Hand section stain colour-fest
Hand section stain colour-fest
Hi all, I'm looking at the pathways taken by vasculature within the stems and organs of some flowering plants, dicotyledonous at that, at the moment. It's a fascinating and very complicated area and I've been having a 'quick peek' in the form of some quickly made hand sections from a shrub that is growing on my drive and has matured into a fine semi-woody mature shrub over the last few years.
Anyway, it's common name is 'Canadian honeysuckle' as far as I know - and it seems a good candidate for some sections and slides for this investigation-to-come. This quick look at the tissues is my usual method before choosing exactly which tissues etc to take as specimens for a new planned set of slides, made usually to aid study of whatever I'm investigating at any time, and at this time it's the vascular (i.e. phloem & xylem).... tissues in their differentiation from the very first primary meristem (producer of new 'blank' cells), the secondary meristem, their differentiation, growth and maturity. It's a very complex arrangement within the plant to run supplies to leaves, branches, root, growing tips, floral and reproductive parts etc - the list is very long indeed!
I thought I'd take a few cross-sections (TS) from the stem and try a few (aqueous) stains on them just to see how the stains reacted with the living tissues. The results are very colourful and I thought you may enjoy a little look at their beautiful shades as they stained the living, hand-sectioned stem sections. I used 4 stains, Toluidine-blue, Methylene-blue, Safranin-O and basic Fuchsin.
These sections are very thick compared with the paraffin sections of course, but are really quite a treat if you enjoy colours!
Here's a piece of the stem cut lengthwise at a leaf-node, a point where the vasculature must branch into the leaf and the often present axial-bud between it's point of emergence from the stem and the stem - the area that I'm interested in....
and,
hmmm.... quite a choice of stains..... I chose Toluidine-blue, Methylene-blue, basic Fuchsin and Safranin...
Here are a couple of sections in each of the four very weakly-mixed aqueous stains for about an hour,
A quick rinse to remove any excess stain gives me these, look OK so far,
Here's a basic-Fuchsin stained section, a very colourful start!
The Toluidine-blue,
and
Methylene-blue,
My favourite here though is the beautiful Safranin-O,
and,
A side-by side view,
and,
I just love the vivid colours, hope you like this quick sideways foray taken during my tissue selection first-steps....
I'll be posting a thread re my pursuit of the vascular pathways or 'traces' as I start that adventure, but it may be a while - I've the sliding-microtome to come into the lab hopefully next week and would like to post some demos of my learning-curve with it as I start off!
p.s. oops Just realized, the basic-Fuchsin was a 50% alcohol mix, probably explains the rather atypical purple-ish tones of the stain... That also explains the turbulence in the staining pot as I added a few drops to the sections in water!
Anyway, it's common name is 'Canadian honeysuckle' as far as I know - and it seems a good candidate for some sections and slides for this investigation-to-come. This quick look at the tissues is my usual method before choosing exactly which tissues etc to take as specimens for a new planned set of slides, made usually to aid study of whatever I'm investigating at any time, and at this time it's the vascular (i.e. phloem & xylem).... tissues in their differentiation from the very first primary meristem (producer of new 'blank' cells), the secondary meristem, their differentiation, growth and maturity. It's a very complex arrangement within the plant to run supplies to leaves, branches, root, growing tips, floral and reproductive parts etc - the list is very long indeed!
I thought I'd take a few cross-sections (TS) from the stem and try a few (aqueous) stains on them just to see how the stains reacted with the living tissues. The results are very colourful and I thought you may enjoy a little look at their beautiful shades as they stained the living, hand-sectioned stem sections. I used 4 stains, Toluidine-blue, Methylene-blue, Safranin-O and basic Fuchsin.
These sections are very thick compared with the paraffin sections of course, but are really quite a treat if you enjoy colours!
Here's a piece of the stem cut lengthwise at a leaf-node, a point where the vasculature must branch into the leaf and the often present axial-bud between it's point of emergence from the stem and the stem - the area that I'm interested in....
and,
hmmm.... quite a choice of stains..... I chose Toluidine-blue, Methylene-blue, basic Fuchsin and Safranin...
Here are a couple of sections in each of the four very weakly-mixed aqueous stains for about an hour,
A quick rinse to remove any excess stain gives me these, look OK so far,
Here's a basic-Fuchsin stained section, a very colourful start!
The Toluidine-blue,
and
Methylene-blue,
My favourite here though is the beautiful Safranin-O,
and,
A side-by side view,
and,
I just love the vivid colours, hope you like this quick sideways foray taken during my tissue selection first-steps....
I'll be posting a thread re my pursuit of the vascular pathways or 'traces' as I start that adventure, but it may be a while - I've the sliding-microtome to come into the lab hopefully next week and would like to post some demos of my learning-curve with it as I start off!
p.s. oops Just realized, the basic-Fuchsin was a 50% alcohol mix, probably explains the rather atypical purple-ish tones of the stain... That also explains the turbulence in the staining pot as I added a few drops to the sections in water!
John B
Re: Hand section stain colour-fest
beautiful. Thanks for the lesson.
Re: Hand section stain colour-fest
Very interesting and nice work! Thank you for sharing!
Re: Hand section stain colour-fest
John, they are all just great. Now you know why I like to do live sections. Reveals lots of detail and also the beauty of the cells.
Looking forward to more but what is 'Canadian honeysuckle' doing in the UK?
JimT
Looking forward to more but what is 'Canadian honeysuckle' doing in the UK?
JimT
Re: Hand section stain colour-fest
Hi John B.,
Thanks for sharing more of your wonderful and interesting work!..
BillT
Thanks for sharing more of your wonderful and interesting work!..
BillT
Re: Hand section stain colour-fest
Great post. Quite beautiful colours. Agree, that Safranin is really striking.
Zeiss Standard WL (somewhat fashion challenged) & Wild M8
Olympus E-P2 (Micro Four Thirds Camera)
Olympus E-P2 (Micro Four Thirds Camera)
Re: Hand section stain colour-fest
Thanks for sharing! A lot to learn! Very beautiful images in my eyes!
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Olympus BX51 | Olympus CX23 | Olympus SZ40 | Carl ZEISS EVO LS 10 Lab6 | Carl Zeiss Jena Sedival
Re: Hand section stain colour-fest
These are wondeful. I also think the colours of the cells and the saffarinin are amazing.
You are a master craftsman John and its lovely to see your results.
You are a master craftsman John and its lovely to see your results.