Re: [Histonet] IHC Validation Question

[Greg Dobbin via Histonet]

Hi Kara,
Use multi-tissue block controls with normal tissues that are treated
exactly the same (pre-analytically speaking) as your routine surgical
specimens. Consult NordiQC.org for recommended normal control tissue for
each IHC marker. The most used multi-tissue control block in my lab has
pieces of tonsil, pancreas, small bowel and liver.

*Greg Dobbin*
1205 Pleasant Grove Rd
Route 220
York,  PE  C0A 1P0
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Re: [Histonet] IHC validation after a new tissue processer is installed

[Greg Dobbin via Histonet]

Hi martha,
Prognostic markers must be re-validated (Eg.s Breast markers and CD117) as
you described.
Every Ab in your menu should be  tested (as you would for a new a new lot)
and do not forget to validate your H&E (with various tissue types) and SS
as well. For the H&Es, if possible do side-by-side comparisons between old
and new processors (the downside of what is otherwise an exciting time!).
Greg

-- 
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE  C0A 1P0


*Everything in moderation...even moderation itself**!*
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Re: [Histonet] IHC validation

[Joe Myers via Histonet]

Kristy:
When validating an IHC procedure, regulatory guidelines like CLIA are not 
concerned so much with the types of specimens upon which the procedure will be 
applied as they are with how appropriately a given procedure detects different 
levels of protein expression, which, in turn, usually correlates to the  
‘degree of disease’.   In practical terms, this means that your lab’s efforts 
to validate should involve the staining of different tissues of the same type 
(i.e. skin), where the expression level ranges from ‘low‘ to ‘high’.  There are 
certainly a great deal more issues involved in procedure validation, but this 
is my attempt to answer your initial question.
Joe Myers, M.S., CT/QIHC(ASCP)

***

Message: 2
Date: Wed, 6 May 2020 05:43:22 -0700
From: Kristy Castillo 
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC validation

Would like to know when validating for IHC for a dermatology lab and just
for CLIA (no CAP), do you just need to show a shave, punch and excision 
lighting up?  Thanks!
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Re: [Histonet] IHC validation

[Echague, Caren Ann - MGH via Histonet]

Where do you purchase your Tisssue Micro arrays? I know they are quite 
expensive.
We are currently running validations and using only a handful of positive 
cases. Most of them don't have ten positive and negative cases but as long as 
you can get a statement from the medical director, this should be CAP compliant.
Cae Aguilar, HTL (ASCP)
Histology Supervisor
7079802801

-Original Message-
From: Allan Wang [mailto:all...@biomax.us] 
Sent: Thursday, March 22, 2018 11:02 AM
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] IHC validation

How are labs validating for rarer biomarkers like ALK, ROS1, or MMR loss?
Following the guideline of 10 positive cases may be difficult.
I've seen other companies with control slides just with a few engineered cell 
lines as positive and negative controls. Is that enough for validation alone?

We are creating TMAs for validation of ALK and MMR loss (MLH1, PMS2, MSH6,
MSH2) so I am interested in knowing how many cases are desired.

Allan

Allan Wang
Lab Manager
US Biomax

On Tue, Mar 20, 2018 at 1:54 PM, Terri Braud via Histonet < 
histonet@lists.utsouthwestern.edu> wrote:

> Just another note:  You can order unstained tissue microarrays with 
> the prerequisite number of cases, both positive and negative, and 
> stain your validation all on one slide.  I've done this for years and 
> for 3 different validations of entire IHC platform changes, ranging 
> from 40 to over 100 antibodies each time.  Saves time and money.
>
> Terri L. Braud, HT(ASCP)
> Anatomic Pathology Supervisor
> Laboratory
> Holy Redeemer Hospital
> 1648 Huntingdon Pike
> Meadowbrook, PA 19046
> ph: 215-938-3689
> fax: 215-938-3874
> Care, Comfort, and Heal
>
> Message: 2
> Date: Fri, 16 Mar 2018 06:54:30 -0700
> From: "Paula" 
> Subject: [Histonet] Antibody Validation CLIA
>
> Hello,
> We've been discussing about the quantity of slides to run as a 
> validation for IHC antibodies. We are governed by CLIA, and we would 
> like to know if there is a set number of slides to run for a 
> particular antibody we would like to bring in-house for Validation.  I think 
> CAP requires 20 slides..?
> And so we are asking if there is  a requirement with CLIA to run a 
> certain number of slides, or is it up to us (the laboratory director) 
> to decide how many slides to run for Validation/Verification.
> Thank you in advance
> Paula
>
>
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>

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Re: [Histonet] IHC validation

[Allan Wang via Histonet]

How are labs validating for rarer biomarkers like ALK, ROS1, or MMR loss?
Following the guideline of 10 positive cases may be difficult.
I've seen other companies with control slides just with a few engineered
cell lines as positive and negative controls. Is that enough for validation
alone?

We are creating TMAs for validation of ALK and MMR loss (MLH1, PMS2, MSH6,
MSH2) so I am interested in knowing how many cases are desired.

Allan

Allan Wang
Lab Manager
US Biomax

On Tue, Mar 20, 2018 at 1:54 PM, Terri Braud via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Just another note:  You can order unstained tissue microarrays with the
> prerequisite number of cases, both positive and negative, and stain your
> validation all on one slide.  I've done this for years and for 3 different
> validations of entire IHC platform changes, ranging from 40 to over 100
> antibodies each time.  Saves time and money.
>
> Terri L. Braud, HT(ASCP)
> Anatomic Pathology Supervisor
> Laboratory
> Holy Redeemer Hospital
> 1648 Huntingdon Pike
> Meadowbrook, PA 19046
> ph: 215-938-3689
> fax: 215-938-3874
> Care, Comfort, and Heal
>
> Message: 2
> Date: Fri, 16 Mar 2018 06:54:30 -0700
> From: "Paula" 
> Subject: [Histonet] Antibody Validation CLIA
>
> Hello,
> We've been discussing about the quantity of slides to run as a validation
> for IHC antibodies. We are governed by CLIA, and we would like to know if
> there is a set number of slides to run for a particular antibody we would
> like to bring in-house for Validation.  I think CAP requires 20 slides..?
> And so we are asking if there is  a requirement with CLIA to run a certain
> number of slides, or is it up to us (the laboratory director) to decide how
> many slides to run for Validation/Verification.
> Thank you in advance
> Paula
>
>
> ___
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> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
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Re: [Histonet] IHC validation

[Morken, Timothy via Histonet]

I've used hundreds of TMA's from Pantomics and Biochain with great results. 

https://pantomics.com/

https://www.biochain.com/


Tim Morken
Pathology Site Manager, Parnassus 
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center

-Original Message-
From: Dessoye, Michael via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Thursday, March 22, 2018 9:00 AM
To: Terri Braud; 'histonet@lists.utsouthwestern.edu'
Subject: Re: [Histonet] IHC validation

Can anyone recommend a vendor that they've had good luck with for TMA slides?

Michael J. Dessoye, M.S. | Histology/Toxicology/Special Chemistry Supervisor | 
Commonwealth Health Laboratory Services | mjdess...@commonwealthhealth.net | 
575 N. River Street | Wilkes Barre, PA 18764 | Tel: 570-552-1432 | Fax: 
570-552-1484

-Original Message-
From: Terri Braud [mailto:tbr...@holyredeemer.com] 
Sent: Tuesday, March 20, 2018 1:54 PM
To: 'histonet@lists.utsouthwestern.edu'
Subject: Re: [Histonet] IHC validation

Just another note:  You can order unstained tissue microarrays with the 
prerequisite number of cases, both positive and negative, and stain your 
validation all on one slide.  I've done this for years and for 3 different 
validations of entire IHC platform changes, ranging from 40 to over 100 
antibodies each time.  Saves time and money.

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Laboratory
Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
ph: 215-938-3689
fax: 215-938-3874
Care, Comfort, and Heal

Message: 2
Date: Fri, 16 Mar 2018 06:54:30 -0700
From: "Paula" 
Subject: [Histonet] Antibody Validation CLIA

Hello,
We've been discussing about the quantity of slides to run as a validation for 
IHC antibodies. We are governed by CLIA, and we would like to know if there is 
a set number of slides to run for a particular antibody we would like to bring 
in-house for Validation.  I think CAP requires 20 slides..?
And so we are asking if there is  a requirement with CLIA to run a certain 
number of slides, or is it up to us (the laboratory director) to decide how 
many slides to run for Validation/Verification.
Thank you in advance
Paula




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Re: [Histonet] IHC validation

[Dessoye, Michael via Histonet]

Can anyone recommend a vendor that they've had good luck with for TMA slides?

Michael J. Dessoye, M.S. | Histology/Toxicology/Special Chemistry Supervisor | 
Commonwealth Health Laboratory Services | mjdess...@commonwealthhealth.net | 
575 N. River Street | Wilkes Barre, PA 18764 | Tel: 570-552-1432 | Fax: 
570-552-1484

-Original Message-
From: Terri Braud [mailto:tbr...@holyredeemer.com] 
Sent: Tuesday, March 20, 2018 1:54 PM
To: 'histonet@lists.utsouthwestern.edu'
Subject: Re: [Histonet] IHC validation

Just another note:  You can order unstained tissue microarrays with the 
prerequisite number of cases, both positive and negative, and stain your 
validation all on one slide.  I've done this for years and for 3 different 
validations of entire IHC platform changes, ranging from 40 to over 100 
antibodies each time.  Saves time and money.

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Laboratory
Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
ph: 215-938-3689
fax: 215-938-3874
Care, Comfort, and Heal

Message: 2
Date: Fri, 16 Mar 2018 06:54:30 -0700
From: "Paula" 
Subject: [Histonet] Antibody Validation CLIA

Hello,
We've been discussing about the quantity of slides to run as a validation for 
IHC antibodies. We are governed by CLIA, and we would like to know if there is 
a set number of slides to run for a particular antibody we would like to bring 
in-house for Validation.  I think CAP requires 20 slides..?
And so we are asking if there is  a requirement with CLIA to run a certain 
number of slides, or is it up to us (the laboratory director) to decide how 
many slides to run for Validation/Verification.
Thank you in advance
Paula




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Re: [Histonet] IHC validation

[Ana Maluenda via Histonet]

Hi Terri,

This is actually a very valid note. This has been in my mind for years, but 
never had the chance to put in action in a research environment (I always try 
to bring to the research field the efficiency and money saving strategies we 
use in diagnostics).

Where do you usually order your unstained slides from?

Much appreciated for the advice.

Kind regards,

Ana

Ana Maluenda
Research Assistant
Atherothrombosis and Vascular Biology Laboratory

Baker Heart and Diabetes Institute
75 Commercial Road, Melbourne VIC 3004
P (03) 8532 1359 E ana.malue...@baker.edu.au W www.baker.edu.au


-Original Message-
From: Terri Braud [mailto:tbr...@holyredeemer.com]
Sent: Wednesday, 21 March 2018 4:54 AM
To: 'histonet@lists.utsouthwestern.edu' 
Subject: Re: [Histonet] IHC validation

Just another note:  You can order unstained tissue microarrays with the 
prerequisite number of cases, both positive and negative, and stain your 
validation all on one slide.  I've done this for years and for 3 different 
validations of entire IHC platform changes, ranging from 40 to over 100 
antibodies each time.  Saves time and money.

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Laboratory
Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
ph: 215-938-3689
fax: 215-938-3874
Care, Comfort, and Heal

Message: 2
Date: Fri, 16 Mar 2018 06:54:30 -0700
From: "Paula" 
Subject: [Histonet] Antibody Validation CLIA

Hello,
We've been discussing about the quantity of slides to run as a validation for 
IHC antibodies. We are governed by CLIA, and we would like to know if there is 
a set number of slides to run for a particular antibody we would like to bring 
in-house for Validation.  I think CAP requires 20 slides..?
And so we are asking if there is  a requirement with CLIA to run a certain 
number of slides, or is it up to us (the laboratory director) to decide how 
many slides to run for Validation/Verification.
Thank you in advance
Paula



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Re: [Histonet] IHC validation

[Colleen Forster via Histonet]

I agree Terry,

The TMA slide is a very economical and powerful way to validate with
minimal slides needed.

Colleen Forster

On Tue, Mar 20, 2018 at 12:54 PM, Terri Braud via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Just another note:  You can order unstained tissue microarrays with the
> prerequisite number of cases, both positive and negative, and stain your
> validation all on one slide.  I've done this for years and for 3 different
> validations of entire IHC platform changes, ranging from 40 to over 100
> antibodies each time.  Saves time and money.
>
> Terri L. Braud, HT(ASCP)
> Anatomic Pathology Supervisor
> Laboratory
> Holy Redeemer Hospital
> 1648 Huntingdon Pike
> Meadowbrook, PA 19046
> ph: 215-938-3689
> fax: 215-938-3874
> Care, Comfort, and Heal
>
> Message: 2
> Date: Fri, 16 Mar 2018 06:54:30 -0700
> From: "Paula" 
> Subject: [Histonet] Antibody Validation CLIA
>
> Hello,
> We've been discussing about the quantity of slides to run as a validation
> for IHC antibodies. We are governed by CLIA, and we would like to know if
> there is a set number of slides to run for a particular antibody we would
> like to bring in-house for Validation.  I think CAP requires 20 slides..?
> And so we are asking if there is  a requirement with CLIA to run a certain
> number of slides, or is it up to us (the laboratory director) to decide how
> many slides to run for Validation/Verification.
> Thank you in advance
> Paula
>
>
> ___
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> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
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Re: [Histonet] IHC validation

[Terri Braud via Histonet]

Just another note:  You can order unstained tissue microarrays with the 
prerequisite number of cases, both positive and negative, and stain your 
validation all on one slide.  I've done this for years and for 3 different 
validations of entire IHC platform changes, ranging from 40 to over 100 
antibodies each time.  Saves time and money.

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Laboratory
Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
ph: 215-938-3689
fax: 215-938-3874
Care, Comfort, and Heal

Message: 2
Date: Fri, 16 Mar 2018 06:54:30 -0700
From: "Paula" 
Subject: [Histonet] Antibody Validation CLIA

Hello,
We've been discussing about the quantity of slides to run as a validation for 
IHC antibodies. We are governed by CLIA, and we would like to know if there is 
a set number of slides to run for a particular antibody we would like to bring 
in-house for Validation.  I think CAP requires 20 slides..?
And so we are asking if there is  a requirement with CLIA to run a certain 
number of slides, or is it up to us (the laboratory director) to decide how 
many slides to run for Validation/Verification.
Thank you in advance
Paula


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Re: [Histonet] IHC VALIDATION

[Fulton Regan via Histonet]

Hi Dawn,


I would suggest having a look at this CAP guidance paper, below.  If I can be 
of any assistance, please feel free to contact me. 

1. Patrick, L. F. et al. Principles of analytic validation of 
immunohistochemical assays: Guideline from the College of American Pathologists 
Pathology and Laboratory Quality Center. Arch. Pathol. Lab. Med. 138, 1432–1443 
(2014).


Regan

Regan Fulton, MD/PhD
PhenoPath
551 North 34th Street, Suite 100
Seattle, WA 98103
Phone 206-374-9000   


CEO, Array Science, LLC
475 Gate 5 Road, #102
Sausalito, CA 94965



> On Feb 27, 2018, at 8:34 AM, Olszewski, Dawn via Histonet 
>  wrote:
> 
> Good morning Histo Peeps,
> 
> I was wondering if anyone would be willing to share their protocol on 
> antibody validation for the IHC lab on FFPE tissues.  Do you know if there 
> are specific regulations on validation or does each lab/hospital develop 
> their own methods.  We are no longer under CAP for inspections,  but we are 
> inspected by Joint Commission.
> 
> If you know of specific regulations could you please site them.  We seem to 
> be in disagreement on how this should be done in our lab.
> Thanks for any and all advice in advance.
> 
> Sincerely,
> 
> Dawn Olszewski, HTL(ASCP)QIHC
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RE: [Histonet] IHC Validation:

[Sebree Linda A]

Well said Joelle; we do pretty much the same.

Linda A. Sebree

From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] on behalf of Joelle Weaver 
[joellewea...@hotmail.com]
Sent: Friday, August 29, 2014 4:09 PM
To: Jb; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] IHC Validation:

Yes, ultimately up to lab director/medical director/pathologist as to 
determination of specificity, selectivity, and if you have enough examples, and 
the staining reactivity conforms to the "intended clinical use" during their 
assessment and hopefully approval of your protocol. I have used some TMAs with 
success, especially if you make with your own in-house processed tissues. I try 
to strongly favor using several expression levels of normal and diseased tissue 
whenever possible that reflect what it will be used for in the patient test 
tissues. If single sections, I try use both expected or known negative and 
positive tissue  both normal and diseased , when practical for the first 
validation set when I get past optimization. For small adjustments I may need 
only a few more confirming positives- up to MD in my situation. I  also have 
polymer detection, but I still like some negatives for me. Some people may not 
feel this is necessary, and the pathologist may not need the negatives ( using 
internal controls), but this helps me,  so I do it to feel more confident in my 
results as I present the slides for review. I don't see why you couldn't use 
internal negatives, if you clarify what tissue element acts as the internal 
negative in the tissue type in the validation summary and SOP.  Basically, for 
amount or # to stain, I follow the CAP guidelines  ( newer ones),  for well 
characterized.  For markers with specific guidelines for validation and 
correlation, I follow the CAP guidelines exactly. Setting up the process/SOP s, 
I used the CLSI guidebook on validation of IHC assays. Both resources (CAP & 
CLSI) have been very helpful for me. That is what has been working for me, I 
hope this helps.


Joelle Weaver MAOM, HTL (ASCP) QIHC





> From: craiga...@gmail.com
> Date: Fri, 29 Aug 2014 10:29:15 -0700
> To: Histonet@lists.utsouthwestern.edu
> CC:
> Subject: [Histonet] IHC Validation:
>
> How do most people validate IHC?  I want to create a tissue microarray. I 
> wanted to use an average of 6-8 positive tissues.
>
> Is it necessary to validate using negative tissues when there is always an 
> internal negative control in all tissue sections. Now with new polymer 
> detection systems there is not background, etc.
>
> Is IHC validation ultimately up to the discretion of the laboratory director?
>
> Please advise. Thx-
>
> Sent from my iPhone
> ___
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RE: [Histonet] IHC Validation:

[Sebree Linda A]

I think your last question says it all; it ultimately is up to the director 
although she/he better be able to explain to a CAP inspector why it was done 
the way it was done.  We generally follow the CAP guidelines with 10 - 20 
+/10-20 - cases.  We've also used the internal negative elements as being 
negative as expected in our validation documentation.  In some cases, when + 
cases are rare we've used less than the suggested number just as CAP says is 
acceptable but we've usually been able to come up with enough.  TMAs are a 
great way to go if you have them or can make them; cuts down on labor/reagent 
cost.

Linda A. Sebree

From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] on behalf of Jb 
[craiga...@gmail.com]
Sent: Friday, August 29, 2014 12:29 PM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC Validation:

How do most people validate IHC?  I want to create a tissue microarray. I 
wanted to use an average of 6-8 positive tissues.

Is it necessary to validate using negative tissues when there is always an 
internal negative control in all tissue sections. Now with new polymer 
detection systems there is not background, etc.

Is IHC validation ultimately up to the discretion of the laboratory director?

Please advise. Thx-

Sent from my iPhone
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RE: [Histonet] IHC Validation:

[Joelle Weaver]

Yes, ultimately up to lab director/medical director/pathologist as to 
determination of specificity, selectivity, and if you have enough examples, and 
the staining reactivity conforms to the "intended clinical use" during their 
assessment and hopefully approval of your protocol. I have used some TMAs with 
success, especially if you make with your own in-house processed tissues. I try 
to strongly favor using several expression levels of normal and diseased tissue 
whenever possible that reflect what it will be used for in the patient test 
tissues. If single sections, I try use both expected or known negative and 
positive tissue  both normal and diseased , when practical for the first 
validation set when I get past optimization. For small adjustments I may need 
only a few more confirming positives- up to MD in my situation. I  also have 
polymer detection, but I still like some negatives for me. Some people may not 
feel this is necessary, and the pathologist may not need the negatives ( using 
internal controls), but this helps me,  so I do it to feel more confident in my 
results as I present the slides for review. I don't see why you couldn't use 
internal negatives, if you clarify what tissue element acts as the internal 
negative in the tissue type in the validation summary and SOP.  Basically, for 
amount or # to stain, I follow the CAP guidelines  ( newer ones),  for well 
characterized.  For markers with specific guidelines for validation and 
correlation, I follow the CAP guidelines exactly. Setting up the process/SOP s, 
I used the CLSI guidebook on validation of IHC assays. Both resources (CAP & 
CLSI) have been very helpful for me. That is what has been working for me, I 
hope this helps. 


Joelle Weaver MAOM, HTL (ASCP) QIHC


  

 
> From: craiga...@gmail.com
> Date: Fri, 29 Aug 2014 10:29:15 -0700
> To: Histonet@lists.utsouthwestern.edu
> CC: 
> Subject: [Histonet] IHC Validation:
> 
> How do most people validate IHC?  I want to create a tissue microarray. I 
> wanted to use an average of 6-8 positive tissues. 
> 
> Is it necessary to validate using negative tissues when there is always an 
> internal negative control in all tissue sections. Now with new polymer 
> detection systems there is not background, etc. 
> 
> Is IHC validation ultimately up to the discretion of the laboratory director?
> 
> Please advise. Thx- 
> 
> Sent from my iPhone
> ___
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> Histonet@lists.utsouthwestern.edu
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RE: [Histonet] IHC validation:

[Cartun, Richard]

Many labs have turned to tissue microarrays (TMAs) for validating antibodies 
for IHC testing as a matter of convenience and lower cost.  Ideally, the 
tissues used to make the TMA(s) should be fixed in the formalin that is used in 
your laboratory, as well as processed in your Histology laboratory under 
identical conditions as those used for patient specimens.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & Immunopathology
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596 Office
(860) 545-2204 Fax
richard.car...@hhchealth.org


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jb
Sent: Sunday, August 03, 2014 2:46 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC validation:

Does anyone have experience validating antibodies using tissue microarrays?  Is 
this a preferred method? I was thinking of punching different tissues/cases, 
then embedding them in one block, and then validating the antibody. Please 
advise.

Thank you.

Sent from my iPhone
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Re: [Histonet] IHC validation:

[Sue]

All you would need to do is id the tissue and dx on your paperwork. it should 
not be any different than running each blocks alone. 


susan T. paturzo 
TJU 




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Re: [Histonet] IHC VALIDATION

[Bernadette Del Rosario]

Please share to me as well.thanks..

> On Mar 27, 2014, at 5:06 PM, "Cartun, Richard"  
> wrote:
> 
> I keep an Excel spreadsheet on all my antibodies (and probes).  I list the 
> case number, the tissue tested, the result, "is this the expected result?", 
> diagnosis and/or comments, and the detection system used.  I am a firm 
> believer of maintaining a "prospective" validation meaning I add positive and 
> negative cases to the spreadsheet even after the primary validation is 
> completed.  This allows you to monitor for analytical drift, and also proves 
> useful in identifying cases for educational purposes and for identifying 
> control material.  I will send you (and anyone else who is interested) a copy 
> of the blank template.
> 
> Richard
> 
> Richard W. Cartun, MS, PhD
> Director, Histology & Immunopathology
> Director, Biospecimen Collection Programs
> Assistant Director, Anatomic Pathology
> Hartford Hospital
> 80 Seymour Street
> Hartford, CT  06102
> (860) 972-1596 Office
> (860) 545-2204 Fax
> richard.car...@hhchealth.org
> 
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu 
> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sara 
> Baldwin/mhhcc.org
> Sent: Thursday, March 27, 2014 9:16 AM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] IHC VALIDATION
> 
> Histonetters
> does anyone have a validation form that reflects the new principles of 
> validation out there that would be willing to share???
> 
> Thanks
> Histology/Cytology Supervisor
> S. Kathy Baldwin, SCT (ASCP)
> Memorial Hospital and Health Care Center sbald...@mhhcc.org Ph 812-996-0210, 
> 0216, Fax 812-996-0232, Pager 812-481-0897, Cell 812-887-3357
> 
> "Christ's healing mission of compassion empowers us to be for others through 
> quality and excellence."
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> information. Any unauthorized review, use, disclosure, or distribution is 
> prohibited. If you are not the intended recipient, or an employee or agent 
> responsible for delivering the message to the intended recipient, please 
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RE: [Histonet] IHC VALIDATION

[Cartun, Richard]

I keep an Excel spreadsheet on all my antibodies (and probes).  I list the case 
number, the tissue tested, the result, "is this the expected result?", 
diagnosis and/or comments, and the detection system used.  I am a firm believer 
of maintaining a "prospective" validation meaning I add positive and negative 
cases to the spreadsheet even after the primary validation is completed.  This 
allows you to monitor for analytical drift, and also proves useful in 
identifying cases for educational purposes and for identifying control 
material.  I will send you (and anyone else who is interested) a copy of the 
blank template.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & Immunopathology
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596 Office
(860) 545-2204 Fax
richard.car...@hhchealth.org

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sara 
Baldwin/mhhcc.org
Sent: Thursday, March 27, 2014 9:16 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC VALIDATION

Histonetters
 does anyone have a validation form that reflects the new principles of 
validation out there that would be willing to share???

Thanks
Histology/Cytology Supervisor
S. Kathy Baldwin, SCT (ASCP)
Memorial Hospital and Health Care Center sbald...@mhhcc.org Ph 812-996-0210, 
0216, Fax 812-996-0232, Pager 812-481-0897, Cell 812-887-3357

"Christ's healing mission of compassion empowers us to be for others through 
quality and excellence."
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RE: [Histonet] IHC validation

[Troutman, Kenneth A]

Hi Laurie,

Ideally, you would like to validate both with tissues processed in your 
laboratory (controls and patient tissue).  Purchased TMAs are okay, but they 
should not be the only thing used.

For the instrument validation, I would definitely use tissue that has been 
processed in your lab and run the same stain on each instrument (I think we ran 
serial sections of vimentin) and compare them very closely for any variation 
(use the same antibody and the same detection kit for each instrument).

My recommendation for new antibody validations is to run a set of normal 
tissues and tumor tissues (we have about 15 normal tissues that we harvested 
and created our own sausage block) along with about 20 or so different tumors 
(also created in sausage blocks) to see how they stain under as many 
circumstances as possible (especially in the conditions your pathologists are 
most likely to see).

In addition to that, run a set of known positives and known negatives from your 
cases.  CAP recommends 10 of each, but I believe the final number is at the 
Medical Director's discretion.

You can also run a representative sample of positives and negatives in your lab 
and send another set of the same tissues to another lab that has a validated 
procedure for that antibody and validate it that way.

As far as the detection kit goes, we don't actually "validate" the detection 
kit per se.  We do a lot-to-lot comparison, which involves running tissue from 
the same block (we created special blocks specifically for detection kits) and 
run the same antibody (AE1/AE3 or CD3--something we do a LOT of) to compare 
from lot to lot.

I hope I answered your question.

Regards,

Ashley Troutman BS, HT(ASCP) QIHC
Immunohistochemistry Supervisor
Vanderbilt University Histopathology
1301 Medical Center Drive TVC 4531
Nashville, TN  37232
ashley.trout...@vanderbilt.edu

Message: 21
Date: Fri, 15 Mar 2013 16:51:27 +
From: Laurie Colbert mailto:lcolb...@pathmdlabs.com>>
Subject: [Histonet] IHC validation
To: "Histonet Post 
(histonet@lists.utsouthwestern.edu)"

mailto:histonet@lists.utsouthwestern.edu>>
Message-ID:

<12ECD7346266D74691EC2BFC75285E451CC048EB@BFL323E10.pathmdlabs.local>
Content-Type: text/plain; charset="us-ascii"

If a lab is just starting up IHC for the first time and has to validate both 
the IHC stainer and the AB's/detection kit, does the validation have to be done 
using actual patient cases - or can you use strictly control tissue and 
purchased microarrays?

Laurie Colbert, HT (ASCP)

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RE: [Histonet] IHC validation

[WILLIAM DESALVO]

Laurie, I would use the control slides to validate and set up the antibodies. I 
good secondary check/control would be to send slides to another lab to check 
for correlation and confirmation of your protocol performance. Once you get an 
established process, then you can later check patient samples to the protocol 
and make adjustments, as necessary.

William DeSalvo, BS HTL(ASCP)
Production Manager-Anatomic Pathology
Chair, NSH Quality Management Committee
Owner/Consultant, Collaborative Advantage Consulting

 

> From: lcolb...@pathmdlabs.com
> To: histonet@lists.utsouthwestern.edu
> Date: Fri, 15 Mar 2013 16:51:27 +
> Subject: [Histonet] IHC validation
> 
> If a lab is just starting up IHC for the first time and has to validate both 
> the IHC stainer and the AB's/detection kit, does the validation have to be 
> done using actual patient cases - or can you use strictly control tissue and 
> purchased microarrays?
> 
> Laurie Colbert, HT (ASCP)
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RE: [Histonet] IHC Validation

[burke]

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu]On Behalf Of Dessoye,
Michael J
Sent: Friday, May 04, 2012 7:51 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC Validation


Hello all,
 
A year or so ago, we upgraded from a Benchmark XT to Benchmark Ultra.
For validation, we selected a variety of cases as usual and ran them on
both instruments before we retired the XT.  Now, when we add a brand new
antibody, we again select a variety of cases, and once we're happy with
them on the Ultra we send the same cases to a reference lab for
comparison.  
 
I'm now faced with changing clones for several antibodies.  I expected
to go through pretty much the same validation procedure, but it got me
thinking...the reference lab does not always use the same clone as some
of ours.  I suppose this really wouldn't be a 'true' validation in this
case.
 
Does anyone have any thoughts on this?  The pathologists are perfectly
happy with the staining of the new clone, but the only reference lab I
can use uses a different clone.  Any thoughts on how to perform a good
validation in this case?
 
Michael J. Dessoye, M.S. | Histology Supervisor | Wilkes-Barre General
Hospital | An Affiliate of Commonwealth Health |
mjdess...@commonwealthhealth.net | 575 N. River Street | Wilkes Barre,
PA 18764 | Tel: 570-552-1432 | Fax: 570-552-1526 
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RE: [Histonet] IHC Validation

[Kuhnla, Melissa]

Hi,
Think of yourself as a reference lab as well.  You have a validated
protocol for one clone already.  Validate the new clone against the
current. Your medical director can help you determine how many cases are
sufficient.  Depending on the antibody we validate anywhere from five
cases to twenty.
In my opinion, this in-house validation is the best way.  The only
variable is the clone.  Your processing, staining platform,
detection...all stays the same.
Good luck
melissa

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Dessoye,
Michael J
Sent: Friday, May 04, 2012 8:51 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC Validation

Hello all,
 
A year or so ago, we upgraded from a Benchmark XT to Benchmark Ultra.
For validation, we selected a variety of cases as usual and ran them on
both instruments before we retired the XT.  Now, when we add a brand new
antibody, we again select a variety of cases, and once we're happy with
them on the Ultra we send the same cases to a reference lab for
comparison.  
 
I'm now faced with changing clones for several antibodies.  I expected
to go through pretty much the same validation procedure, but it got me
thinking...the reference lab does not always use the same clone as some
of ours.  I suppose this really wouldn't be a 'true' validation in this
case.
 
Does anyone have any thoughts on this?  The pathologists are perfectly
happy with the staining of the new clone, but the only reference lab I
can use uses a different clone.  Any thoughts on how to perform a good
validation in this case?
 
Michael J. Dessoye, M.S. | Histology Supervisor | Wilkes-Barre General
Hospital | An Affiliate of Commonwealth Health |
mjdess...@commonwealthhealth.net | 575 N. River Street | Wilkes Barre,
PA 18764 | Tel: 570-552-1432 | Fax: 570-552-1526 
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _
_ _

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Re: [Histonet] IHC Validation

[Rene J Buesa]

Hi Michael:
This is what I think, for what it is worth:
1- if you and the reference lab are using different clones, that is not much of 
a validation,
 
2- additionally the validation using another lab poses additional problems: it 
is not only the instrument they are using, but how they use it, what is the 
whole protocol before using the instrument, and how the instrument is 
maintained. At the end of the day, you will compare your results with those of 
the other lab, but qualitatively only.
 
3- now, and this is much more important: if every time you are going to 
implement a new antibody you are going to go through this whole validation 
process it will be very expensive and, as you wrote, your pathologists are 
satisfied.
 
This is how I used to approach this issue: when I changed from manual to the 
DAKO auto stainer, the pathologists were the ones who decided if the results 
were comparable using the same controls I used manually and automated.
 
Each time they wanted a new antibody or clone to be added to our battery of Abs 
I tried several supposedly positive controls and the pathologists either 
accepted or required either increasing or decreasing the Abs dilution to get 
the intensity and reaction pattern they were looking for when comparing with 
the reference they read. They (and I) also compared the results with what the 
literature described as desirable.
 
My validations always rested on the acceptance or rejection from our 
pathologists. They are the ones who are going to use it and no matter who is 
going to "validate" your protocol, the bottom line resides with the likes or 
dislikes of the pathologists. You will never be able to over-rule their 
decision and they are the ones responsible for the whole lab results regarding 
CAP.
 
My advise: rely on your pathologists and never attempt to do a 
costly validation that is not going to be either appreciated or required. You 
can advise your pathologists but they are the "deciders"
René J.
 
 
--- On Fri, 5/4/12, Dessoye, Michael J  wrote:


From: Dessoye, Michael J 
Subject: [Histonet] IHC Validation
To: histonet@lists.utsouthwestern.edu
Date: Friday, May 4, 2012, 8:50 AM


Hello all,

A year or so ago, we upgraded from a Benchmark XT to Benchmark Ultra.
For validation, we selected a variety of cases as usual and ran them on
both instruments before we retired the XT.  Now, when we add a brand new
antibody, we again select a variety of cases, and once we're happy with
them on the Ultra we send the same cases to a reference lab for
comparison.  

I'm now faced with changing clones for several antibodies.  I expected
to go through pretty much the same validation procedure, but it got me
thinking...the reference lab does not always use the same clone as some
of ours.  I suppose this really wouldn't be a 'true' validation in this
case.

Does anyone have any thoughts on this?  The pathologists are perfectly
happy with the staining of the new clone, but the only reference lab I
can use uses a different clone.  Any thoughts on how to perform a good
validation in this case?

Michael J. Dessoye, M.S. | Histology Supervisor | Wilkes-Barre General
Hospital | An Affiliate of Commonwealth Health |
mjdess...@commonwealthhealth.net | 575 N. River Street | Wilkes Barre,
PA 18764 | Tel: 570-552-1432 | Fax: 570-552-1526 
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

This email and any files transmitted with it are confidential and
intended solely for the use of the individual or entity to whom
they are addressed.
If you have received this email in error please notify the
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RE: [Histonet] IHC validation

[Morken, Timothy]

Tim

Use outside controls to validate your own internal controls. Normal control 
tissue is acceptable and is useful because it has consistent expression of the 
antigen. Cancer tissue is quite variable in expression and sometimes the 
expected expression is less than the normal tissue, and sometimes even 
negative. 

As a parallel validation test you can send your unstained slides to a second 
lab that does the test and they send their slides to you. If both results are 
the same you can feel confident in your test.

Tim Morken
Supervisor, Histology, IPOX
UCSF Medical Center
San Francisco, CA, USA

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tim Higgins
Sent: Thursday, September 01, 2011 9:23 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC validation

Hello Histo Friends,

Has anyone had to validate IHC antibodies when you do not have tissue processed 
in house to use for your validation.  I do have control tissue that stains 
properly but it was acquired from outsides sources with verified proper 
predicted staining.

If you have done this type of validation, how did you make the pathologist feel 
comfortable with it?  I understand pathologist all to well and you can't make 
them do anything they do not want to do but I am looking for suggestions that 
might have worked in helping them feel comfortable.  

Any information would be appreciated.

Thanks,

Tim
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Re: [Histonet] IHC validation

[Rene J Buesa]

I had never done that and it would be necessary only if the tissues you are 
using were fixed in a different way to yours.
Tissue fixation is the step that could affect reactivity because if the tissue 
has been fixed in an alcoholic fixative it would not need HIER and it is 
uncertain how this unnecessary step (in that specific case) could affect 
reactivity.
No other step (dehydration and clearing) could really affect the reactivity. 
Even temperatures above 45ºC have been demonstrated not to affect the epitopes.
Xylene do have an extracting effect on epitopes and it would be nice to know if 
the clearing agent was xylene.
Now, if you fix with NBF, dehydrate with ethanol and clear with xylene and the 
tissue you have from outside has been processed with a similar protocol, you do 
not need to validate that tissue. At least is how I see it.
René J.

--- On Thu, 9/1/11, Tim Higgins  wrote:


From: Tim Higgins 
Subject: [Histonet] IHC validation
To: histonet@lists.utsouthwestern.edu
Date: Thursday, September 1, 2011, 12:22 PM


Hello Histo Friends,

Has anyone had to validate IHC antibodies when you do not have tissue processed 
in house to use for your validation.  I do have control tissue that stains 
properly but it was acquired from outsides sources with verified proper 
predicted staining.

If you have done this type of validation, how did you make the pathologist feel 
comfortable with it?  I understand pathologist all to well and you can't make 
them do anything they do not want to do but I am looking for suggestions that 
might have worked in helping them feel comfortable.  

Any information would be appreciated.

Thanks,

Tim
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Re: [Histonet] IHC Validation

[Rene J Buesa]

It will depend in how you do the validation: manually or with a high output 
autostainer.
René J.

--- On Sat, 7/16/11, kira...@sbcglobal.net  wrote:


From: kira...@sbcglobal.net 
Subject: [Histonet] IHC Validation
To: histonet@lists.utsouthwestern.edu
Date: Saturday, July 16, 2011, 8:33 PM


How long it will take to do validation if you have to start with 20 stains ?
Any tips to make it more efficient will be appreciated.
Thank you 
-K
Sent from my Verizon Wireless BlackBerry

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Re: [Histonet] IHC validation

[Richard Cartun]

In my opinion, only if it affects immunoreactivity.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & Immunopathology
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596 Office
(860) 545-2204 Fax


>>> "Ring, Mary L"  4/19/2011 3:04 PM >>>
Does a change in the type of hematoxylin counterstain require any re-validation 
of IHC stains?
Thanks for your help!

Mary Ring, HT, QIHC
Regions Hospital, St Paul, Mn


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Re: [Histonet] IHC validation

[Rene J Buesa]

Not really because the Hematoxylin should "affect" only the nuclei UNLESS it is 
a nuclear antibody, in which case the antigenic reaction could be obscured. 
For nuclear Abs I used to dilute my hematoxylin as much as possible or did not 
use it at all.
René J.

--- On Tue, 4/19/11, Ring, Mary L  wrote:


From: Ring, Mary L 
Subject: [Histonet] IHC validation
To: "histonet@lists.utsouthwestern.edu" 
Date: Tuesday, April 19, 2011, 3:04 PM


Does a change in the type of hematoxylin counterstain require any re-validation 
of IHC stains?
Thanks for your help!

Mary Ring, HT, QIHC
Regions Hospital, St Paul, Mn


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Re: [Histonet] IHC validation

[BSullivan]

Added to this.make sure you keep your work up sheets.


Beatrice Sullivan, HT(A.S.C.P.) HTL , AAS, CLSP(N.C.A.)
AP Supervisor
Shore Memorial Hospital
609-653-3590


Speak only well of people and you need never whisper


   
 Rene J Buesa  
  To
 Sent by:  Histonet
 histonet-bounces@ 
 lists.utsouthwest , Joe Nocito 
 ern.educc
   
   Subject
 02/09/2011 03:26  Re: [Histonet] IHC validation   
 PM
   
   
   
   
   




Joe:
This is what I would do;
1- run 1 control slide per antibody you have in your "arsenal"
2- compare the result with a control slide already in your files.
3- show both slides to the chief pathologist (after all is his/her opinion
the one is going to be asked by CAP)
4- those antibodies whose positive controls reacted substantially different
to those in your files are the ones you have to work with with respect to
concentration or detection method.
5- never overdue it, and avoid excessive costs that usually are never
appreciated.
Rely always in your pathologist's opinion
René J.

--- On Tue, 2/8/11, Joe Nocito  wrote:


From: Joe Nocito 
Subject: [Histonet] IHC validation
To: "Histonet" 
Date: Tuesday, February 8, 2011, 5:43 PM


Greetings Histoland,
I need some help. We are about to switch IHC machines from the
Richard-Allen Axiom to the Ventana Benchmark Ultra. How many slides, per
antibody, do you run for the validation study? We have over 100 primary
antibodies. Normally, when we work up a new antibody, we  start with a
titer. Once that is established, we run 10 cases to check for specificity.
Hopefully we can obtain cases that are really positive, some weakly
positive and some flat out negative. Once that is completed, we run 10
different tissue types to check for any unexpected cross-reactivity. 
The ultra holds 30 slides and we are receiving two machines. If we run 10
slides/antibody, that's going to take a while, not to mention the number of
detection kits that will be used. Do you think 5 slides/antibody is
sufficient? I emailed CAP last week for their take and they never returned
my email (I told my medical director to hold their check for the year and
see how fast they respond to that). Ah oh, don't go down that road Joe,
it's unhealthy. What are your thoughts? Thanks

Joe (JTT)


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RE: [Histonet] IHC validation

[Martha Ward]

What a sensible process!   I agree with your approach. 


Martha Ward, MT (ASCP) QIHC
Assistant Manager
Molecular Diagnostics Lab
Dept. of Pathology
Wake Forest University Baptist Medical Center
Winston-Salem, NC 27157
336-716-2104

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Wednesday, February 09, 2011 3:27 PM
To: Histonet; Joe Nocito
Subject: Re: [Histonet] IHC validation

Joe:
This is what I would do;
1- run 1 control slide per antibody you have in your "arsenal"
2- compare the result with a control slide already in your files.
3- show both slides to the chief pathologist (after all is his/her opinion the 
one is going to be asked by CAP)
4- those antibodies whose positive controls reacted substantially different to 
those in your files are the ones you have to work with with respect to 
concentration or detection method.
5- never overdue it, and avoid excessive costs that usually are never 
appreciated.
Rely always in your pathologist's opinion René J.

--- On Tue, 2/8/11, Joe Nocito  wrote:


From: Joe Nocito 
Subject: [Histonet] IHC validation
To: "Histonet" 
Date: Tuesday, February 8, 2011, 5:43 PM


Greetings Histoland,
I need some help. We are about to switch IHC machines from the Richard-Allen 
Axiom to the Ventana Benchmark Ultra. How many slides, per antibody, do you run 
for the validation study? We have over 100 primary antibodies. Normally, when 
we work up a new antibody, we  start with a titer. Once that is established, we 
run 10 cases to check for specificity. Hopefully we can obtain cases that are 
really positive, some weakly positive and some flat out negative. Once that is 
completed, we run 10 different tissue types to check for any unexpected 
cross-reactivity. 
    The ultra holds 30 slides and we are receiving two machines. If we run 10 
slides/antibody, that's going to take a while, not to mention the number of 
detection kits that will be used. Do you think 5 slides/antibody is sufficient? 
I emailed CAP last week for their take and they never returned my email (I told 
my medical director to hold their check for the year and see how fast they 
respond to that). Ah oh, don't go down that road Joe, it's unhealthy. What are 
your thoughts? Thanks

Joe (JTT)


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RE: [Histonet] IHC validation

[Weems, Joyce]

You know, I can see all this validation if you harvest your own abs and/or use 
concentrates, etc. And also for the FDA approved abs, but I don't see it if we 
use RTU abs that the company has already validated and we are just confirming 
it works in our lab. Seems overkill to me. I like that Rene says it this way! j

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Wednesday, February 09, 2011 15:27
To: Histonet; Joe Nocito
Subject: Re: [Histonet] IHC validation

Joe:
This is what I would do;
1- run 1 control slide per antibody you have in your "arsenal"
2- compare the result with a control slide already in your files.
3- show both slides to the chief pathologist (after all is his/her opinion the 
one is going to be asked by CAP)
4- those antibodies whose positive controls reacted substantially different to 
those in your files are the ones you have to work with with respect to 
concentration or detection method.
5- never overdue it, and avoid excessive costs that usually are never 
appreciated.
Rely always in your pathologist's opinion René J.

--- On Tue, 2/8/11, Joe Nocito  wrote:


From: Joe Nocito 
Subject: [Histonet] IHC validation
To: "Histonet" 
Date: Tuesday, February 8, 2011, 5:43 PM


Greetings Histoland,
I need some help. We are about to switch IHC machines from the Richard-Allen 
Axiom to the Ventana Benchmark Ultra. How many slides, per antibody, do you run 
for the validation study? We have over 100 primary antibodies. Normally, when 
we work up a new antibody, we  start with a titer. Once that is established, we 
run 10 cases to check for specificity. Hopefully we can obtain cases that are 
really positive, some weakly positive and some flat out negative. Once that is 
completed, we run 10 different tissue types to check for any unexpected 
cross-reactivity. 
    The ultra holds 30 slides and we are receiving two machines. If we run 10 
slides/antibody, that's going to take a while, not to mention the number of 
detection kits that will be used. Do you think 5 slides/antibody is sufficient? 
I emailed CAP last week for their take and they never returned my email (I told 
my medical director to hold their check for the year and see how fast they 
respond to that). Ah oh, don't go down that road Joe, it's unhealthy. What are 
your thoughts? Thanks

Joe (JTT)


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Re: [Histonet] IHC validation

[Rene J Buesa]

Joe:
This is what I would do;
1- run 1 control slide per antibody you have in your "arsenal"
2- compare the result with a control slide already in your files.
3- show both slides to the chief pathologist (after all is his/her opinion the 
one is going to be asked by CAP)
4- those antibodies whose positive controls reacted substantially different to 
those in your files are the ones you have to work with with respect to 
concentration or detection method.
5- never overdue it, and avoid excessive costs that usually are never 
appreciated.
Rely always in your pathologist's opinion
René J.

--- On Tue, 2/8/11, Joe Nocito  wrote:


From: Joe Nocito 
Subject: [Histonet] IHC validation
To: "Histonet" 
Date: Tuesday, February 8, 2011, 5:43 PM


Greetings Histoland,
I need some help. We are about to switch IHC machines from the Richard-Allen 
Axiom to the Ventana Benchmark Ultra. How many slides, per antibody, do you run 
for the validation study? We have over 100 primary antibodies. Normally, when 
we work up a new antibody, we  start with a titer. Once that is established, we 
run 10 cases to check for specificity. Hopefully we can obtain cases that are 
really positive, some weakly positive and some flat out negative. Once that is 
completed, we run 10 different tissue types to check for any unexpected 
cross-reactivity. 
    The ultra holds 30 slides and we are receiving two machines. If we run 10 
slides/antibody, that's going to take a while, not to mention the number of 
detection kits that will be used. Do you think 5 slides/antibody is sufficient? 
I emailed CAP last week for their take and they never returned my email (I told 
my medical director to hold their check for the year and see how fast they 
respond to that). Ah oh, don't go down that road Joe, it's unhealthy. What are 
your thoughts? Thanks

Joe (JTT)


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RE:[Histonet] IHC validation

[Troutman, Kenneth A]

I believe the 25 cases are for ER/PR/Her-2 testing.  We have done this recently 
and we ran about 10 for each antibody.  We have over 100 ourselves and it took 
quite a while.  There will be some antibodies that you will be unable to find 
10 slides to test, so do as many (or as few) as your Medical Director is 
comfortable with to validate the stain.

Good luck!

Ashley Troutman BS, HT(ASCP) QIHC
Immunohistochemistry Supervisor
Vanderbilt University Histopathology
1301 Medical Center Drive TVC 4531
Nashville, TN  37232

Message: 14
Date: Tue, 8 Feb 2011 18:12:42 -0500
From: "Weems, Joyce" 
Subject: RE: [Histonet] IHC validation
To: Liz Chlipala , Joe Nocito
  ,  Histonet 
Message-ID:
  <92ad9b20a6c38c4587a9febe3a30e164081e09a...@chexcms10.one.ads.che.org>
Content-Type: text/plain; charset="us-ascii"

But that is for receptors, correct? Do you do that for everything?
Thanks, j

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz Chlipala
Sent: Tuesday, February 08, 2011 17:58
To: Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

Joe

If you are following the recommendations from the CAP paper on IHC 
standardization then it would be 25 tissues (10 strong positive, 10 weak to 
moderate positive and 5 negative).

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 
18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 
www.premierlab.com


Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joe Nocito
Sent: Tuesday, February 08, 2011 3:44 PM
To: Histonet
Subject: [Histonet] IHC validation

Greetings Histoland,
I need some help. We are about to switch IHC machines from the Richard-Allen 
Axiom to the Ventana Benchmark Ultra. How many slides, per antibody, do you run 
for the validation study? We have over 100 primary antibodies. Normally, when 
we work up a new antibody, we  start with a titer. Once that is established, we 
run 10 cases to check for specificity. Hopefully we can obtain cases that are 
really positive, some weakly positive and some flat out negative. Once that is 
completed, we run 10 different tissue types to check for any unexpected 
cross-reactivity.
The ultra holds 30 slides and we are receiving two machines. If we run 10 
slides/antibody, that's going to take a while, not to mention the number of 
detection kits that will be used. Do you think 5 slides/antibody is sufficient? 
I emailed CAP last week for their take and they never returned my email (I told 
my medical director to hold their check for the year and see how fast they 
respond to that). Ah oh, don't go down that road Joe, it's unhealthy. What are 
your thoughts?
Thanks

Joe (JTT)

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RE: [Histonet] IHC validation

[Sally Price]

Liz:
I'd like to know more about these recommendations.  Could you provide a
journal reference to the paper from CAP?
Thx,
Sally

--

Message: 15
Date: Tue, 8 Feb 2011 16:19:53 -0700
From: "Liz Chlipala" 
Subject: RE: [Histonet] IHC validation
To: "Weems, Joyce" , "Joe Nocito"
,"Histonet" histonet@lists.utsouthwestern.edu
Nope that's the recommendation for everything, in the paper they state
additional development is required for prognostic markers. Once you have
validated an antibody it only requires 3 tissues when you get a new lot
number:  1 strong positive, 1 weak to moderate positive and 1 negative.
>From how I read the article its 25 tissues and then 3 tissues, it does
not talk about slides so it is possible to put multiple tissues on one
slide.  Again these are just recommendations; I do not think that there
is a set standard yet.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
-Original Message-
From: Weems, Joyce [mailto:jwe...@sjha.org]
Sent: Tuesday, February 08, 2011 4:13 PM
To: Liz Chlipala; Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

But that is for receptors, correct? Do you do that for everything?
Thanks, j

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz
Chlipala
Sent: Tuesday, February 08, 2011 17:58
To: Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

Joe

If you are following the recommendations from the CAP paper on IHC
standardization then it would be 25 tissues (10 strong positive, 10 weak
to moderate positive and 5 negative).

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joe Nocito
Sent: Tuesday, February 08, 2011 3:44 PM
To: Histonet
Subject: [Histonet] IHC validation

Greetings Histoland,
I need some help. We are about to switch IHC machines from the
Richard-Allen Axiom to the Ventana Benchmark Ultra. How many slides, per
antibody, do you run for the validation study? We have over 100 primary
antibodies. Normally, when we work up a new antibody, we  start with a
titer. Once that is established, we run 10 cases to check for
specificity. Hopefully we can obtain cases that are really positive,
some weakly positive and some flat out negative. Once that is completed,
we run 10 different tissue types to check for any unexpected
cross-reactivity.
The ultra holds 30 slides and we are receiving two machines. If we
run 10 slides/antibody, that's going to take a while, not to mention the
number of detection kits that will be used. Do you think 5
slides/antibody is sufficient? I emailed CAP last week for their take
and they never returned my email (I told my medical director to hold
their check for the year and see how fast they respond to that). Ah oh,
don't go down that road Joe, it's unhealthy. What are your thoughts?
Thanks

Joe (JTT)
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Re: [Histonet] IHC validation

[Joe Nocito]

fortunately, we don't perform prognostic markers. The doctors don't even 
want to venture down that road. When I started working here, that's the 
first ting I mentioned. We can save some much money by doing them in-house. 
That went over like a lead balloon. On a side note, the staff now were 
residents when I retired from active duty. Talk about feeling 
ldd.


Thanks guys for the tips. I will pass them on to the powers at be and let 
them decide.


JTT
- Original Message - 
From: "Morken, Tim" 
To: "'Liz Chlipala'" ; "Weems, Joyce" ; 
"Joe Nocito" ; "Histonet" 


Sent: Tuesday, February 08, 2011 5:37 PM
Subject: RE: [Histonet] IHC validation


When changing instruments you are validating the instrument, not the test. 
For each antibody you just need to run parallel tests in each instrument 
showing equivalence.


However, if you change the instrument and ALSO change the antibody and/or 
detection system, Antigen retrieval, etc, then you need to revalidate the 
test.


For most antibodies (Class I FDA exempt, ancillary tests) that involves 5-10 
positive and negative cases. A small tissue array can suffice. You should do 
reproducibility tests for a few antibodies - 5-10 identical slides on one 
run, 5-10 identical slides on 5-10 runs.


When changing Class II antibodies (ER, PR, Her2) and/or their detection 
systems you will need to run more extensive test. As Liz says, 40 positive, 
40 negative. Again, a tissue array is great for that.


Changing instruments these days is a big deal when a vendor ties you to 
their reagents.


Tim Morken
Supervisor, Histology, IPOX
UCSF Medical Center
San Francisco, CA, USA

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz Chlipala

Sent: Tuesday, February 08, 2011 3:20 PM
To: Weems, Joyce; Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

Nope that's the recommendation for everything, in the paper they state
additional development is required for prognostic markers. Once you have
validated an antibody it only requires 3 tissues when you get a new lot
number:  1 strong positive, 1 weak to moderate positive and 1 negative.

From how I read the article its 25 tissues and then 3 tissues, it does

not talk about slides so it is possible to put multiple tissues on one
slide.  Again these are just recommendations; I do not think that there
is a set standard yet.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949
fax (303) 682-9060
www.premierlab.com


Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: Weems, Joyce [mailto:jwe...@sjha.org]
Sent: Tuesday, February 08, 2011 4:13 PM
To: Liz Chlipala; Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

But that is for receptors, correct? Do you do that for everything?
Thanks, j

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz
Chlipala
Sent: Tuesday, February 08, 2011 17:58
To: Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

Joe

If you are following the recommendations from the CAP paper on IHC
standardization then it would be 25 tissues (10 strong positive, 10 weak
to moderate positive and 5 negative).

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC
PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303)
682-9060 www.premierlab.com


Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joe
Nocito
Sent: Tuesday, February 08, 2011 3:44 PM
To: Histonet
Subject: [Histonet] IHC validation

Greetings Histoland,
I need some help. We are about to switch IHC machines from the
Richard-Allen Axiom to the Ventana Benchmark Ultra. How many slides, per
antibody, do you run for the validation study? We have over 100 primary
antibodies. Normally, when we work up a new antibody, we  start with a
titer. Once that is established, we run 10 cases to check for
specificity. Hopefully we can obtain cases that are really positive,
some weakly positive and some flat out negative. Once that is completed,
we run 10 different tissue types to check for any unexpected
cross-reactivity.
   The ultra holds 30 slides and we are receiving two machines. If we
run 10 slides/antibody, that's going to take a while, not to mention the
number of detection kits that will be used. Do you think 5
slides/antibody is sufficient? I emailed CAP last week for their take
and they never returned my email (I told my medical director to hold
their check for the year and see how fast they respond

RE: [Histonet] IHC validation

[Liz Chlipala]

Thanks Tim

This is a good way of approaching this. What types of antibodies do you
suggest running the reproducibility tests.  For reproducibility I run
about 3 slides in 3 different runs.  I only run reproducibility tests
for antibody cross reactivity studies (just after protocol development)
and for GLP protocol development.  I don't run all of this for my
routine protocol development, but we keep tract of everything we have an
ongoing log for each antibody and lot number and what conditions they
were stained under along with all of the documentation we keep.  

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949 
fax (303) 682-9060
www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: Morken, Tim [mailto:timothy.mor...@ucsfmedctr.org] 
Sent: Tuesday, February 08, 2011 4:38 PM
To: Liz Chlipala; Weems, Joyce; Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

When changing instruments you are validating the instrument, not the
test. For each antibody you just need to run parallel tests in each
instrument showing equivalence. 

However, if you change the instrument and ALSO change the antibody
and/or detection system, Antigen retrieval, etc, then you need to
revalidate the test.

For most antibodies (Class I FDA exempt, ancillary tests) that involves
5-10 positive and negative cases. A small tissue array can suffice. You
should do reproducibility tests for a few antibodies - 5-10 identical
slides on one run, 5-10 identical slides on 5-10 runs.

When changing Class II antibodies (ER, PR, Her2) and/or their detection
systems you will need to run more extensive test. As Liz says, 40
positive, 40 negative. Again, a tissue array is great for that. 

Changing instruments these days is a big deal when a vendor ties you to
their reagents. 

Tim Morken
Supervisor, Histology, IPOX
UCSF Medical Center
San Francisco, CA, USA

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz
Chlipala
Sent: Tuesday, February 08, 2011 3:20 PM
To: Weems, Joyce; Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

Nope that's the recommendation for everything, in the paper they state
additional development is required for prognostic markers. Once you have
validated an antibody it only requires 3 tissues when you get a new lot
number:  1 strong positive, 1 weak to moderate positive and 1 negative.
>From how I read the article its 25 tissues and then 3 tissues, it does
not talk about slides so it is possible to put multiple tissues on one
slide.  Again these are just recommendations; I do not think that there
is a set standard yet.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949 
fax (303) 682-9060
www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: Weems, Joyce [mailto:jwe...@sjha.org] 
Sent: Tuesday, February 08, 2011 4:13 PM
To: Liz Chlipala; Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

But that is for receptors, correct? Do you do that for everything? 
Thanks, j 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz
Chlipala
Sent: Tuesday, February 08, 2011 17:58
To: Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

Joe

If you are following the recommendations from the CAP paper on IHC
standardization then it would be 25 tissues (10 strong positive, 10 weak
to moderate positive and 5 negative). 

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC
PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303)
682-9060 www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joe
Nocito
Sent: Tuesday, February 08, 2011 3:44 PM
To: Histonet
Subject: [Histonet] IHC validation

Greetings Histoland,
I need some help. We are about to switch IHC machines from the
Richard-Allen Axiom to the Ventana Benchmark Ultra. How many slides, per
antibody, do you run for the validation study? We have over 100 primary
antibodies. Normally, when we work up a new antibody, we  start with a
titer. Once that is established, we run 10 cases to check for
specificity. Hopefully we can obtain cases that are really positive,
some weakly positive and some flat out negative. Once that is completed,
we run 10 different tissue types to check for any unexpected
cross-reactivity. 
The ultra holds 30 slides and we are receiving two machines. If we
run 10 slides/antibody, that's going to take a while, not to 

RE: [Histonet] IHC validation

[Morken, Tim]

When changing instruments you are validating the instrument, not the test. For 
each antibody you just need to run parallel tests in each instrument showing 
equivalence. 

However, if you change the instrument and ALSO change the antibody and/or 
detection system, Antigen retrieval, etc, then you need to revalidate the test.

For most antibodies (Class I FDA exempt, ancillary tests) that involves 5-10 
positive and negative cases. A small tissue array can suffice. You should do 
reproducibility tests for a few antibodies - 5-10 identical slides on one run, 
5-10 identical slides on 5-10 runs.

When changing Class II antibodies (ER, PR, Her2) and/or their detection systems 
you will need to run more extensive test. As Liz says, 40 positive, 40 
negative. Again, a tissue array is great for that. 

Changing instruments these days is a big deal when a vendor ties you to their 
reagents. 

Tim Morken
Supervisor, Histology, IPOX
UCSF Medical Center
San Francisco, CA, USA

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz Chlipala
Sent: Tuesday, February 08, 2011 3:20 PM
To: Weems, Joyce; Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

Nope that's the recommendation for everything, in the paper they state
additional development is required for prognostic markers. Once you have
validated an antibody it only requires 3 tissues when you get a new lot
number:  1 strong positive, 1 weak to moderate positive and 1 negative.
>From how I read the article its 25 tissues and then 3 tissues, it does
not talk about slides so it is possible to put multiple tissues on one
slide.  Again these are just recommendations; I do not think that there
is a set standard yet.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949 
fax (303) 682-9060
www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: Weems, Joyce [mailto:jwe...@sjha.org] 
Sent: Tuesday, February 08, 2011 4:13 PM
To: Liz Chlipala; Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

But that is for receptors, correct? Do you do that for everything? 
Thanks, j 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz
Chlipala
Sent: Tuesday, February 08, 2011 17:58
To: Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

Joe

If you are following the recommendations from the CAP paper on IHC
standardization then it would be 25 tissues (10 strong positive, 10 weak
to moderate positive and 5 negative). 

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC
PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303)
682-9060 www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joe
Nocito
Sent: Tuesday, February 08, 2011 3:44 PM
To: Histonet
Subject: [Histonet] IHC validation

Greetings Histoland,
I need some help. We are about to switch IHC machines from the
Richard-Allen Axiom to the Ventana Benchmark Ultra. How many slides, per
antibody, do you run for the validation study? We have over 100 primary
antibodies. Normally, when we work up a new antibody, we  start with a
titer. Once that is established, we run 10 cases to check for
specificity. Hopefully we can obtain cases that are really positive,
some weakly positive and some flat out negative. Once that is completed,
we run 10 different tissue types to check for any unexpected
cross-reactivity. 
The ultra holds 30 slides and we are receiving two machines. If we
run 10 slides/antibody, that's going to take a while, not to mention the
number of detection kits that will be used. Do you think 5
slides/antibody is sufficient? I emailed CAP last week for their take
and they never returned my email (I told my medical director to hold
their check for the year and see how fast they respond to that). Ah oh,
don't go down that road Joe, it's unhealthy. What are your thoughts?
Thanks

Joe (JTT)


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not

RE: [Histonet] IHC validation

[Liz Chlipala]

Nope that's the recommendation for everything, in the paper they state
additional development is required for prognostic markers. Once you have
validated an antibody it only requires 3 tissues when you get a new lot
number:  1 strong positive, 1 weak to moderate positive and 1 negative.
>From how I read the article its 25 tissues and then 3 tissues, it does
not talk about slides so it is possible to put multiple tissues on one
slide.  Again these are just recommendations; I do not think that there
is a set standard yet.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949 
fax (303) 682-9060
www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: Weems, Joyce [mailto:jwe...@sjha.org] 
Sent: Tuesday, February 08, 2011 4:13 PM
To: Liz Chlipala; Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

But that is for receptors, correct? Do you do that for everything? 
Thanks, j 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz
Chlipala
Sent: Tuesday, February 08, 2011 17:58
To: Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

Joe

If you are following the recommendations from the CAP paper on IHC
standardization then it would be 25 tissues (10 strong positive, 10 weak
to moderate positive and 5 negative). 

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC
PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303)
682-9060 www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joe
Nocito
Sent: Tuesday, February 08, 2011 3:44 PM
To: Histonet
Subject: [Histonet] IHC validation

Greetings Histoland,
I need some help. We are about to switch IHC machines from the
Richard-Allen Axiom to the Ventana Benchmark Ultra. How many slides, per
antibody, do you run for the validation study? We have over 100 primary
antibodies. Normally, when we work up a new antibody, we  start with a
titer. Once that is established, we run 10 cases to check for
specificity. Hopefully we can obtain cases that are really positive,
some weakly positive and some flat out negative. Once that is completed,
we run 10 different tissue types to check for any unexpected
cross-reactivity. 
The ultra holds 30 slides and we are receiving two machines. If we
run 10 slides/antibody, that's going to take a while, not to mention the
number of detection kits that will be used. Do you think 5
slides/antibody is sufficient? I emailed CAP last week for their take
and they never returned my email (I told my medical director to hold
their check for the year and see how fast they respond to that). Ah oh,
don't go down that road Joe, it's unhealthy. What are your thoughts?
Thanks

Joe (JTT)


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Confidentiality Notice:
This e-mail, including any attachments is the 
property of Catholic Health East and is intended 
for the sole use of the intended recipient(s).  
It may contain information that is privileged and 
confidential.  Any unauthorized review, use,
disclosure, or distribution is prohibited. If you are 
not the intended recipient, please delete this message, and 
reply to the sender regarding the error in a separate email.


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RE: [Histonet] IHC validation

[Weems, Joyce]

But that is for receptors, correct? Do you do that for everything? 
Thanks, j 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz Chlipala
Sent: Tuesday, February 08, 2011 17:58
To: Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

Joe

If you are following the recommendations from the CAP paper on IHC 
standardization then it would be 25 tissues (10 strong positive, 10 weak to 
moderate positive and 5 negative). 

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 
18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 
www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joe Nocito
Sent: Tuesday, February 08, 2011 3:44 PM
To: Histonet
Subject: [Histonet] IHC validation

Greetings Histoland,
I need some help. We are about to switch IHC machines from the Richard-Allen 
Axiom to the Ventana Benchmark Ultra. How many slides, per antibody, do you run 
for the validation study? We have over 100 primary antibodies. Normally, when 
we work up a new antibody, we  start with a titer. Once that is established, we 
run 10 cases to check for specificity. Hopefully we can obtain cases that are 
really positive, some weakly positive and some flat out negative. Once that is 
completed, we run 10 different tissue types to check for any unexpected 
cross-reactivity. 
The ultra holds 30 slides and we are receiving two machines. If we run 10 
slides/antibody, that's going to take a while, not to mention the number of 
detection kits that will be used. Do you think 5 slides/antibody is sufficient? 
I emailed CAP last week for their take and they never returned my email (I told 
my medical director to hold their check for the year and see how fast they 
respond to that). Ah oh, don't go down that road Joe, it's unhealthy. What are 
your thoughts?
Thanks

Joe (JTT)


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Histonet mailing list
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Histonet@lists.utsouthwestern.edu
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Confidentiality Notice:
This e-mail, including any attachments is the 
property of Catholic Health East and is intended 
for the sole use of the intended recipient(s).  
It may contain information that is privileged and 
confidential.  Any unauthorized review, use,
disclosure, or distribution is prohibited. If you are 
not the intended recipient, please delete this message, and 
reply to the sender regarding the error in a separate email.


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RE: [Histonet] IHC validation

[Liz Chlipala]

Joe

If you are following the recommendations from the CAP paper on IHC
standardization then it would be 25 tissues (10 strong positive, 10 weak
to moderate positive and 5 negative). 

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949 
fax (303) 682-9060
www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joe
Nocito
Sent: Tuesday, February 08, 2011 3:44 PM
To: Histonet
Subject: [Histonet] IHC validation

Greetings Histoland,
I need some help. We are about to switch IHC machines from the
Richard-Allen Axiom to the Ventana Benchmark Ultra. How many slides, per
antibody, do you run for the validation study? We have over 100 primary
antibodies. Normally, when we work up a new antibody, we  start with a
titer. Once that is established, we run 10 cases to check for
specificity. Hopefully we can obtain cases that are really positive,
some weakly positive and some flat out negative. Once that is completed,
we run 10 different tissue types to check for any unexpected
cross-reactivity. 
The ultra holds 30 slides and we are receiving two machines. If we
run 10 slides/antibody, that's going to take a while, not to mention the
number of detection kits that will be used. Do you think 5
slides/antibody is sufficient? I emailed CAP last week for their take
and they never returned my email (I told my medical director to hold
their check for the year and see how fast they respond to that). Ah oh,
don't go down that road Joe, it's unhealthy. What are your thoughts?
Thanks

Joe (JTT)


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RE: [Histonet] IHC Validation (again)

[Liz Chlipala]

Laurie

I'm not aware of a particular question, but I would believe you would
have to perform some validation steps for each antibody.  I would
approach it the same way you approach validating new lots of antisera.
The CAP paper on standardization of IHC recommends 25 different samples
when you initially validate an antibody - 10 samples that have high
levels of target antigen, 10 intermediate to low levels and 5 negative.
To revalidate new lots they recommend only 3 tissue samples - 1 high, 1
med to low, and 1 negative.  Granted this only applies to routine
markers.  For prognostic markers such as ER/PR and Her2 then additional
samples need to be tested - there are new guidelines for ER/ER out and
the new recommendations for validation for ER/ER are briefly reviewed in
the CAP Today April issue.  Guidelines for validation of ER/PR will be
published in the June issue of Archives of Pathology and Laboratory
Medicine.

Liz 

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949 
fax (303) 682-9060
www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Laurie
Colbert
Sent: Wednesday, May 19, 2010 2:01 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC Validation (again)

Can anyone tell me if there is a specific question on the CAP checklist
that addresses revalidation of antibodies when starting up a new IHC
stainer?

 

Laurie Colbert

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RE: [Histonet] IHC Validation on new instrument

[Liz Chlipala]

We perform our entire validation process as a new piece of equipment.
Our validation protocols are quite extensive, up to about 85 pages long
on each piece of major equipment, at least that's what it was for our
new prisma stainer and glass coverslipper.  We perform an
installation/operational qualification protocol or an IOQ.  If we move
the instrument we also do the same thing, we already have the protocol
written which takes most of the time we just execute it again.  We are a
GLP lab so we work off a Validation Master Plan that basically tells us
how we are going to validate each piece of equipment in the lab.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949 
fax (303) 682-9060
www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Laurie
Colbert
Sent: Wednesday, May 19, 2010 9:00 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC Validation on new instrument

What do others do when validating a new model of a piece of equipment -
same manufacturer, same basic staining process, but an updated version
of the equipment?

 

I've been told the protocols should be the same and that we only need to
run three controls with three different but similar protocols to
determine what looks best.  Do you all think that is thorough enough, or
would you run actual patient cases and compare old and new equipment?  I
don't see where the CAP checklist refers to new equipment - just new
antibodies and new antibody lots.

 

Laurie Colbert

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