[Histonet] RE: Nissl on Thick Paraffin...

[John A. Kiernan]

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
   kiernan[AT]uwo.ca
   http://publish.uwo.ca/~jkiernan/
   http://instruct.uwo.ca/anatomy/530/index.htm
_______________________________
"Due, Brice" wrote:
> 
> Hello Nissl, I deleted your original post by accident & histosearch.com appears
> down...
> 
> 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!
> -brice
> Neuropathology Lab
> Brigham & Women's Hospital
> Boston
> 
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