Rebekah,
How old is your AEC and when do you make it up? I used to get tan-brown
precipitate when I made up the AEC too far in advance.
Also, have you changed lot numbers or vendors recently?

Joe Nocito, BS, HT(ASCP) QIHC
Histology Manager
Pathology Reference Lab
San Antonio, TX


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of SMITH,REBEKAH
FELICIA
Sent: Thursday, July 01, 2004 8:53 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] weird brownish stuff in AEC slides

I have yet another question, guys. I've been asking a lot of
questions about blocking endogenous peroxidase,but now I have a
new situation. I'm still using the ABC peroxidase method staining
sheep placenta (formalin fixed, paraffin embedded) for eNOS and
using AEC as a chromogen and gill's hematoxylin (blued with tap
water and water with 2 or 3 drops of ammonia) as a counterstain.
(btw, still blocking peroxidase with 30% peroxide in methanol,
since my professors are away and haven't been able to order
anything better) I have the lovely red and blue contrast in most
parts of my slides, but I also am getting areas of brownish-black
staining (kinda DAB colored, although I'm not using any DAB). I'm
not exactly sure where the brown comes from this and whether its a
problem with my AEC, my hematoxylin or something else. Anyone have
any ideas? Thanks in advance, y'all have already been really
helpful.

--
SMITH,REBEKAH FELICIA
"You are a child of the universe, no less than the trees and the
stars
You have a right to be here and whether or not it is clear to you,
no doubt the universe is unfolding as it should. Therefore be at
peace with G-d, whatever you conceive Him to be. And whatever your
labors and aspirations,in the noisy confusion of life, keep peace
in your soul.-Max Ehrmann,"Desiderata"


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