post by NikonUser »

Just used the 2-blade technique to section a White Pine twig. Difficult, could get only a half section.
The large holes and small ones in the xylem are resin ducts.
The stem as shown is 1.40 mm across the midline.
Blue stain is Fast Green. Red stain is Safranin-0, used as a very dilute solution in water and it still stained too heavily!
Attachments
19iii21 pine.jpg
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

rjlittlefield
Site Admin
Posts: 23561
Joined: Tue Aug 01, 2006 8:34 am
Location: Richland, Washington State, USA
Contact:

Re: Hand sectioning; plants

Post by rjlittlefield »

White Pine twig
Nice image!

But I see two arrows labeled "cambium". Should the one at lower right be "cortex"?

--Rik

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

Re: Hand sectioning; plants

Post by NikonUser »

Thanks Rik.
2 sets of cambium tissue; one gives rise to the xylem and phloem and the other to the outer bark. Damaging the outer bark often results in damaging the outer cambium tissue - often bad news for a tree.
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

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

Re: Hand sectioning; plants

Post by NikonUser »

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!
Attachments
24iii21 carrot sections.jpg
24iii21 carrot section.jpg
24iii21 carrot xylem.jpg
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

Lou Jost
Posts: 5944
Joined: Fri Sep 04, 2015 7:03 am
Location: Ecuador
Contact:

Re: Hand sectioning; plants

Post by Lou Jost »

Those are beautiful carrot cross-sections.

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