I have done paraffin processing and sectioning of X Gal stained eyes and embryo
heads.
The staining is done for the blue color first, post fixed, and then
processed.Xylene should be avoided, as it is what removes the X Gal target, and
a xylene substitute used such as StatLab's XS3.
Deparaffinization and dehydration for coverslipping should also be done with
the xylene substitute.
I counterstain with Nuclear Fast Red.
Paula Keene Pierce, BS, HTL(ASCP)HTPresidentExcalibur Pathology, Inc.5830 N
Blue Lake DriveNorman, OK 73069PH 405-759-3953http://www.excaliburpathology.com
A sharp knife is nothing without a sharp eye. - Klingon Proverb
On Saturday, January 11, 2020, 01:22:43 PM CST, Hobbs, Carl via Histonet
<histonet@lists.utsouthwestern.edu> wrote:
Hi
No replies so far so.....my pennyworth.
Imho ...no
beta Gal enzyme is inactivated by std Pwax processing.
So, when you apply the X-gal substrate and chromogen solution to Pwax sections,
it will not give you a positive blue final reaction product
If you want to detect beta Gal in FFPWS and visualise with DAB, you will have
to use an anti beta Gal antibody.
Finding an anti beta Gal enzyme antibody, that works in FFPWS +/- AR is not so
easy.
Good luck
Carl
Carl Hobbs FIBMS
Histology and Imaging Manager
Wolfson CARD
Guys Campus, London Bridge
Kings College London
London
SE1 1UL
020 7848 6813
From: histonet-requ...@lists.utsouthwestern.edu
<histonet-requ...@lists.utsouthwestern.edu>
Sent: 08 January 2020 18:00
To: histonet@lists.utsouthwestern.edu <histonet@lists.utsouthwestern.edu>
Subject: Histonet Digest, Vol 194, Issue 4
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Today's Topics:
1. X-gal staining (Margaryan, Naira)
2. (no subject) (Waitts, Celeste)
----------------------------------------------------------------------
Message: 1
Date: Tue, 7 Jan 2020 18:21:31 +0000
From: "Margaryan, Naira" <naira.margar...@hsc.wvu.edu>
To: "'histonet-request (histonet@lists.utsouthwestern.edu)'"
<histonet@lists.utsouthwestern.edu>
Subject: [Histonet] X-gal staining
Message-ID:
<mn2pr05mb71011189ab120f740d180ab0a5...@mn2pr05mb7101.namprd05.prod.outlook.com>
Content-Type: text/plain; charset="us-ascii"
Hello histonetters,
I have request from a scientist to perform X-gal staining on the formalin fixed
paraffin-embedded kidney.
I was wondering if there is any possibility to do this staining on FFPE tissue
as well as if there any possibility to perform a peroxidase DAB staining for
the X-gal? If so, may I ask you for a detailed protocol to perform this
staining.
Any input and explanation why it is impossible is appreciated.
Thanks in advance,
Naira
------------------------------
Message: 2
Date: Wed, 8 Jan 2020 16:46:18 +0000
From: "Waitts, Celeste" <cwai...@centrastate.com>
To: "'histonet@lists.utsouthwestern.edu'"
<histonet@lists.utsouthwestern.edu>
Subject: [Histonet] (no subject)
Message-ID:
<131bf77daa5d4ba08f9076d9afbf1...@exchnode1.local.centrastate.com>
Content-Type: text/plain; charset="us-ascii"
Question VIP6 SHORT BIOPSY RUN USING CLEARITE
ANYTHING
Celeste Waitts
Histology Supervisor
Centrastate Medical Center
Freehold, NJ 07728
(732-303-5071)
------------------------------
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