[Histonet] RE: Nissl on Thick Paraffin...
John A. Kiernan
jkiernan <@t> uwo.ca
Tue Oct 18 13:25:05 CDT 2005
Cresyl violet with oxalic acid isn't one of mine!
This is the first I've heard about it.
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London, Canada N6A 5C1
"Due, Brice" wrote:
> Hello Nissl, I deleted your original post by accident & histosearch.com appears
> Try John Kiernan's modified cv nissl. It works more progressively than the other
> cv nissls I know, which are purely regressive. 100ml 0.1% cresyl violet spiked
> with the addition of 0.5ml of 1% oxalic acid. See p124 of his book.
> I do nissls on 15-20um paraffin, not the 40um you're talking. Even with John's
> oxalic nissl, I still like to overstain and regress with rosin gum. Gives a
> cleaner background = higher contrast. The rosin is a messy sticky sticky
> procedure, but you can control the differentiation by diluting the rosin.
> After aqueous cv staining of your choice, dehydrate slides in 95% etoh. Use
> 1-10% rosin gum in 95% etoh to differentiate. Make a 10% stock and cut when you
> need finer / slower control. You need a couple changes of rosin soln because it
> gets dirty fast. Dip slides in rosin until stain starts running, then rinse in
> 95% etoh and check on an old scope. Repeat until you get the contrast you want.
> If you take out too much cv, just re-hydrate and start over.
> The main problem with ultra-thick sections is that the front exposed side of the
> tissue will differentiate faster that the back side which is "hidden" against
> the slide. I would try diluting your differentiator (whatever procedure you're
> using) 1:10 and see if you can gain some control that way. Are free-floating
> sections an option? I've never tried that with paraffin embedded stuff. Or maybe
> try a non-cv based progressive nissl. I don't have a good recommendation there.
> Neutral red?
> Good Luck!
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