Re: [Histonet] Cell block processing
There's been excellent discussion on fixation, IHC and cells blocks, with Joe Walker's email summing up the situation nicely. For those interested in side by side comparison of IHC staining of cytology cell blocks using different fixatives go to http://cpqa.ca/main/wp-content/uploads/2014/06/2014-Thomson.pdf Regards John Garratt www.ciqc.ca ‐‐‐ Original Message ‐‐‐ On Monday, October 28, 2019 7:38 AM, Joe W. Walker, Jr. via Histonet wrote: > Hi Terri, > > At one time we did the same thing but have changed our approach in light of > the FDA's and CAP's view point on ASRs. The potential problem is that IHCs > are all validated/tested by the manufacturer on FFPE tissue. By introducing > methanol/ethanol as the first step in fixation, you potentially have altered > the initial fixation steps. I've attended several meetings on this topic and > have been advised to stop performing IHC on methanol/ethanol fixed specimens > unless we validated that this fixation step doesn't alter the expression of > the target antigen in the tissue. Formalin fixation after an alcohol fixation > doesn't change/reverse any alterations to the antigen in the tissue. > > We utilize an IBF tissue fixative but have also validated this fixative with > our antibody panels that we offer. The IBF does contain a small amount of > alcohol and the fixative is slightly different than 10% buffered formalin. > > I agree that CytoLyt is excellent at lysing red blood cells but would just > caution you on using the specimen for IHC without a disclaimer within your > report or validating your IHCs on these specimens to ensure they work as > expected. Keep in mind that most control tissue is FFPE and using it to > compare if the IHC worked in a first fixed alcohol specimen is not an apples > to apples comparison. > > Cheers, > > Joe W. Walker, Jr. MS, SCT(ASCP) > Anatomical Pathology Manager > joewal...@rrmc.org, www.rrmc.org > > -Original Message- > From: Terri Braud via Histonet histonet@lists.utsouthwestern.edu > Sent: Monday, October 28, 2019 10:14 AM > To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Cell block processing > > [External Email] This email originated from outside of the organization. > Think before you click: Don’t click on links, open attachments or respond to > requests for sensitive information if the email looks suspicious or you don’t > recognize the sender. > > We collect our FNAs in CytoLyt. We also use it to wash all our non-gyn > fluids, but then we fix the cell block "pellet" in formalin. > We have had no problems with immunos, and are able to lyse the RBCs to > provide a nice, clear specimen. > Hope this helps. > > 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 > > Histonet mailing list > Histonet@lists.utsouthwestern.edu > https://nam02.safelinks.protection.outlook.com/?url=https%3A%2F%2Furldefense.com%2Fv3%2F__http%3A%2F%2Flists.utsouthwestern.edu%2Fmailman%2Flistinfo%2Fhistonet__%3B!5JlSGkcjda6Eqs5J!rHP2_dFONxwRmR87ddavejT_o6RIcXBah8jw7nMUO0tWdC5KpksUnk2bttyIMkU%24&data=02|01|jwwalker%40rrmc.org|eeac9b3afe444de27cd208d75bb1634b|0e55647d438e4a448437e959c3cf2240|0|0|637078689748526729&sdata=1%2BpvxH9TcvKxAo6jgjr7fobCAUUz35sHbsLfoBvHmGE%3D&reserved=0 > [https://www.rrmc.org/app/files/public/2633/2019_hyht_sig-_jan2019_final.jpg] > > 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] PLAG1
Looking at an article in Histopathology (January 2018) on PLAG1 IHC testing, the authors used clone "3B7" (Novus Biologicals in Littleton, CO) on the Bond Max IHC platform form. Richard Richard W. Cartun, MS, PhD Director, Histology & The Martin M. Berman, MD Immunopathology/Morphologic Proteomics Laboratory 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: Charles Riley via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Monday, October 28, 2019 10:20 AM To: Histo List Subject: [Histonet] PLAG1 EXTERNAL email is from outside HHC. DO NOT open attachments or click links from unknown senders. My pathologists want to bring this test in house. Can anyone suggest a company I can purchase this antibody for testing via IHC on the Leica Bond platform? -- Charles Riley BS HT, HTL(ASCP)CM Histopathology Coordinator/ Mohs ___ Histonet mailing list Histonet@lists.utsouthwestern.edu https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.utsouthwestern.edu_mailman_listinfo_histonet&d=DwICAg&c=e_HtEeZEQXP5NUOb33qoTj0AVvRFBS9_rhBTQcfkWoA&r=eCMS5E4UqfGKTlPknIMjMlCkk-KGKHflslRsFz3l5JE&m=4VoB34evfULROSYTZirfyVjGhHd9khhhTv1g4pUQmxY&s=jqmKMH0dL56TvREZ4Edi5Oza2RVUTXB-PoqLdT5Sc2k&e= Reminder: This e-mail and any attachments are subject to the current HHC email retention policies. Please save or store appropriately in accordance with policy. 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] Cell block processing
Hi Terri, At one time we did the same thing but have changed our approach in light of the FDA's and CAP's view point on ASRs. The potential problem is that IHCs are all validated/tested by the manufacturer on FFPE tissue. By introducing methanol/ethanol as the first step in fixation, you potentially have altered the initial fixation steps. I've attended several meetings on this topic and have been advised to stop performing IHC on methanol/ethanol fixed specimens unless we validated that this fixation step doesn't alter the expression of the target antigen in the tissue. Formalin fixation after an alcohol fixation doesn't change/reverse any alterations to the antigen in the tissue. We utilize an IBF tissue fixative but have also validated this fixative with our antibody panels that we offer. The IBF does contain a small amount of alcohol and the fixative is slightly different than 10% buffered formalin. I agree that CytoLyt is excellent at lysing red blood cells but would just caution you on using the specimen for IHC without a disclaimer within your report or validating your IHCs on these specimens to ensure they work as expected. Keep in mind that most control tissue is FFPE and using it to compare if the IHC worked in a first fixed alcohol specimen is not an apples to apples comparison. Cheers, Joe W. Walker, Jr. MS, SCT(ASCP) Anatomical Pathology Manager joewal...@rrmc.org, www.rrmc.org -Original Message- From: Terri Braud via Histonet Sent: Monday, October 28, 2019 10:14 AM To: 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] Cell block processing [External Email] This email originated from outside of the organization. Think before you click: Don’t click on links, open attachments or respond to requests for sensitive information if the email looks suspicious or you don’t recognize the sender. We collect our FNAs in CytoLyt. We also use it to wash all our non-gyn fluids, but then we fix the cell block "pellet" in formalin. We have had no problems with immunos, and are able to lyse the RBCs to provide a nice, clear specimen. Hope this helps. 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu https://nam02.safelinks.protection.outlook.com/?url=https%3A%2F%2Furldefense.com%2Fv3%2F__http%3A%2F%2Flists.utsouthwestern.edu%2Fmailman%2Flistinfo%2Fhistonet__%3B!5JlSGkcjda6Eqs5J!rHP2_dFONxwRmR87ddavejT_o6RIcXBah8jw7nMUO0tWdC5KpksUnk2bttyIMkU%24&data=02%7C01%7Cjwwalker%40rrmc.org%7Ceeac9b3afe444de27cd208d75bb1634b%7C0e55647d438e4a448437e959c3cf2240%7C0%7C0%7C637078689748526729&sdata=1%2BpvxH9TcvKxAo6jgjr7fobCAUUz35sHbsLfoBvHmGE%3D&reserved=0 [https://www.rrmc.org/app/files/public/2633/2019_hyht_sig-_jan2019_final.jpg] ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Cell block processing
We collect our FNAs in CytoLyt. We also use it to wash all our non-gyn fluids, but then we fix the cell block "pellet" in formalin. We have had no problems with immunos, and are able to lyse the RBCs to provide a nice, clear specimen. Hope this helps. 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] PLAG1
My pathologists want to bring this test in house. Can anyone suggest a company I can purchase this antibody for testing via IHC on the Leica Bond platform? -- Charles Riley BS HT, HTL(ASCP)CM Histopathology Coordinator/ Mohs ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] BRAF KRAS NRAS by PCR
Does anyone do these tests using PCR on FFPE blocks? If so what platform do you use/recommend? -- Charles Riley BS HT, HTL(ASCP)CM Histopathology Coordinator/ Mohs ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
