[Histonet] Re guinea pig IHC

Sun Feb 3 13:53:57 CST 2019

good advise Carl and you asked some of the things I was wondering.

Patsy Ruegg, HT(ASCP)QIHC
Ruegg IHC Consulting
40864 E Arkansas Ave
Bennett, CO 80102
H 303-644-4538
C 720-281-5406
pruegghm at hotmail.com

________________________________
From: Hobbs, Carl <carl.hobbs at kcl.ac.uk>
Sent: Saturday, February 2, 2019 12:38 PM
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] Re guinea pig IHC

Ms Ruegg, as usual , gives excellent advice: avoid HRP.
Use Alk phos?
Block end. alk phos by using levamisole.

However, Ms Shivers does not state the fixation status of her FS.
Is the Gpig tissue perfused-fixed then frozen or....frozen as unfixed...then fixed?
Also....why 0.3% H2O2?
Use 3%....kill the enzyme, not feed it?
NOT aq for FS
Make up in IMS ( 74OP)
No tissue disruption
However: you state that you get loadsa bubbles...so what? Is your section still attached to your slide?
Can you then carry out successful IF/IHC?
If yes....no problem.
Sure, there's the argument that using a coagulant ( alcohol) in block is a No No
I never had a problem....provided that  the Formalin fixation was sufficient for unfixed crosections ( 15 mins)
I do STILL severely dislike FS ( sure, I spent many years in Diagnostic Histopath doing Operative FS)
 Why not use Pwax sections, Ms Shivers?

Curious-illy

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: 02 February 2019 18:00
To: histonet at lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 183, Issue 2

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Today's Topics:

   1. Re: cytology listserv (Webster, Thomas S.)
   2. Re: guinea pig IHC (Patsy Ruegg)

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Message: 1
Date: Fri, 1 Feb 2019 18:28:30 +0000
From: "Webster, Thomas S." <twebster at CRH.org>
To: "'histonet at lists.utsouthwestern.edu'"
        <histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] cytology listserv
Message-ID: <93fc6a1cc62f41f5ad5890786ab042da at CRH.org>
Content-Type: text/plain; charset="us-ascii"

Haven't seen any cytology listservs except the one for members of the ASC.  There are some cytology facebook pages where you could get questions answered. This Histonet listserv is very informative.

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Message: 2
Date: Fri, 1 Feb 2019 18:39:54 +0000
From: Patsy Ruegg <pruegghm at hotmail.com>
To: Jan Shivers <shive003 at umn.edu>, histonet
        <histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] guinea pig IHC
Message-ID:
        <MWHPR1301MB1886DEBBFD50BEF547312962D8920 at MWHPR1301MB1886.namprd13.prod.outlook.com>

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Especially a very blood tissue like GP spleen.

Patsy Ruegg, HT(ASCP)QIHC
Ruegg IHC Consulting
40864 E Arkansas Ave
Bennett, CO 80102
H 303-644-4538
C 720-281-5406
pruegghm at hotmail.com

________________________________
From: Patsy Ruegg <pruegghm at hotmail.com>
Sent: Thursday, January 31, 2019 11:51 AM
To: Jan Shivers; histonet
Subject: Re: [Histonet] guinea pig IHC

In my experience it is not that GP have a higher peroxidase level, it is frozen sections in general that cannot be blocked with h202, unless they are fixed for a long time in formalin.  What are others experiences with h202 blocking on frozen sections.  I always  used an IHC detection system that did not require h202 blocking for frozen sections.

Patsy Ruegg, HT(ASCP)QIHC
Ruegg IHC Consulting
40864 E Arkansas Ave
Bennett, CO 80102
H 303-644-4538
C 720-281-5406
pruegghm at hotmail.com

________________________________
From: Jan Shivers <shive003 at umn.edu>
Sent: Tuesday, January 29, 2019 12:58 PM
To: histonet
Subject: [Histonet] guinea pig IHC

Has anyone ever performed IHC on frozen sections of guinea pig tissue?  I
am experiencing an enormous amount of bubbling when doing the peroxidase
blocking step, even though I'm only using a 0.3% concentration of H2O2.
And when I say 'enormous', I mean it's like continuous champagne bubbles
rising out of the tissue, even after 20 minutes in the H2O2 solution.

I can't find anything in the literature that mentions guinea pigs having a
higher peroxidase content in their tissues.

Thanks for any help that anyone can provide.

Jan Shivers
Senior Scientist
IHC/Histology Section Manager
Pathology Teaching Program
Veterinary Diagnostic Laboratory
University of Minnesota
1333 Gortner Ave.
St. Paul, MN  55108
612-624-7297
shive003 at umn.edu

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