Kalanchoe

[bernhardinho]

Hi friends

I tried a thin section of a Kalanchoe leaf stem. This is what it looks like, dyed with alciangreen, safranin and fuchsin

100x




Cheers Bernhard

[gpmatthews]

Oh wow! Nice image, glorious colours

[lauriek]

Spectacular shot!

Would this actually show up without the dye? Presumably it would look the same but lack colour, or would it lack definition as well?

[discomorphella]

Hi Bernhard--

Superb staining job. What technic are you using with your triple stain combination? I mostly work on animal tissue, or semiconductors (its a strange life), and the only non-fluorescent plant histological staining I've done used Safranin O / Fast Green FCF (Johansen's formula), which gives similar results. I think I prefer the cell wall staining with the Alcian Green, can you provide a recipe or a reference for the stains?

Thanks,

David

[bernhardinho]

Thanks guys

to answer some questions:

let me remind you that staining in plant sections is being done for analytical reasons basically. We amateurs tend to emphasize the aesthetic aspects of course (and so do I!!) . One of the oldest staining in plant anatomy is Astrablue/Safranin with A. staining (and thus showing) the unlignified materials (cellulose etc.) and S. going for the lignified cells (wood). Now there have been several modifications over the years, the most prominent the one by Etzold using Astrablue/Safranin/Fuchsin. I have replaced the Astrablue by Alcian Green following a sort of new fashion. Whereas the other two dyes were purchased as ready to use solutions by Euromex, the Alcian Green (which is ridiculously expensive) has been provided in a 1 gramm portion of powder by a generous friend. I dissolved it in water.

Now the actual procedure is fairly simple, because the dyes can be applied simultanously!! So after cutting (using a barber's knife and a hand microtome) I put the slices in a bleeching solution (Eau de Javelle) to remove the cell contents (which for impatience reasons wasn't all that successfull here ;-)) Then it needs rinsing in demineralized water and then the stains can be added. Let it soak for three or four minutes, then rinse with water again to remove excess stain. The thin sections will now show an overstaining and have to be differentiated with Ethanol 70%. Normally the safranin covers the blue or green and the time of differentiation depends on when you think (or see under the mic) the color balance is right. In this case I didn't mount the cuts permanently, but put them in glycerol only.


Hope that helps!!


Bernhard

[Perl]

Very Nice Picture

Best Regards
Pär

[Jbailey]

Excellent photo, Bernhard. You put a lot of work into the specimen's preparation. It really paid off, though.

Jim