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

[Due, Brice]

John, I'm being overly broad, I'm sure, by calling it a "nissl". You list it as
a counterstain for LFB, p124 of your text. Conceptually I always think of LFB+CV
(=KB) as  being LFB plus Nissl counterstain. The "KB" procedure here is exactly
that, an LFB followed by a Nissl. Can a given cv procedure be a counterstain,
but not a nissl when used as a primary stain? Sloppy terminology is very
probably my mistake. My apologies. However your oxalic cv works well for me as a
nissl and is cleaner than the purely regressive one on the books here.

In any case, when I read another reply I realized the poster is working with FS,
so I shouldn't have replied in the first place. Too little sleep, too much
caffeine...

Thanks,
-brice


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-----Original Message-----
From: John A. Kiernan [mailto:jkiernan <@t> uwo.ca]
Sent: Tuesday, October 18, 2005 2:25 PM
To: Due, Brice
Cc: histonet
Subject: Re: [Histonet] RE: Nissl on Thick Paraffin...


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
> 
> "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR
> ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED
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> OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER
> THAN THE INTENDED RECIPIENT IS PROHIBITED.  IF YOU RECEIVE THIS EMAIL IN
ERROR,
> PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER."
> HIPPA Policy, 2003,  Brigham & Women's Hospital, Boston, MA
> 
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