[Histonet] Re guinea pig IHC

Sat Feb 2 13:38:05 CST 2019

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

Send Histonet mailing list submissions to
        histonet at lists.utsouthwestern.edu

To subscribe or unsubscribe via the World Wide Web, visit
         https://emea01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.utsouthwestern.edu%2Fmailman%2Flistinfo%2Fhistonet&data=01%7C01%7Ccarl.hobbs%40kcl.ac.uk%7C1546c2da69de40ae400f08d689398cc4%7C8370cf1416f34c16b83c724071654356%7C0&sdata=gEf%2FzAdfyRdZG0CI%2Bty4tXrOxlk5NwyD4O9qxP8DZ0E%3D&reserved=0
or, via email, send a message with subject or body 'help' to
        histonet-request at lists.utsouthwestern.edu

You can reach the person managing the list at
        histonet-owner at lists.utsouthwestern.edu

When replying, please edit your Subject line so it is more specific
than "Re: Contents of Histonet digest..."

Today's Topics:

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

----------------------------------------------------------------------

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.

CONFIDENTIALITY NOTICE:
This e-mail message, including all attachments, is for the sole use of the
intended recipient(s) and may contain confidential and privileged
information. You may NOT use, disclose, copy or disseminate this
information.  If you are not the intended recipient, please contact the
sender by reply e-mail immediately.  Please destroy all copies of the
original message and all attachments. Your cooperation is greatly
appreciated.
Columbus Regional Hospital
2400 East 17th Street
Columbus, Indiana 47201

------------------------------

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>

Content-Type: text/plain; charset="iso-8859-1"

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

*Confidentiality Notice: This message, together with any attachments, is
intended only for the use of the individual or entity to which it is
addressed and may contain confidential or privileged information. If you
think you have received this message in error, please advise the sender and
then delete this message and any attachments immediately.*

------------------------------

Subject: Digest Footer

_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
https://emea01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.utsouthwestern.edu%2Fmailman%2Flistinfo%2Fhistonet&data=01%7C01%7Ccarl.hobbs%40kcl.ac.uk%7C1546c2da69de40ae400f08d689398cc4%7C8370cf1416f34c16b83c724071654356%7C0&sdata=gEf%2FzAdfyRdZG0CI%2Bty4tXrOxlk5NwyD4O9qxP8DZ0E%3D&reserved=0

------------------------------

End of Histonet Digest, Vol 183, Issue 2
****************************************