Re: [Histonet] IHC Validation Question
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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC Validation Question
Morning Histo Friends. Got a quick question for you. I am getting ready to go over to a new start up lab that will be doing all IHC staining in house. I have set up a lab before and done validations but that was on equipment and special stains which I will be doing again and feel pretty good about it. Any tips and tricks for validating IHCs? We will be using the Benchmark Ultra (please no solicitation we are all set). We are hoping not to have to buy too many controls so I have been looking to try finding some in house I can use to help save the cost. If anyone has any good advice for this to I will be all ears. Thanks in advance. Phillip Kara Senior Research Associate UNT HSC ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC validation after a new tissue processer is installed
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**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC validation after a new tissue processer is installed
Hello all, After a very long time our histology department is getting a new tissue processor, and hopefully several more in the near future. Of course for IHC that means a revalidation/reverification of our IHC stains. Since I have not had to do this before I wanted to get some guidance on how to go about doing this. I feel like I have to completely revalidate the ER, PR and Her2 (20 positive, 20 negative) but wasn't sure about the other antibodies. We have a menu of around 130do I have to test every one of them or can we chose a representative sample? How many antibodies would you suggest? How many positive and negative cases of each should we run? It says it is left up to the medical director but with our CAP inspection next spring we want to make sure we will be fully prepared.What have others done? Thank you in advance for your help with this. Martha Ward, MT (ASCP) QIHC Wake Forest Baptist Medical Center ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC validation
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! ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[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! ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC validation
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 > > > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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 > > > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC validation
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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC validation
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 Protecting your privacy is important to us. The Baker Heart and Diabetes Institute will handle your information in accordance with the Privacy Act 1988 (Cth) and its Privacy Policy which is available at www.baker.edu.au or on request by contacting priv...@baker.edu.au or by calling 1800 838 498. The Privacy Policy also explains how you can access and correct your personal information, or make a complaint about a breach of the Australian Privacy Principles. bidipp2014.0.1a -- Message protected by MailGuard: e-mail anti-virus, anti-spam and content filtering.http://www.mailguard.com.au/mg ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC validation
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 > > > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC VALIDATION
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 > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC VALIDATION
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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC Validation
I am validating IHC's. What is the CAP requirements regarding negative slides. I've asked CAP but have not received a response. Are 10 negative patient tissues ran for each antibody using the negative protocol or are they ran using each antibody but also a known Negative control ran using the negative protocol? Thanks! Karla Arrington, HT (ASCP) HIT (AHIMA) Supervisor of Pathology ANMC Pathology kaarring...@anthc.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC Validation
To All: I am going to be setting an IHC lab. I would like any suggestions as far as a form for me to use for validating each antibody. I would like to use a sample form if possible if anyone would like to share. Thanks! Karla Arrington, HT(ASCP), HIT(AHIMA) Lead Histology Technician kaarring...@anthc.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC Validation:
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 > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC Validation:
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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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 > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC validation:
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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged 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 contact the sender by reply e-mail and destroy all copies of the original message, including any attachments. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC validation:
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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC Validation:
Does anyone know the preferred method for IHC validation? Is making a tissue microarray of different known cases the preferred method? Does anyone have experience validating like this? Please let me know if anyone has information/tips to doing so. Thank you. Sent from my iPhone ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC Validation
Can you forward any templates to me as well? Thanks! -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Leslie Graves Sent: Tuesday, July 15, 2014 8:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: IHC Validation Thank you to everyone who sent a template. This is very helpful. I do need clarfication on the 10 cases that are required for validation. Do you include the 10 cases on this same form, or do you have something separate for that? These look ideal for optimization. From what I have read online, 10 cases need to be run after optimization has been completed. When validating these 10 cases, accuracy, sensitivity, specificity and reproducibility need to be monitored. Are all of you using separate form to capture this data and approval from the Pathologists? Maybe I am going overboard, but I want to make sure that I have everything covered. We are bringing a new platform into our lab in 2 weeks, and I want to have everything ready to go. Leslie Graves Technical Specialist Cape Fear Valley Medical Center Fayetteville, NC 28341 910-615-6142 lg...@capefearvalley.com On Mon, Jul 14, 2014 at 1:55 PM, Leslie Graves wrote: > Does anyone have a sample IHC Validation worksheet that they used in > their lab for review. I know that the requirements have changed since > our last inspection. I want to make sure that I am including all of > the criteria, and would like to see how other labs are documenting this > process. > > Leslie Graves > Technical Specialist > Cape Fear Valley Medical Center > 1638 Owen Drive > Fayetteville, NC 28341 > 910-615-6142 > lg...@capefearvalley.com > ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC Validation
Does anyone have a sample IHC Validation worksheet that they used in their lab for review. I know that the requirements have changed since our last inspection. I want to make sure that I am including all of the criteria, and would like to see how other labs are documenting this process. Leslie Graves Technical Specialist Cape Fear Valley Medical Center 1638 Owen Drive Fayetteville, NC 28341 910-615-6142 lg...@capefearvalley.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC VALIDATION
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." > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > This e-mail message, including any attachments, is for the sole use of the > intended recipient(s) and may contain confidential and privileged > 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 > contact the sender by reply e-mail and destroy all copies of the original > message, including any attachments. > > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC VALIDATION
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." ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged 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 contact the sender by reply e-mail and destroy all copies of the original message, including any attachments. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[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." ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC Validation
If I have validated an antibody at a specific incubation time, and then later want to decrease the incubation time, do I have to re-validate?? Laurie Colbert, HT (ASCP) Histology Supervisor PATH MD 8158 Beverly Blvd. Los Angeles, CA 90048 (323) 648-3214 direct (424) 245-7284 main lab The information contained in this transmission may contain privileged and confidential information, including patient information protected by federal and state privacy laws. It is intended only for the use of the person(s) named above. If you are not the intended recipient, you are hereby notified that any review, dissemination, distribution, or duplication of this communication is strictly prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC validation
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<mailto: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>)" mailto:histonet@lists.utsouthwestern.edu>> Message-ID: <12ECD7346266D74691EC2BFC75285E451CC048EB@BFL323E10.pathmdlabs.local<mailto: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) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC validation
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) > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[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) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC Validation
-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 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 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 originator of the message. This footer also confirms that this email message has been scanned for the presence of computer viruses. Any views expressed in this message are those of the individual sender, except where the sender specifies and with authority, states them to be the views of Commonwealth Health. Scanning of this message and addition of this footer is performed by Websense Email Security software in conjunction with virus detection software. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC Validation
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 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 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 originator of the message. This footer also confirms that this email message has been scanned for the presence of computer viruses. Any views expressed in this message are those of the individual sender, except where the sender specifies and with authority, states them to be the views of Commonwealth Health. Scanning of this message and addition of this footer is performed by Websense Email Security software in conjunction with virus detection software. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information in this e-mail, and any attachments therein, is confidential and for use by the intended addressee only. If this message is received by you in error please do not disseminate or read further. Please reply to the sender that you have received the message in error, then delete the message. Although Catholic Health Services of Long Island attempts to sweep e-mail and attachments for viruses, it does not guarantee that either are virus-free and accepts no liability for any damage sustained as a result of viruses. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC Validation
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 originator of the message. This footer also confirms that this email message has been scanned for the presence of computer viruses. Any views expressed in this message are those of the individual sender, except where the sender specifies and with authority, states them to be the views of Commonwealth Health. Scanning of this message and addition of this footer is performed by Websense Email Security software in conjunction with virus detection software. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[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 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 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 originator of the message. This footer also confirms that this email message has been scanned for the presence of computer viruses. Any views expressed in this message are those of the individual sender, except where the sender specifies and with authority, states them to be the views of Commonwealth Health. Scanning of this message and addition of this footer is performed by Websense Email Security software in conjunction with virus detection software. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Ihc validation
I am having to validate antibodies and was wondering if you have to do a parallel study or just use known positive and negative tissue? ~Sabrina~ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC validation
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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC validation
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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC Validation
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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC Validation
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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC validation
Hematoxylin should not affect the immunoreactivity (since counterstaining occurs following the IHC reaction), so no, changing the hematoxylin would not require a revalidation. Just make sure you are not obscuring the immunostain with an overly-dark counterstain. The point is to provide contrast between the DAB (brown) or fast red chromagen and to offer the reader a sense of context of the surrounding morphology of the tissue being stained. Ashley Troutman BS, HT(ASCP) QIHC Immunohistochemistry Supervisor Vanderbilt University Histopathology 1301 Medical Center Drive TVC 4531 Nashville, TN 37232 Message: 10 Date: Tue, 19 Apr 2011 14:04:32 -0500 From: "Ring, Mary L" Subject: [Histonet] IHC validation To: "histonet@lists.utsouthwestern.edu" Message-ID: <2594d39b69223047b6e2f4adf1e62db9010ffb487...@hpemx3.healthpartners.int> Content-Type: text/plain; charset="iso-8859-1" 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC validation
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 This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC validation
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 This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC validation
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 This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC validation
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) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC validation
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) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC validation
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) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE:[Histonet] IHC validation
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) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC validation
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) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC validation
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
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
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) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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
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) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[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) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC Validation (again)
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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC Validation on new instrument
Hi Laurie, I have a Benchmark Ultra and a Benchmark XT from Ventana and they follow the basic steps, similar protocols and I needed to revalidate everything. They are sufficiently different (in my experience) that it warranted a complete revalidation. A part of the reasoning for this was some of the bulk reagents were different (a consideration that may lead you to not completely revalidate). For me, there were a few protocols that were quite different from one to the other. I would go to the trouble up front, that way there is no question later. Good luck! Ashley Troutman BS, HT(ASCP) QIHC Vanderbilt University Histopathology 1301 Medical Center Drive TVC 4532 Nashville, TN 37232 615-343-9134 Message: 4 Date: Wed, 19 May 2010 07:59:30 -0700 From: "Laurie Colbert" Subject: [Histonet] IHC Validation on new instrument To: Message-ID: <57be698966d5c54eae8612e8941d768308bca...@exchange3.huntingtonhospital.com> Content-Type: text/plain; charset="us-ascii" 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC Validation on microwaved tissue
Hello All, We have new microwave tissue processor, until this purchase all tissue has been placed on a standard VIP processor. My medical director would like for most of our 125 antibodies worked up on microwave processed tissue. I have been challenged with getting specimens to process on the instrument for IHC antibody validation purposes. Initially we will be processing GI biopsy specimens, but we will eventually progress to other biopsy tissues and so on. I have had success getting some colon cancer cases, some tonsil and other tissue. We will be mainly performing H. pylori, which is hard to get a large enough specimen to obtain a sample for this purpose. Anyway I was wondering how other labs are handling validation in this situation. Thank you for your time in this matter. Matt Brooks, BS, HT (ASCP) Histology Supervisor InCyte Pathology mbro...@incytepathology.com 509-892-2744 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC Validation on new instrument
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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC Validation Procedure
Can anyone share their IHC Validation procedure. I am preparing for a CAP inspection and want to make sure that the procedure I (presently do/may change to) is acceptable. Thank you very much! ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet