[Histonet] X-gal staining

[Paula Keene Pierce]

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 at 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-request at lists.utsouthwestern.edu <histonet-request at lists.utsouthwestern.edu>

Sent: 08 January 2020 18:00

To: histonet at lists.utsouthwestern.edu <histonet at 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)



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Message: 1

Date: Tue, 7 Jan 2020 18:21:31 +0000

From: "Margaryan, Naira" <naira.margaryan at hsc.wvu.edu>

To: "'histonet-request (histonet at lists.utsouthwestern.edu)'"

        <histonet at lists.utsouthwestern.edu>

Subject: [Histonet] X-gal staining

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        <MN2PR05MB71011189AB120F740D180AB0A53F0 at MN2PR05MB7101.namprd05.prod.outlook.com>

        

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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" <CWaitts at Centrastate.com>

To: "'histonet at lists.utsouthwestern.edu'"

        <histonet at lists.utsouthwestern.edu>

Subject: [Histonet] (no subject)

Message-ID:

        <131bf77daa5d4ba08f9076d9afbf1f65 at exchnode1.local.centrastate.com>

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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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