You can use DAB but the issue of endogenous peroxidase will rear its ugly head.
And I suppose IF being around since 1955, when Mellors first applied the technique to renal tissue, it has a long history of diagnostic application that is hard to replace. There are other enzymes that could be used that, not being present in human tissue, would not require endogenous enzyme blocking for example glucose oxidase. The advantage of this enzyme is that there is no endogenous glucose oxidase activity in mammalian tissues. Weening, J. J., & Jennette, J. C. (2012). Historical milestones in renal pathology. Virchows Archiv, 461(1), 3-11. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children’s Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA From: Allan Wang [mailto:alla...@gmail.com] Sent: Tuesday, 18 April 2017 5:51 PM To: Tony Henwood (SCHN) Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] help!! Tim and Tony, Why couldn't DAB be used on frozen sections in your example? Allan On Mon, Apr 17, 2017 at 9:03 PM, Tony Henwood (SCHN) via Histonet <histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>> wrote: Hi Bianca, Well for most Pathology departments, Immunofluorescence (IF) is used for Renal and Skin biopsies; looking for human Immunoglobulin (Igs) deposition on basement membranes. The advantage here is not so much the fluorescence, but that we use unfixed frozen sections. The buffer rinse before antibody application, removes un-bound serum immunoglobulins, leaving any pathological bound Igs for the IF antibody to bind to. This gives a clean result. If one would do IF on formalin-fixed paraffin sections of renal or skin biopsies, you would find heavy background due to the fixative cross-linking serum Igs to tissue and cells (which would usually be removed by the buffer rinse if unfixed frozen sections were used - see above). IF, apart from being a historic method, also does not suffer from endogenous peroxidase that would need to be blocked if peroxidase was used in place of fluorescence. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: Blanca Lopez via Histonet [mailto:histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>] Sent: Thursday, 13 April 2017 11:10 PM To: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu> Subject: [Histonet] help!! Hello! I just need a help with a simple question...Is anyone can explain me what is the purpose between performing immunohistochemistry and Immunofluorescence? Thanks :) Blanca Lopez Histotech (ASCP) UTSW Tissue Resource K1.210 Simmons Comprehensive Cancer Center UT Southwestern Medical Center Telephone: 214-648-7598<tel:214-648-7598> Email: blanca.lo...@utsouthwestern.edu<mailto:blanca.lo...@utsouthwestern.edu> ________________________________ UT Southwestern Medical Center The future of medicine, today. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu<mailto:Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message are those of the individual sender, and are not necessarily the views of NSW Health or any of its entities. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu<mailto:Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message are those of the individual sender, and are not necessarily the views of NSW Health or any of its entities. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
