[Histonet] eosin in 95%

Joe Nocito JNocito <@t> Pathreflab.com
Mon Oct 25 14:46:47 CDT 2004

we put about 30 mLs of our regular staining eosin in our first 100% alcohol.
We used less, but my embedders thought it was too light still.

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

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Joanne
Sent: Monday, October 25, 2004 11:06 AM
To: ctsblack <@t> capeheart.uct.ac.za; histonet <@t> pathology.swmed.edu
Subject: [Histonet] eosin in 95%

Dear list,
Those of you who use eosin in your 95% etoh on the processor-what exactly do
you use? The ready to stain eosin, or more concentrated? I remember someone
writing that they put it in the first 95,correct? We need to stain our tiny
gi bxs to better see them when embedding and cutting. Any tips are

>>> "Melanie Black" <ctsblack <@t> capeheart.uct.ac.za> 10/21/04 02:28AM >>>
>I have also had rod-like crystals in my Alk phos/NBT system. I have
>shown them to the company rep, who has no idea what they might be.
>With so much written about it here...I think DAKO should sit up and
>take a look.

Melanie Black

>Thanks all for the cat scratch advice. I am trying it with HRP- AEC.
>Will report result.
>>>>  "C.M. van der Loos" <c.m.vanderloos <@t> amc.uva.nl> 10/17/04 02:20PM >>>
>Dear Joanne,
>Usually, AP staining doesn't show up in a crystal-like precipitate,
>but I have seen it a couple of times. Never found out what was the
>cause of it. Be sure that after your IHC procedure and before the AP
>visualization, the sections are washed at with a Tris-HCl pH8.5
>buffer. Phosphate ions from PBS will inhibit your AP reaction. You
>also may mix the Permanent Red components first and than press
>immediately through a syringe-filter to remove all bigger particles
>in the AP solution.
>Have you viewed your slides under a fluorescence microscope? A small
>amount of Permanent Red reaction product also gives a bright red
>fluorescence signal when using the rhodamine filter pack (green
>Good suggestion by Richard to use a tonsil as a negative control.
>But, how about running the good old control blanc slide without any
>primary antibody?
>Hope this helps.
>Chris van der Loos, PhD
>Dept. of Pathology
>Academical Medical Center
>Amsterdam - The Netherlands
>----- Original Message -----
>>From  "Joanne Mauger" <MAUGER <@t> email.chop.edu>
>Date  Fri, 15 Oct 2004 15:45:05 -0400
>To  histonet <@t> lists.utsouthwestern.edu, jkiernan <@t> uwo.ca
>Subject  Re: [Histonet] IHC withquantum dots
>Hi group,
>Can anyone help me with suggestions-I am working up the monoclonal
>Ab (H2A10) to cat scratch disease. Ab from Novus Biologicus. I have
>a purchased control from Newcomer Supply. I am using permanent red
>chromagen from Dako., with alk phos link also from Dako. I am
>getting a nice specific staining of what looks like specs vs tiny
>rods or spirochetes, but the control tissue does , by Warthin
>Starry, look much more spirochete-ish. Can I trust this result? I
>have also stained several suspected + cases and see the same dust
>like + staining,but I am afraid it is teeny bits of precipitate.
>Does this ring a bell?
>Please help.
>----- Original Message -----
>>From  "Richard Cartun" <Rcartun <@t> harthosp.org>
>Date  Fri, 15 Oct 2004 16:45:57 -0400
>To  <MAUGER <@t> email.chop.edu>,<histonet <@t> lists.utsouthwestern.edu>,
><jkiernan <@t> uwo.ca>
>Subject  Re: [Histonet] IHC withquantum dots
>Have you tried staining a section of tonsil to see if you get the
>same pattern of staining.  I have found "precipitation" to be a
>problem with alkaline phosphatase (PA) detection, although I am by
>no means an expert when it comes to AP IHC.  I use HRP/AEC for most
>of my infectious disease studies including Bartonella henselae.
>Don't forget that you may not see the normal morphology of the
>bacterium with IHC.  You are labeling epitopes on the surface of the
>organism; you are not staining the entire cell wall.  I have found
>B. henselae to be one of the most difficult organisms to label (and
>interpret) with IHC.  Good luck!
>Richard W. Cartun, Ph.D.
>Director, Immunopathology & Histology
>Assistant Director, Anatomic Pathology
>Hartford Hospital
>80 Seymour Street
>Hartford, CT  06102
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu

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