A little more 2-stain work

[mrsonchus]

Hi again, I almost forgot to post this bit of staining that I dabbled with over the weekend - it's a bit more practice with the 2-stain process that I started to experiment with in earlier posts, the Safranin-Fast-Green classic combo!

The sections used for these practice-runs were not the best, they were chosen for this reason as during many of my staining adventures quite a few casualties occur and end-up being scraped rather rudely from the slide with a razor-blade .

Here's a little of the combo as I practiced with it;
This is a x4 over-view with the condenser's iris closed to give the fullest contrast to the central Safranin-stained vascular (xylem) tissue and the outer Fast-Green stained phloem and ground tissue... The stain is far too light - I needed to stain for longer or with a greater concentration, but the relative strengths of the 2 stains isn't too bad and has some (limited) use, certainly for learning the process...

ws_700x525_x4_S3_high_contrast.jpg (106.2 KiB) Viewed 2516 times

A closer look reveals very good differentiation between the Safranin and F-Green stained tissue - really shows morphological details quite well for an early experiment;

ws_700x525_x60_S3_1.jpg (85.73 KiB) Viewed 2516 times

ws_700x525_x20_S3_stitch.jpg (71.22 KiB) Viewed 2516 times

Fast-Green rules here!

ws_700x525_x20_S3_stitch-0009.jpg (64.98 KiB) Viewed 2516 times

More differentiation coming through,

ws_700x525_x60_S3_2.jpg (81.77 KiB) Viewed 2516 times

Also tried some of the LS stem-sections from my earlier double-embedding trials, see next post - this one's used-up all 5 images...

[mrsonchus]

This is a 2-stained longitudinal-section of a stem of the same (Sonchus.asper) plant as seen in XS earlier, the vascular-tissue is quite remarkable here and shows (or appears to show) at least 2-types of vascular construction, the pitted vessel-walls seen also in earlier posts, and the discreet (as different from the one-piece spiral) rings arrangement of vessel-wall reinforcement of the xylem vessels.
I've read a little re vasculature and it would seem that the pitted-walls belong to elongated cells called 'tracheids', which are individually very long and tapered at their ends, and the 'less primitive' fully-formed xylem vessels with the ring/spiral wall-structure that are shorter individually but are all joined to form a truly 'tubular' xylem vessel of dead tissue. I think both may be evident here, but I'm not sure - I'm certainly only a (very) amateur botanist! (but I love it! )...
The colours are quite striking also;

ws_700x525_x60_stem_LS_vascular-0010.jpg (48.19 KiB) Viewed 2514 times

and,

ws_700x525_stem_LS-0004.jpg (66.18 KiB) Viewed 2514 times

ws_700x525_stem_LS-0007.jpg (84.59 KiB) Viewed 2514 times

and;

ws_700x525_x4_stem_LS_stitchj-0002.jpg (46.32 KiB) Viewed 2514 times

After some further practice with this and other 2-stain counter-staining techniques I'm going to try (gulp ) to inroduce the very vivid 'Orange G' stain that I recently acquired, for a really kaleidoscopic stain-fest!

Here's a little after mixing and filtering some aqueous working-solution of the Orange-G stain, it's quite a colour!

ws_700x525_DSCN2266.jpg (73.15 KiB) Viewed 2514 times

This staining adventure is fascinating, not to mention hugely enjoyable - the though of producing perfectly-stained & mounted slides make my mouth water!

Back soon with some hopefully better quality results than these somewhat limited first-try efforts! What fun!

[billbillt]

Hi John,

I found your post and photos very enjoyable and as always very pleasing to the eye.... I always look forward to your postings!...

BillT

[vasselle]

Bonjour.
Très bien réaliser et commenter.
Bravo pour votre travail et merci beaucoup pour le partage.
Bonne continuation.
Cordialement seb