A Touch of Colour

[mrsonchus]

Hi all, I've been working on stains quite a bit lately, particularly the use of two stains together, a 'main' stain that for example will stain vascular-bundles and a secondary or 'counter' stain that will stain cytoplasm and non-lignified tissues with a contrasting and ideally complementary colour. A classic combination that I've been practicing is that of Safranin with Fast-green.
Also of course there are the stains that do this on their own by virtue of their 'metachromatic' properties, e.g. Toluidine-blue, methylene-blue, Safranin...

Anyway, I've a few nice pictures that I thought I'd put up that folk may like, the vascular-bundles and the teeth along a seed's ribs are some of my favourites;

This is a restrained Safranin/Fast-green stain + counter-stain combination that differentiates nicely this TS of the flower-head and tiny florets therein of a Sonchus, much like a dandelion to look at;

ws_700x525_edited_flower_head_stitched.jpg (108.07 KiB) Viewed 4458 times

The green isn't very vivid but the overall desired effect, that of contrasting the red-pink of the seed food-store tissue from the background with Fast-green has worked pretty well, also I like the fine detail this combination is helping to show..

Here's a similar section only stained with Safranin for comparison..

ws_700x525_slide3_Safranin_floret_TS-0020.jpg (100.96 KiB) Viewed 4458 times

Saranin is a lovely stain and is capable of metachromasia also.

and a Methylene-blue version, a good fine-detail and high-contrast stain in my experience, often underrated?

ws_700x525_m_blue_floret_TS-0002.jpg (101.15 KiB) Viewed 4458 times

Methylene-blue can be surprisingly difficult to use sometimes but it's definitely one of my 'top-5' I think.

I like this section and staining, it's a TS of the base of a seed, showing the branching of vasculature coming into the seed-case from the 'parent plant', where it is routed through the seed-case, not the ovary and ovule within, and onwards (upwards) to supply the flower-parts that sit atop the seed in the Sonchus plant. The differentiation given here my the metachromasia of the superb Toluidine-blue stain is really quite striking,

ws_700x525_labelled_cypsela_TBO_acid_rinse_1.jpg (109.54 KiB) Viewed 4458 times

Closer-in and slightly lower across the seed than the above section the mass of vasculature is picked out pretty clearly in blue by the Toluidine-blue stain,

ws_cypsela_TBO_no_acid-0011.jpg (385.05 KiB) Viewed 4458 times

Some lovely colours for sure! Hope you like them as much as I do folks.

[mrsonchus]

A few more, these are LS of the same seeds, showing good details of the teeth that run along the longitudinal ridges of the seed of Sonchus.oleraceus ('Smooth sow-thistle') feature in so many of my posts..

ws_700x525_cypsela_teeth.jpg (51.79 KiB) Viewed 4453 times

and showing the 'core' or the teeth,

ws_700x525_cyp_teeth_pan_1.jpg (44.62 KiB) Viewed 4453 times

Finally I've a couple of my favourite vascular-bundle stains, both have very nice differentiation between the lignified vessels and the non-lignified accompanying cells,

This is the Toluidine-blue version..

ws_lscape_caex_pendula_leaf-(2.jpg (466.71 KiB) Viewed 4453 times

and my favourite is this Safranin + Fast-green two-stain version - clearly showing the differentiation of primary and secondary cell-walls of the xylem vessels..

ws_lscape_bkg_rem_ICE_x60_v.jpg (399.61 KiB) Viewed 4453 times

Finally a nice close-up of the (Aloe-leaf tissue) Safranin stained (with Fast-green counter-stain) lignified xylem-vessel's secondary cell-wall in pink, a nice clear image from a dry x60 objective as far as I remember - I think I've posted this image before, but I really like it..

ws_aloe_vasc_bundle.jpg (259.99 KiB) Viewed 4453 times

That's my 5 - just felt like a colourful interlude! Hope you enjoy the colours!

[KurtM]

It'll take a while to properly study all these images, but for right now I gotta say that first picture made me gasp. Now that's what I call art!

[75RR]

Agree, first image is a keeper. Must say mrsonchus, your photography is coming along nicely.

[mrsonchus]

Thanks chaps, can't beat a bit of colour!

[rnabholz]

Wow John, what a collection.

As mentioned, a lot to take in here, but a great demonstration of your commitment to the discipline and generosity in sharing it with all of us.

The complexity of some of these structures is wondrous.

Great stuff

Rod

[mrsonchus]

Wow John, what a collection.

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The complexity of some of these structures is wondrous.

Great stuff

Rod

Thanks Rod
This is what I love about botany, every structure and arrangement that makes up a plant is that way for a reason, and I find the sometimes very subtle and hard-to-find differences between species to be of particular interest and extremely stimulating and satisfying to investigate.
I've a lot of posts as you'll see regarding 'Sonchus' which is a 'weed' genus known as 'Sowthistle' that is similar to say a Dandelion, and my ongoing study of the species taxonomy of Sonchus.oleraceus vs that of Sonchus.asper for starters.

Microscopy and histology fascinates me, as does the identification and study of wildflowers here in general in the UK - grasses are particularly difficult at all levels - I will turn a beady-eye upon them this Summer I think!
I also collect dry specimens in a herbarium-press then mounted-sheets for study, as well as a small 'spirit collection' which can of course always be a good source of material for histology too! I love what I do and this forum is a superb place to learn and share with the very talented and friendly folk here!

Thanks for your kind comment and interest - always cheers me up to know others enjoy some of my posts.

[Crater Eddie]

These are all gorgeous! From a purely artistic point of view, I like the first one best.
CE

[JimT]

Awe inspiring! Glad to see someone else using a dry 60X. And to such great effect.

[mrsonchus]

Awe inspiring! Glad to see someone else using a dry 60X. And to such great effect.

Thanks Jim, pleased you like them!

Yes, I use the dry x60 a lot as it definitely shows a greater resolution than my x40, which is also a beautifully sharp objective, but doesn't quite resolve as small as the x60 I've found. I also use the oil x100 a lot as I honestly find it quite quick & easy to use...
I don't however oil the condenser and won't bother unless I buy a seriously-superior 'scope to the one I have now, and I can't see that happening, no real need or indeed desire to do so (not to mention finances..).

The x60 can suffer 'softness' at times, but it's always due to one factor - lack of sufficient contrast in my slide-mounted tissue. It depends really on the purpose of my investigation at the time with any particular slide. I see this more as I learn to target my work towards specific features that I would like to image. If I need utmost magnification, e.g. x60/x100 I will use a stain and degree of staining that gives me a good high-contrast slide, if I need an overall 'birds-eye' view as in the first picture above I will go for colour compliments such as the Safranin/Fast-green combination. The possibilities are endless and the more proficient I become the more I am able to produce specialized slides, often a couple of dozen different preparations of the same tissue, with the intention of using different slides and objective combinations according to what I would like to examine.

The last 5 images above I think may be x60 images, as I know I didn't use oil on any of them - I'm particularly pleased with the resolution shown in the images of cypsela-teeth, very good fine details in those I think.
A fascinating study with huge possibilities! Absolute luxury!

[mrsonchus]

Oh, perhaps some labels would be interesting. Here's the vascular-bundle with labels for the different tissue-types and their functions. remember I'm not a Botanist so this is merely the best I can do with my very limited knowledge.

ws_vasc_bundle_labelled.jpg (308.8 KiB) Viewed 4398 times

The most interesting zone may be the cambium - a 'cell-production' zone of about 2 layers of cells, essentially dividing undifferentiated 'stem-cells' that also occur in other key areas of a plant such as at the root-tip and shoot-tips, and laterally within stems to facilitate thickening.
In the vascular bundles of a stem the cambium produces cells both centripetally (xylem toward the stem-center) and centrifugally (the phloem away from the center and towards the epidermis), that is to say it produces new cells in both directions, which will 'turn into' xylem or phloem accordingly. Fascinating, especially when secondary-growth gives the rather dramatic 'rays' seen in roots!

Can't resist an old picture from last-year's root adventures!
This is what cambium can do when you get it angry!
A TS of a Sonchus root..

ws_root_vascular.jpg (387.26 KiB) Viewed 4398 times

The more I see, the more fascinating plants become!
The first image is stained with Safranin + Fast-green counterstain, the second is stained with the magically-metachromatic Toluidine-blue.

[Seb28]

Lovely work and pictures.

[zzffnn]

Amazing work and beautiful photos! Educational as well. Thank you!

[gekko]

As usual, painstaking work with excellent results. But the first three images are, in my view, a work of visual art.

[mrsonchus]

Thanks chaps, nice to know you like them - I have great pleasure & fun with them!

p.s. I've some nice results in from the Daffodil batch - I'll post the first proper stained pictures later if I get a chance.