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PostPosted: Sun Apr 14, 2019 7:12 pm 
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Hi all, a few images of some slides I made yesterday from tissue taken from a nice Carex.pendula AKA 'Pendulous sedge' that is growing in my garden. This is a monocotyledonous plant and has some very nice anatomy that I thought some may enjoy.

These sections were cut at 10µ and 5µ. Generally the thicker sections will show overall morphology and stain more heavily for the same staining protocol, whilst the 5µ (in this case) ones will show more fine detail and stain more lightly - a little counter-intuitive maybe, but more tissue basically means more stain!

Here are a few images, all stained with Safranin and Fast-green.

The whole cross-section from several stitched images through the 'scope...
Image

Some interesting detail of the area around the main-vein, this image is actually from a 5µ section.
Image

This is a region up to the edge of the leaf, with parts labelled...
Image

I particularly like the tiny faces of the vascular bundles.... 10µ really stains brightly,
Image

Same region at 5µ, slightly lighter stain and finer detail...
Image

Close-up of main-vein. The torn-looking hole above the two large 'eyes' (xylem vessels) of the vascular-bundle is where a large xylem vessel has ruptured as the tissue has expanded during rapid growth - commonly seen in mature xylem.
Image

Nice view of a vascular-bundle showing also the fiber bundles above and below the vascular-bundle, thus completing the 'strut' that includes the vascular-bundle and supports the leaf surfaces. Between these 'struts' (see 1st image and others above) mature leaves have air-spaces, slightly less mature leaves may still have parenchyma in this position.
Image

Forgot this image,
Image

That's all I have for now, I only sectioned the leaves and also some of a large 'Day Lily' (Hemerocallis.fluva) in my garden. At this time I'm building a modest collection of slides showing different types of leaf, these and the Lily being monocotyledonous, together with the anatomical/morphological features that adapt them to their particular natural environments.
I should have a few images from the Lily sections soon also.

Hope you like them! :D :D

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John B


Last edited by mrsonchus on Sun Apr 14, 2019 10:38 pm, edited 1 time in total.

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PostPosted: Sun Apr 14, 2019 7:40 pm 
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Beautiful work [as usual], John

Thanks for sharing

MichaelG.

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PostPosted: Sun Apr 14, 2019 10:43 pm 
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Very nice sections and photographs, and the descriptions, explanations and labelling are particularly appreciated.

Thanks for sharing.

Tom W.


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PostPosted: Mon Apr 15, 2019 9:59 am 
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MichaelG & tgss, many thanks chaps, thanks for your kind comments. :)

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PostPosted: Mon Apr 15, 2019 6:01 pm 
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Hi all, another nice section!

This is a section through the lower (near base) zone of a developing shoot of the Carex.pendula plant. The outer three layers are each a fused leaf, typical of monocotyledons and especially grasses, and in this case sedges. Together these fused blades form a tube which supports the later leaves and flower-spikes as they develop. The upped regions of these and other leaves will open out into blades, but form these supporting sheaths at their lower regions.

From the fourth (inclusive) layer the leaves may be seen to be blades, rather than fused all the way round. In the center is a stem, around which a series of younger stems and their accompanying developing floral spikes may be seen to be developing. This arrangement will ultimately result in the long, pendulous flower spikes that give Carex.pendula it's name, and it's typical grass-like form.

Image

A labelled version of the above image....
Image

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PostPosted: Mon Apr 15, 2019 8:12 pm 
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Hi John
Definitely another winner. Excellent material. You mentioned in your initial post the the sections shown were either 5 or 10 microns. Pretty thin for plant sections - what type of microtome are you using?

Regards
Tom W.


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PostPosted: Mon Apr 15, 2019 9:16 pm 
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Perfect, as usual, Mr. John B.!!...

BillT


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PostPosted: Mon Apr 15, 2019 9:30 pm 
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tgss wrote:
Hi John
Definitely another winner. Excellent material. You mentioned in your initial post the the sections shown were either 5 or 10 microns. Pretty thin for plant sections - what type of microtome are you using?

Regards
Tom W.



Hi Tom, I use a Shandon 0325 retracting rotary microtome with 'Feather' disposable blades, grade S35. I routinely section at 10µ for a 'quick look' at tissue. This after a dip in Toluidine-blue while still in wax section form, allows me to make a quick assessment of a section or series of sections for their integrity and to see if I have the desired plane of sectioning and am at the desired 'place in the tissue'.

This is the voracious beast - the 'Mighty Shandon' as you'll see mentioned in many of my posts and threads - I found her unused and in mint condition for a ridiculously fair price about 3 years ago, and haven't looked back!
Attachment:
File comment: The MIGHTY SHANDON!
ws_shandon_on_table_1.jpg
ws_shandon_on_table_1.jpg [ 200.17 KiB | Viewed 160 times ]


For an overall morphological slide I will section at 10 or 12µ, tissue permitting. For finer work such as embryology or cell division I will most often use 5µ, sometimes 3-4µ. I've sectioned pollen grains down to the 1µ limit of the Shandon, but sections that thin are of limited use if cut from paraffin-wax, better to go to plastic for the 'semi-thin' realm and an 'ultra microtome' that has a microscope attached and is for plastic-embedded tissue.

These sections and slides are all from tissue fully processed through the 'paraffin method' and not untreated 'raw' sectioning as by hand or even freezing microtome.

Thanks for looking. :)

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Last edited by mrsonchus on Mon Apr 15, 2019 9:36 pm, edited 2 times in total.

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PostPosted: Mon Apr 15, 2019 9:31 pm 
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billbillt wrote:
Perfect, as usual, Mr. John B.!!...

BillT


Hi Bill old friend, many thanks for joining my adventure! Pleased you like the images. :D :)

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PostPosted: Mon Apr 15, 2019 9:34 pm 
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You, kind Sir, could publish a textbook... You have a natural talent for botany..

BillT


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PostPosted: Mon Apr 15, 2019 10:24 pm 
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mrsonchus wrote:
Hi Tom, I use a Shandon 0325 retracting rotary microtome with 'Feather' disposable blades, grade S35. I routinely section at 10µ for a 'quick look' at tissue. This after a dip ...

Ahh! Now I understand. 10µ for a 'quick look' eh! Sure, why not :roll: Lovely equipment and lovely work John. I obviously need to have a close look at the rest of your posts, being a newcomer on the forum.
Thanks again.

Tom W.


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PostPosted: Tue Apr 16, 2019 3:18 pm 
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MichaelG. wrote:
Beautiful work [as usual], John

Thanks for sharing

MichaelG.
Like!!

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