Re: [Histonet] Antibody Validation CLIA
Cap states, "should test a minimum of 10 positive and 10 negative tissues". Does not state "cases" but "tissues", so we used a block for positive and negative from the same case. Thanks, Tim -Original Message- From: histonet-requ...@lists.utsouthwestern.edu [mailto:histonet-requ...@lists.utsouthwestern.edu] Sent: Friday, March 16, 2018 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 172, Issue 13 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-requ...@lists.utsouthwestern.edu You can reach the person managing the list at histonet-ow...@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Leica ST5050 immunostainer (Matthew Fleming) 2. Antibody Validation CLIA (Paula) -- Message: 1 Date: Fri, 16 Mar 2018 08:00:00 -0500 From: Matthew Fleming To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Leica ST5050 immunostainer Message-ID: Content-Type: text/plain; charset="UTF-8" Hi, I have a small dermatopathology lab and would like to introduce limited immunohistochemistry. Initially I would probably just do MITF stains with red chromogen on a few cases per week. We could do this by hand but I've noticed a number of ST5050 immunostainers for sale at very low prices. I know they're quite old but we have a number of machines old or older that continue to serve us well. But so far I haven't been able to find a manual and am concerned about finding consumables. Leica apparently stopped supporting this model some time ago. Is anyone out there still using it? Anyone have a manual? Anyone like to comment as to whether it might be usable for a limited IH operation? Thank you, Matthew Fleming, MD Fleming Dermatopathology Milwaukee, WI -- Message: 2 Date: Fri, 16 Mar 2018 06:54:30 -0700 From: "Paula" To: Subject: [Histonet] Antibody Validation CLIA Message-ID: <001801d3bd2e$4f4b0c20$ede12460$@biopath.org> Content-Type: text/plain; charset="us-ascii" 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 -- Subject: Digest Footer ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- End of Histonet Digest, Vol 172, Issue 13 * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Antibody Validation CLIA
Keep in mind like Lacy said its cases and not slides so you could place multiple cases on one slide or create a simple array using disposable biopsy punches. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com<mailto:l...@premierlab.com> www.premierlab.com<http://www.premierlab.com/> Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 From: Normington Lacy via Histonet Sent: Friday, March 16, 2018 11:17 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Antibody Validation CLIA CAP suggests running 10 negative and 10 positive cases for non-prognostic markers. CAP required running 20 negative and 20 positive cases for prognostic markers. In the event the case volume is less than the suggested 10 and 10 cases for non-prognostic markers, the reason for that decision should be stated in the validation. Ultimately, the decision is up to your director. Lacy Normington, HTL(ASCP)CM Manager, Surgical Pathology Lab Services UW Health 600 Highland Avenue Madison, WI 53792-2472 -Original Message- From: Paula via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Friday, March 16, 2018 8:55 AM To: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu> Subject: [Histonet] Antibody Validation CLIA WARNING: This email appears to have originated outside of the UW Health email system. DO NOT CLICK on links or attachments unless you recognize the sender and know the content is safe. 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<mailto:Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu<mailto: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] Antibody Validation CLIA
Hi Paula, Let me first say I am Canadian and my lab is not governed by CLIA or CAP. But for what it is worth, here are my thoughts... When thinking about validating antibodies for IHC you must first consider whether the antibody in question is Class-I (prognostic eg Breast markers, CD117, etc) or Class-II. Class-I require a more robust validation protocol then Class-II antibodies. Next you need to consider why the validation is needed. Is it: 1. a new Ab in your lab? 2. a new vendor for an antibody? 3. a new lot number of an existing antibody in your menu? 4. a change in protocol of an existing antibody? #'s 1 and 2 require a larger validation whereas #'s 3 and 4 should only require a minimal check (unless of course there is a drastic change in the protocol in #4). As for absolute numbers of slides for each, unless CLIA provides this information, you are left to work with your Director to find that balance between cost (ie more slides = increased cost) with confidence (how many do you need to see in order to feel confident that the stain is performing as expected). I hope this helps. Sincerely, 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
Re: [Histonet] Antibody Validation CLIA
CAP suggests running 10 negative and 10 positive cases for non-prognostic markers. CAP required running 20 negative and 20 positive cases for prognostic markers. In the event the case volume is less than the suggested 10 and 10 cases for non-prognostic markers, the reason for that decision should be stated in the validation. Ultimately, the decision is up to your director. Lacy Normington, HTL(ASCP)CM Manager, Surgical Pathology Lab Services UW Health 600 Highland Avenue Madison, WI 53792-2472 -Original Message- From: Paula via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Friday, March 16, 2018 8:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Antibody Validation CLIA WARNING: This email appears to have originated outside of the UW Health email system. DO NOT CLICK on links or attachments unless you recognize the sender and know the content is safe. 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] ANtibody validation records
Here is the document our developmental specialist uses when bring on new antibodies. Hope this helps. Lacy Normington Lacy Normington, HTL(ASCP)CM Manager, Surgical Pathology Lab Services 600 Highland Avenue Madison, WI 53792-2472 Phone: 608-890-9373 -Original Message- From: Charles Riley via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Thursday, February 16, 2017 7:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] ANtibody validation records Would anyone be willing to share how the document their new antibody validations when adding a new antibody test to the panel of tests your lab performs. I currently have a basic packet but feel it could be better and wanted to see how everyone else performs/records their testing. The people who were in my position before me failed to keep track of all the new antibodies and now I am trying to redo them all before our upcoming CAP inspection. Any help would great be appreciated -- Charles Riley HT(ASCP)CM Histopathology Coordinator/ Mohs ___ 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] ANTIBODY VALIDATION PROCEDURE
Follow the CAP guidelines for antibody validation; that's what we do. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Wilson A Sent: Wednesday, May 16, 2012 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] ANTIBODY VALIDATION PROCEDURE Hi, How are you guys doing? I hope you are doing great. Please I will appreciate it, if you guys have a procedure on the ANTIBODY VALIDATION and would like to share it with me. Thanks Wilson ___ 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] Antibody Validation
Funny, I'd think the vendors would want you to do tons of your own validation, as it uses up antibody faster, ergo they get to sell you more antibody! :) In a feisty mood today. Claire From: histonet-boun...@lists.utsouthwestern.edu on behalf of Rene J Buesa Sent: Tue 6/15/2010 9:49 AM To: histo...@pathology.swmed.edu; teri.hall...@midmichigan.org Subject: Re: [Histonet] Antibody Validation Teri: You are right about the validations you propose although I am not surprised that your vendor does not think it is necessary. They are in the business of selling and you are in the business of assuring the high quality of your work to obtaining the most accurate work for patients' sake. There is where the difference resides. Ignore your vendor and keep validating your protocols. René J. --- On Tue, 6/15/10, teri.hall...@midmichigan.org wrote: From: teri.hall...@midmichigan.org Subject: [Histonet] Antibody Validation To: histo...@pathology.swmed.edu Date: Tuesday, June 15, 2010, 7:55 AM I am being questioned by our vendor as to why we need to validate our automated immunostainer and image analysis instrument. They would like documentation pertaining to the requirement of validation and the number of specimens utilized for validation. I am requesting that each antibody be validated on the instrument against a previously validated instrument. Additionally, I am requesting that each new lot of antibody be validated upon receipt against previously ran specimens. This would also apply to the image analysis antibodies. (Her2 has been validated by FISH.) The vendor has apparently polled users in the area and this is not a standard protocol, therefore the request for documentation. I think it is pretty clearly stated by CAP in the Quality Management In Anatomic Pathology. Any other suggestions? Teresa Hallada BS, MT/CT (ASCP) Pathology Lead MidMichigan Health - Gratiot teri.hall...@midmichigan.org 989.463.1101 ext 3423 ___ Please note that this email message and any attachments may contain privileged and confidential information that is protected against use or disclosure under federal and state law. The information is intended only for the personal and confidential use of the intended recipient. If the reader of this message is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this information in error and that any review, dissemination, distribution, copying or action taken in reliance on the contents of this communication is strictly prohibited. If you have received this email in error, please advise by immediate reply. ___ 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] Antibody Validation - long response
Well said Elizabeth Jesus A Ellin HT/PA ASCP Department of Pathology/Histology Yuma Regional Medical Center 2400 South Ave A Yuma, AZ 85364 - 7170 ( Office: (928) 336-1743 (Fax: (928) 336-7319 *Email: jel...@yumaregional.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz Chlipala Sent: Tuesday, June 15, 2010 9:01 AM To: Rene J Buesa; histo...@pathology.swmed.edu; teri.hall...@midmichigan.org Subject: RE: [Histonet] Antibody Validation - long response Bottom line it's not the vendors responsibility to validate their equipment or antibodies in your lab. Some vendors may help you do this, but ultimately the lab needs to validate the equipment and IHC in their lab. The vendors normally calibrate the equipment prior to shipment and once they set the instrument up in your lab, they should be able to provide you with the documentation that states that they calibrated the instrument. Your instruments need to be calibrated prior to being validated. As far as your scanner goes some vendors can provide validation, but it's at a cost and that cost is not cheap depending upon what you actually want validated. If you are using the scanner and associated algorithms for analysis then you need to validate that separately. There are several steps required to validate a scanner - 1. you validate the scanner 2. if you are using a database to store your images then that also may need to be validated and 3. if you are using algorithms that provide you with data then those algorithms need to be validated. For example prior to running a validation protocol on a tissue processor its needs to be calibrated for temperature. All of your major equipment needs to be on a calibration schedule. We calibrate all of our instruments once a year and validation is completed only once unless we change the instrument location or how we use the instrument. Pipettors are calibrated every 6 months. All instruments are validated it may just be a one pager for the basic lab equipment but instruments like the tissue processor, slide staining, IHC stainer and scanner require written protocols some of these are 80 pages in length and go into great detail. The same goes for your antibodies. Antibodies are validated initially with 25 tissue samples (10 strongly positive tissues, 10 moderate to weakly positive tissues and 5 tissues that have no reactivity) This type of validation is required for routine antibodies, prognostic markers such as Her-2, ER and PR require additional tissue samples. New lots require 3 tissue samples one strongly positive on moderate to weakly positive and one negative. If you change the antibody source or detection system or retrieval it needs to be validated again - This information comes from the paper Standarization of Immunohistochemistry from CAP its available on line - I have a copy if you need it. There are also new guidelines for ER/PR and a new article on validation of ER/PR in the June issue of Archives of pathology from CAP. 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 Rene J Buesa Sent: Tuesday, June 15, 2010 8:50 AM To: histo...@pathology.swmed.edu; teri.hall...@midmichigan.org Subject: Re: [Histonet] Antibody Validation Teri: You are right about the validations you propose although I am not surprised that your vendor does not think it is necessary. They are in the business of selling and you are in the business of assuring the high quality of your work to obtaining the most accurate work for patients' sake. There is where the difference resides. Ignore your vendor and keep validating your protocols. René J. --- On Tue, 6/15/10, teri.hall...@midmichigan.org wrote: From: teri.hall...@midmichigan.org Subject: [Histonet] Antibody Validation To: histo...@pathology.swmed.edu Date: Tuesday, June 15, 2010, 7:55 AM I am being questioned by our vendor as to why we need to validate our automated immunostainer and image analysis instrument. They would like documentation pertaining to the requirement of validation and the number of specimens utilized for validation. I am requesting that each antibody be validated on the instrument against a previously validated instrument. Additionally, I am requesting that each new lot of antibody be validated upon receipt against previously ran specimens. This would also apply to the image analysis antibodies. (Her2 has been validated by FISH.) The vendor has apparently polled users in the area and this is not a standard protocol, theref
RE: [Histonet] Antibody Validation - long response
Bottom line it's not the vendors responsibility to validate their equipment or antibodies in your lab. Some vendors may help you do this, but ultimately the lab needs to validate the equipment and IHC in their lab. The vendors normally calibrate the equipment prior to shipment and once they set the instrument up in your lab, they should be able to provide you with the documentation that states that they calibrated the instrument. Your instruments need to be calibrated prior to being validated. As far as your scanner goes some vendors can provide validation, but it's at a cost and that cost is not cheap depending upon what you actually want validated. If you are using the scanner and associated algorithms for analysis then you need to validate that separately. There are several steps required to validate a scanner - 1. you validate the scanner 2. if you are using a database to store your images then that also may need to be validated and 3. if you are using algorithms that provide you with data then those algorithms need to be validated. For example prior to running a validation protocol on a tissue processor its needs to be calibrated for temperature. All of your major equipment needs to be on a calibration schedule. We calibrate all of our instruments once a year and validation is completed only once unless we change the instrument location or how we use the instrument. Pipettors are calibrated every 6 months. All instruments are validated it may just be a one pager for the basic lab equipment but instruments like the tissue processor, slide staining, IHC stainer and scanner require written protocols some of these are 80 pages in length and go into great detail. The same goes for your antibodies. Antibodies are validated initially with 25 tissue samples (10 strongly positive tissues, 10 moderate to weakly positive tissues and 5 tissues that have no reactivity) This type of validation is required for routine antibodies, prognostic markers such as Her-2, ER and PR require additional tissue samples. New lots require 3 tissue samples one strongly positive on moderate to weakly positive and one negative. If you change the antibody source or detection system or retrieval it needs to be validated again - This information comes from the paper Standarization of Immunohistochemistry from CAP its available on line - I have a copy if you need it. There are also new guidelines for ER/PR and a new article on validation of ER/PR in the June issue of Archives of pathology from CAP. 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 Rene J Buesa Sent: Tuesday, June 15, 2010 8:50 AM To: histo...@pathology.swmed.edu; teri.hall...@midmichigan.org Subject: Re: [Histonet] Antibody Validation Teri: You are right about the validations you propose although I am not surprised that your vendor does not think it is necessary. They are in the business of selling and you are in the business of assuring the high quality of your work to obtaining the most accurate work for patients' sake. There is where the difference resides. Ignore your vendor and keep validating your protocols. René J. --- On Tue, 6/15/10, teri.hall...@midmichigan.org wrote: From: teri.hall...@midmichigan.org Subject: [Histonet] Antibody Validation To: histo...@pathology.swmed.edu Date: Tuesday, June 15, 2010, 7:55 AM I am being questioned by our vendor as to why we need to validate our automated immunostainer and image analysis instrument. They would like documentation pertaining to the requirement of validation and the number of specimens utilized for validation. I am requesting that each antibody be validated on the instrument against a previously validated instrument. Additionally, I am requesting that each new lot of antibody be validated upon receipt against previously ran specimens. This would also apply to the image analysis antibodies. (Her2 has been validated by FISH.) The vendor has apparently polled users in the area and this is not a standard protocol, therefore the request for documentation. I think it is pretty clearly stated by CAP in the Quality Management In Anatomic Pathology. Any other suggestions? Teresa Hallada BS, MT/CT (ASCP) Pathology Lead MidMichigan Health - Gratiot teri.hall...@midmichigan.org 989.463.1101 ext 3423 ___ Please note that this email message and any attachments may contain privileged and confidential information that is protected against use or disclosure under federal and state law. The information is intended only for the personal and confidential use of the intended re
RE: [Histonet] Antibody Validation
There are other issue that you have to take into consideration with both instruments, With digital image analysis you also have to look at the questions that CAP just instituted for this. This includes the machine being run by someone that can do high complexity testing. This means that you have to have a tech do this machine, not a TA or Lab aid. There are also issue with consistent calibration of the Image analysis machines, continued education of the techs, and validating not only anitbodies but continued validation of the image algorythms.. Do not listen to vendors, rely on your instinct and validate. Jesus A Ellin HT/PA ASCP Department of Pathology/Histology Yuma Regional Medical Center 2400 South Ave A Yuma, AZ 85364 - 7170 ( Office: (928) 336-1743 (Fax: (928) 336-7319 *Email: jel...@yumaregional.org __ This message is confidential, intended only for the named recipient(s) and may contain information that is privileged or exempt from disclosure under applicable law. If you are not the intended recipient(s), you are notified that the dissemination, distribution, or copying of this message is strictly prohibited. If you receive this message in error, or are not the named recipient(s), please notify the sender at either the e-mail, fax, address, or telephone number listed above and delete this e-mail from your computer. Thank You. __ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Antibody Validation
I agree with Rene. All lot to lot and new antibodies need to be checked for consistency. This is part of your validation process. This even holds true for pre-dilutes which are already tested by the manufacturer for optimum results. Beatrice Sullivan, HT(A.S.C.P.) HTL , AAS, CLSP(N.C.A.) AP Supervisor Shore Memorial Hospital 609-653-3590 Rene J Buesa To Sent by: histo...@pathology.swmed.edu, histonet-bounces@ teri.hall...@midmichigan.org lists.utsouthwest cc ern.edu Subject Re: [Histonet] Antibody Validation 06/15/2010 10:49 AM Teri: You are right about the validations you propose although I am not surprised that your vendor does not think it is necessary. They are in the business of selling and you are in the business of assuring the high quality of your work to obtaining the most accurate work for patients' sake. There is where the difference resides. Ignore your vendor and keep validating your protocols. René J. --- On Tue, 6/15/10, teri.hall...@midmichigan.org wrote: From: teri.hall...@midmichigan.org Subject: [Histonet] Antibody Validation To: histo...@pathology.swmed.edu Date: Tuesday, June 15, 2010, 7:55 AM I am being questioned by our vendor as to why we need to validate our automated immunostainer and image analysis instrument. They would like documentation pertaining to the requirement of validation and the number of specimens utilized for validation. I am requesting that each antibody be validated on the instrument against a previously validated instrument. Additionally, I am requesting that each new lot of antibody be validated upon receipt against previously ran specimens. This would also apply to the image analysis antibodies. (Her2 has been validated by FISH.) The vendor has apparently polled users in the area and this is not a standard protocol, therefore the request for documentation. I think it is pretty clearly stated by CAP in the Quality Management In Anatomic Pathology. Any other suggestions? Teresa Hallada BS, MT/CT (ASCP) Pathology Lead MidMichigan Health - Gratiot teri.hall...@midmichigan.org 989.463.1101 ext 3423 ___ Please note that this email message and any attachments may contain privileged and confidential information that is protected against use or disclosure under federal and state law. The information is intended only for the personal and confidential use of the intended recipient. If the reader of this message is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this information in error and that any review, dissemination, distribution, copying or action taken in reliance on the contents of this communication is strictly prohibited. If you have received this email in error, please advise by immediate reply. ___ 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] Antibody Validation
Teri: You are right about the validations you propose although I am not surprised that your vendor does not think it is necessary. They are in the business of selling and you are in the business of assuring the high quality of your work to obtaining the most accurate work for patients' sake. There is where the difference resides. Ignore your vendor and keep validating your protocols. René J. --- On Tue, 6/15/10, teri.hall...@midmichigan.org wrote: From: teri.hall...@midmichigan.org Subject: [Histonet] Antibody Validation To: histo...@pathology.swmed.edu Date: Tuesday, June 15, 2010, 7:55 AM I am being questioned by our vendor as to why we need to validate our automated immunostainer and image analysis instrument. They would like documentation pertaining to the requirement of validation and the number of specimens utilized for validation. I am requesting that each antibody be validated on the instrument against a previously validated instrument. Additionally, I am requesting that each new lot of antibody be validated upon receipt against previously ran specimens. This would also apply to the image analysis antibodies. (Her2 has been validated by FISH.) The vendor has apparently polled users in the area and this is not a standard protocol, therefore the request for documentation. I think it is pretty clearly stated by CAP in the Quality Management In Anatomic Pathology. Any other suggestions? Teresa Hallada BS, MT/CT (ASCP) Pathology Lead MidMichigan Health - Gratiot teri.hall...@midmichigan.org 989.463.1101 ext 3423 ___ Please note that this email message and any attachments may contain privileged and confidential information that is protected against use or disclosure under federal and state law. The information is intended only for the personal and confidential use of the intended recipient. If the reader of this message is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this information in error and that any review, dissemination, distribution, copying or action taken in reliance on the contents of this communication is strictly prohibited. If you have received this email in error, please advise by immediate reply. ___ 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
Fw: Re: [Histonet] antibody validation Thanks, but need Non-"U" also
Hi Cynthia, Thank you, this is exactly what I was looking for. You are a rare jewel. Especially for the size of your facility. I am impressed! I wish I would have received more feed-back from small labs, but that's OK. I have been doing some pro-active sleuthing of local labs. After numerous conversations with my fellow histologists that do IHC, I am finding out what I have long suspected and have experienced while doing technical support and consulting. The routine histology laboratory is multitasking all of their regular histology duties with the addition of IHC testing. Pathologists do not want to lose the IHC revenue to a reference laboratory, so they bring on-board simple IHC panels, and send out their esoteric or prognostic markers to a well known reference laboratory. Most histologists have been asked to bring on board IHC, without the proper educational background or tools. The average histologist has not had the opportunity to take continuing education classes, or even go on the histonet. This may be due to lack of continuing education funds, or lack of interest; It is amazing that pathologists can go to their continuing education meetings, but don't support their histologists. Unfortunately, these histologists do not understand the importance of standardization or validation steps. When something goes bump in the night, they are unaware how to troubleshoot the problem. This is unfortunate for the histologist, the pathologist and most of all, the patient. One more reason for pathologists to support continuing education. Hope to see you all at NSH! Akemi Allison-Tacha BS, HT(ASCP)HTL Histology Manager APMG Laboratories 105A Cooper Court, Los Gatos, CA 95032 Contact: 800.848.2764 V/M: 408.884.2718 Fax: 408.884.2758 Cell: 408.335.9994 (W) E-Mail: aallison-ta...@apmglab.com (P) E-Mail: akemiat3...@yahoo.com --- On Mon, 9/14/09, Cynthia Robinson wrote: From: Cynthia Robinson Subject: Re: [Histonet] antibody validation Thanks, but need Non-"U" also To: "Akemi Allison-Tacha" Date: Monday, September 14, 2009, 11:08 AM Hi, I work at 250 bed facility. For the initial protocol workup we use the manufacturer recommended protocol and run an IHC TMA control block with 25 different tissues and tumors. Once the pathologist has looked and responded I search for 5-10 cases with expected positive results and then a selection of 5-10 normal or negative tissues and run the protocol. Hope this is what you were wanting. Cindi Cindi Robinson HT(ASCP) Mercy Medical Center Dunes Medical Laboratories 350 W Anchor Dr Dakota Dunes SD 57049 phone-712-279-2768 robin...@mercyhealth.com >>> Akemi Allison-Tacha 9/14/2009 12:53 PM >>> Hi All, I would like to thank those of you who have responded to the e-mail below regarding antibody validation. The information I received was pretty much what I expected from university IHC labs. I would like to know what smaller IHC labs are doing for validation and re-validation when changes are made to their protocols. Thank you again for any and all responses, Akemi Akemi Allison-Tacha BS, HT(ASCP)HTL Histology Manager APMG Laboratories 105A Cooper Court, Los Gatos, CA 95032 Contact: 800.848.2764 V/M: 408.884.2718 Fax: 408.884.2758 Cell: 408.335.9994 (W) E-Mail: aallison-ta...@apmglab.com (P) E-Mail: akemiat3...@yahoo.com --- On Fri, 9/11/09, Akemi Allison-Tacha wrote: From: Akemi Allison-Tacha Subject: [Histonet] antibody validation To: "histonet" Date: Friday, September 11, 2009, 8:20 AM Good Morning and Happy Friday out there in Histo-Land! I would like your assistance in an issue that I have just become aware of regarding antibody validation. I have been requested to bring on-board approximately (7) new antibodies in the immediate future, and several antibodies later. Although, many of you who know me realize I have worked in IHC R&D for a well known IHC company for many years, it has been a while since I have overseen a clinical IHC lab. I would like your assistance and input on how you are validating new antibodies. Not counting Her2-neu, it has been my understanding that it is recommended to test samples with varying antigen expression from 10-15 cases. Furthermore, you should check for Sensitivity: expression range of Positive cases, low to high (10-15 cases). Specificity: (+) vs expected (-) cases/tissues (10-15 cases) and Precision (Reproducibility): It has been my practice that any changes that are made to existing protocols, such as a new detection kit or antigen retrieval methodology, also requires re-validation. I am curious how you approach validation. Also, do any of you just use a single known positive control for that antibody, and run it, and if it is positive, feel that it is satisfactory, and put it on-line for testing. Thank you in advance for your input, and have a great weekend! Akemi Allison-Tacha BS, HT(
RE: [Histonet] antibody validation
Our procedure is: Read the data sheet for the antibody (recommended dilution, any pre-treatment) Read references that use the same antibody/clone (what should stain, what internal controls, their staining conditions) Obtain a known, or suspected, positive control and titre the antibody using recommended pre-treatment. Start with the titre recommended by the data sheet or from the literature. Using the optimised conditions stain a variety of tissues or lesions that should be positive as well as those lesions that should be negative (especially those that enter into the differential diagnosis) Record the results in your lab notebook and YOU ARE Done but, Continue to monitor the antibody's performance - this is important. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Akemi Allison-Tacha Sent: Saturday, 12 September 2009 1:21 AM To: histonet Subject: [Histonet] antibody validation Good Morning and Happy Friday out there in Histo-Land! I would like your assistance in an issue that I have just become aware of regarding antibody validation. I have been requested to bring on-board approximately (7) new antibodies in the immediate future, and several antibodies later. Although, many of you who know me realize I have worked in IHC R&D for a well known IHC company for many years, it has been a while since I have overseen a clinical IHC lab. I would like your assistance and input on how you are validating new antibodies. Not counting Her2-neu, it has been my understanding that it is recommended to test samples with varying antigen expression from 10-15 cases. Furthermore, you should check for Sensitivity: expression range of Positive cases, low to high (10-15 cases). Specificity: (+) vs expected (-) cases/tissues (10-15 cases) and Precision (Reproducibility): It has been my practice that any changes that are made to existing protocols, such as a new detection kit or antigen retrieval methodology, also requires re-validation. I am curious how you approach validation. Also, do any of you just use a single known positive control for that antibody, and run it, and if it is positive, feel that it is satisfactory, and put it on-line for testing. Thank you in advance for your input, and have a great weekend! Akemi Allison-Tacha BS, HT(ASCP)HTL Histology Manager APMG Laboratories 105A Cooper Court, Los Gatos, CA 95032 Contact: 800.848.2764 V/M: 408.884.2718 Fax: 408.884.2758 Cell: 408.335.9994 (W) E-Mail: aallison-ta...@apmglab.com (P) E-Mail: akemiat3...@yahoo.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet * 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 are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] antibody validation
Akemi, We follow pretty much what you outline below. I believe following strict and comprehensive validation will save you headaches later on. You want to 1) convince yourself that the antibody works as advertised and as expected according to the literature and 2)Identify and solve any technical issues that may cause spurious results. You can't do that with a one-off test of one tissue sample. After a well-done validation you should have great confidence that it is being done right and works correctly. It is very useful to have the validation data in hand if anyone questions your test results. Suggested references: Recommendations for Improved Standardization of Immunohistochemistry, Goldstein, NS, et.al., and members of Ad-Hoc Committee on Immunohistochemical Standardization, Appl Immunohistochem Mol Morph, 2007 15(2): 124-133 A Practical Approach for Evaluating New Antibodies in the Clinical Immunohistochemistry Laboratory, Hsi, ED, Arc Pathol Lab Med. 2001; 125: 289-294 Quality Assurance For Immuncytochemistry: Approved Guideline, Clinical Laboratory Standards Institute (formerly NCCLS), Wayne PA, USA, publication MM4-A, Vol. 19, No. 26, 1999. www.clsi.org Book chapters on IHC Validation and QC: Quality Management in Immunohistochemistry, Brown RW, in Quality Management in Anatomic Pathology, Nakhleh RE, and Fitzgibbons PL, Eds. College of American Pathologists, Northfield, IL, USA, 2005. www.cap.org. Theoretical and Practical Aspects of Test Performance, in Immunohistology: A Diagnostic Tool for the Surgical Pathologist. 3rd. Ed., Volume 19 in Major Problems in Pathology, Taylor CR and Cote RJ, Eds., W.B Saunders, Philadelphia, 2005 Immunohistochemistry Quality Control, Hladik, CL and White, CL, in Theory and Practice of Histological Techniques, 4th Ed., Bancroft JD and Gamble M, Eds., Churchill Livingstone, 2007 Techniques of Immunohistochemistry: Principles, Pitfalls and Standardization, Taylor CR, et.al., in Diagnostic Immunohistochemistry, Dabbs, DJ, Ed., 2nd Edition, Churchill Livingstone, Tim Morken Supervisor, Histology / IPOX UCSF Medical Center San Francisco, CA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Akemi Allison-Tacha Sent: Friday, September 11, 2009 8:21 AM To: histonet Subject: [Histonet] antibody validation Good Morning and Happy Friday out there in Histo-Land! I would like your assistance in an issue that I have just become aware of regarding antibody validation. I have been requested to bring on-board approximately (7) new antibodies in the immediate future, and several antibodies later. Although, many of you who know me realize I have worked in IHC R&D for a well known IHC company for many years, it has been a while since I have overseen a clinical IHC lab. I would like your assistance and input on how you are validating new antibodies. Not counting Her2-neu, it has been my understanding that it is recommended to test samples with varying antigen expression from 10-15 cases. Furthermore, you should check for Sensitivity: expression range of Positive cases, low to high (10-15 cases). Specificity: (+) vs expected (-) cases/tissues (10-15 cases) and Precision (Reproducibility): It has been my practice that any changes that are made to existing protocols, such as a new detection kit or antigen retrieval methodology, also requires re-validation. I am curious how you approach validation. Also, do any of you just use a single known positive control for that antibody, and run it, and if it is positive, feel that it is satisfactory, and put it on-line for testing. Thank you in advance for your input, and have a great weekend! Akemi Allison-Tacha BS, HT(ASCP)HTL Histology Manager APMG Laboratories 105A Cooper Court, Los Gatos, CA 95032 Contact: 800.848.2764 V/M: 408.884.2718 Fax: 408.884.2758 Cell: 408.335.9994 (W) E-Mail: aallison-ta...@apmglab.com (P) E-Mail: akemiat3...@yahoo.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
