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-----Original Message-----
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Subject: Histonet Digest, Vol 182, Issue 3
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Today's Topics:
1. IHC Negative reagent controls (Cayman Fleck)
2. Re: IHC Negative reagent controls (Tony Henwood (SCHN))
3. AM to statlab and recommendations (Heather Marlatt)
4. Re: IHC Negative reagent controls (John Garratt)
5. IHC-H&E-IHC HELP... (Curt)
----------------------------------------------------------------------
Message: 1
Date: Thu, 3 Jan 2019 02:48:10 +0000
From: Cayman Fleck <caymanfl...@hotmail.com>
To: "histonet@lists.utsouthwestern.edu"
<histonet@lists.utsouthwestern.edu>
Subject: [Histonet] IHC Negative reagent controls
Message-ID:
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com>
Content-Type: text/plain; charset="iso-8859-1"
A question that came up regarding negative reagent controls for
IHC...currently using Ventana i-View. Our regular negative control goes
through the standard antigen retrieval steps, like 99% of our antibodies.
However there are a small number of antibodies that require enzyme as well
(Protease 1). I've seen a number of suggestions regarding this for the
negative reagent control...some say use an additional negative control
protocol that includes the protease, some say to use a single negative
control protocol and just include the harshest cell conditioning that any of
your protocols use (so basically use the cell conditioning + protease
negative control for all antibodies)...i-View is not polymer-based so we
need to continue using negative controls. Any thoughts or advice?
Frank
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------------------------------
Message: 2
Date: Thu, 3 Jan 2019 04:06:57 +0000
From: "Tony Henwood (SCHN)" <tony.henw...@health.nsw.gov.au>
To: Cayman Fleck <caymanfl...@hotmail.com>
Cc: "histonet@lists.utsouthwestern.edu"
<histonet@lists.utsouthwestern.edu>
Subject: Re: [Histonet] IHC Negative reagent controls
Message-ID:
<0e5c3ac509b7424e942160149bb36...@svdcmbx-mex024.nswhealth.net>
Content-Type: text/plain; charset="us-ascii"
Hi Cayman,
Unfortunately, applying HIER to a negative control for an antibody that
requires enzyme retrieval (or no retrieval at all) is not appropriate.
The pre-treatment processes are different and could unmask different
epitopes.
If you are using a negative control then the whole procedure needs to be
same with the exception or replacing the localisation antibody with an
Isotypic antibody solution. (Isotype controls are primary antibodies that
lack specificity to the target, but match the class and type of the primary
antibody used in the application.)
For example, applying citrate or EDTA HIER to sections prior to using the
CD99 antibody (clone 12E7) can reveal perinuclear (golgi-like) staining of
some tumours (eg some colonic carcinomas) but this is not seen if enzyme
retrieval is used.
Hope this is useful
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA)
Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow,
School of Medicine, University of Western Sydney
Tel: 612 9845 3306
Fax: 612 9845 3318
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001,
Westmead NSW 2145, AUSTRALIA
-----Original Message-----
From: Cayman Fleck via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Thursday, 3 January 2019 1:48 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC Negative reagent controls
A question that came up regarding negative reagent controls for
IHC...currently using Ventana i-View. Our regular negative control goes
through the standard antigen retrieval steps, like 99% of our antibodies.
However there are a small number of antibodies that require enzyme as well
(Protease 1). I've seen a number of suggestions regarding this for the
negative reagent control...some say use an additional negative control
protocol that includes the protease, some say to use a single negative
control protocol and just include the harshest cell conditioning that any of
your protocols use (so basically use the cell conditioning + protease
negative control for all antibodies)...i-View is not polymer-based so we
need to continue using negative controls. Any thoughts or advice?
Frank
Sent from Outlook<http://aka.ms/weboutlook>
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------------------------------
Message: 3
Date: Thu, 3 Jan 2019 08:19:15 -0700
From: Heather Marlatt <hmarlat...@gmail.com>
To: "histonet@lists.utsouthwestern.edu"
<histonet@lists.utsouthwestern.edu>
Subject: [Histonet] AM to statlab and recommendations
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Content-Type: text/plain; charset="UTF-8"
Did anyone else use American Mastertech and is now super frustrated with
Stat Lab???
I?ve used AM for years and loved them, great prices and fast shipping, now
cost is through the roof and they?re shipping all my stuff separately
costing a fortune!
Looking for recommendations on xylene alternatives and safe mounting medium.
I?ve been using transcend and cover safe two American Mastertech signature
products but I?m running for my life.
I?ll be contacting a few of the other vendors I use as well but I?d like
some recommendations on xylene alternatives we?ve been xylene free for 4
years and I?m not going back.
Thank you
Heather
------------------------------
Message: 4
Date: Thu, 03 Jan 2019 16:41:03 +0000
From: John Garratt <john.garr...@ciqc.ca>
To: Cayman Fleck <caymanfl...@hotmail.com>
Cc: "histonet@lists.utsouthwestern.edu"
<histonet@lists.utsouthwestern.edu>
Subject: Re: [Histonet] IHC Negative reagent controls
Message-ID:
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CsZYaD2omb18Ivy0R15jWj3aL5Ws1jlg=@ciqc.ca>
Content-Type: text/plain; charset=UTF-8
The article referenced below may be of interest.
Standardization of Negative Controls in Diagnostic Immunohistochemistry:
Recommendations From the International Ad Hoc Expert Panel
https://www.researchgate.net/publication/261516067_Standardization_of_Negati
ve_Controls_in_Diagnostic_Immunohistochemistry_Recommendations_From_the_Inte
rnational_Ad_Hoc_Expert_Panel
John
www.ciqc.ca
??????? Original Message ???????
On Wednesday, January 2, 2019 6:48 PM, Cayman Fleck via Histonet
<histonet@lists.utsouthwestern.edu> wrote:
> A question that came up regarding negative reagent controls for
IHC...currently using Ventana i-View. Our regular negative control goes
through the standard antigen retrieval steps, like 99% of our antibodies.
However there are a small number of antibodies that require enzyme as well
(Protease 1). I've seen a number of suggestions regarding this for the
negative reagent control...some say use an additional negative control
protocol that includes the protease, some say to use a single negative
control protocol and just include the harshest cell conditioning that any of
your protocols use (so basically use the cell conditioning + protease
negative control for all antibodies)...i-View is not polymer-based so we
need to continue using negative controls. Any thoughts or advice?
>
> Frank
>
> Sent from Outlookhttp://aka.ms/weboutlook
>
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 5
Date: Thu, 3 Jan 2019 17:05:21 +0000
From: Curt <c.ta...@pathologyarts.com>
To: "histonet@lists.utsouthwestern.edu"
<histonet@lists.utsouthwestern.edu>
Subject: [Histonet] IHC-H&E-IHC HELP...
Message-ID:
<9c8f910f72893643b3c3793c3d67132b94d03...@pathologyserver.pathologyarts.loca
l>
Content-Type: text/plain; charset="us-ascii"
Need help before I turn a mistake into an irreparable mistake...
We have some unstained slides that were supposed to get stained with H&E but
my guy stained them with IHC. It's complicated, we received slides and a
block, the block was for IHC, the unstained slides were for H&E, he inverted
the process) The point is, now the unstained slides are stained with IHC...
I know we cannot destain the IHC but we can simply run and H&E over them...
the real question I have is subsequent to the H&E... this pathologist
generally likes to see the H&E then order IHC on them based on what he sees
(we only have these few unstained slides, don't have blocks to recut)...
So the question is... if we've already run IHC, then followed that with and
H&E, can we return to run IHC on the slides again? would you want to skip
any pre-treatment, antigen retrieval????
I don't see this working too well myself, if they're already stained with
DAB, that would be present on the second stain...
Thoughts?
Thanks for your help.
Curt
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