[Histonet] RE: Nissl on Thick Paraffin...
BDUE <@t> PARTNERS.ORG
Tue Oct 18 11:31:52 CDT 2005
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.
Brigham & Women's Hospital
"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
MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING
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
More information about the Histonet