Hand sectioning; plants
Moderators: rjlittlefield, ChrisR, Chris S., Pau
Hand sectioning; plants
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 . 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
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 . 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
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
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
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- Joined: Tue Aug 01, 2006 8:34 am
- Location: Richland, Washington State, USA
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Re: Hand sectioning; plants
Thanks for the post.
For future reference, here are direct links to the article:
http://www.microscopy-uk.org.uk/mag/artoct20/ms-hand-sectioning-plants.html
http://www.microscopy-uk.org.uk/mag/artoct20/ms-hand-sectioning-plants.pdf
http://www.microscopy-uk.org.uk/mag/artoct20/ms-hand-sectioning-plants/index.html?page=1
--Rik
For future reference, here are direct links to the article:
http://www.microscopy-uk.org.uk/mag/artoct20/ms-hand-sectioning-plants.html
http://www.microscopy-uk.org.uk/mag/artoct20/ms-hand-sectioning-plants.pdf
http://www.microscopy-uk.org.uk/mag/artoct20/ms-hand-sectioning-plants/index.html?page=1
--Rik
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Re: Hand sectioning; plants
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
http://www.microscopy-uk.org.uk/mag/artapr04/wdslicera.html
- MarkSturtevant
- Posts: 1947
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Re: Hand sectioning; plants
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
Dept. of Still Waters
Re: Hand sectioning; plants
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.
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
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
Re: Hand sectioning; plants
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
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
Re: Hand sectioning; plants
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.
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
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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
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
Re: Hand sectioning; plants
Very nice.
Last one is amazing.
Thanks for sharing.
Last one is amazing.
Thanks for sharing.
Re: Hand sectioning; plants
Tell me more about the staining? Any beginner references?
Great pics!
Great pics!
Re: Hand sectioning; plants
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
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
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
Re: Hand sectioning; plants
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!
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
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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
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: 23608
- Joined: Tue Aug 01, 2006 8:34 am
- Location: Richland, Washington State, USA
- Contact:
Re: Hand sectioning; plants
Nice image!White Pine twig
But I see two arrows labeled "cambium". Should the one at lower right be "cortex"?
--Rik
Re: Hand sectioning; plants
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.
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
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
Re: Hand sectioning; plants
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!
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
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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
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
Re: Hand sectioning; plants
Those are beautiful carrot cross-sections.