RE: [Histonet] RE: Nissl on Thick Paraffin...
|From:||"Due, Brice" (by way of histonet)|
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
(=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
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
so I shouldn't have replied in the first place. Too little sleep, too much
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From: John A. Kiernan [mailto:firstname.lastname@example.org]
Sent: Tuesday, October 18, 2005 2:25 PM
To: Due, Brice
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
"Due, Brice" wrote:
> Hello Nissl, I deleted your original post by accident & histosearch.com
> Try John Kiernan's modified cv nissl. It works more progressively than the
> cv nissls I know, which are purely regressive. 100ml 0.1% cresyl violet
> 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
> 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
> need finer / slower control. You need a couple changes of rosin soln because
> gets dirty fast. Dip slides in rosin until stain starts running, then
> 95% etoh and check on an old scope. Repeat until you get the contrast you
> 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
> tissue will differentiate faster that the back side which is "hidden" against
> the slide. I would try diluting your differentiator (whatever procedure
> 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
> try a non-cv based progressive nissl. I don't have a good recommendation
> Neutral red?
> Good Luck!
> Neuropathology Lab
> Brigham & Women's Hospital
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