RE: [Histonet] how to bleach melanoma cases without bad effect on IHC
We have actually performed a bleach step before staining. The only affect I have seen ( and we have only done a few antibodies) is that the tissue gets pretty chewed up. Especially if there is a pretreatment step prior to the IHC staining. Loralee McMahon, HTL (ASCP) Immunohistochemistry Supervisor Strong Memorial Hospital Department of Surgical Pathology (585) 275-7210 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cynthia Pyse [cp...@x-celllab.com] Sent: Tuesday, March 22, 2011 10:01 AM To: 'Shahram Sabeti'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] how to bleach melanoma cases without bad effect on IHC Sabeti I use the AEC chromogen (Dako catalog #K346989) for my melanoma cases. I use a aqueous mounting media, placed on the tissue only, dry it completely on a hot plate. When dry I coverslip it with permanent mounting media(don't place it in xylene), dry coverslip. Works great, and the color doesn't fade. Cindy Cindy Pyse, CLT, HT (ASCP) Laboratory/Histology Supervisor X-Cell Laboratories 716-250-9235 e-mail cp...@x-celllab.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Shahram Sabeti Sent: Tuesday, March 22, 2011 4:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] how to bleach melanoma cases without bad effect on IHC hello dear fellows, i have an IHC project on paraffin embedded melanoma cases; some of them being heavily pigmented. i have seen many protocols based on H2O2 to bleach the pigmented ones, but i am not sure about any possible effect on antigen retrieval.please guide me . yours, sabeti ___ 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] how to bleach melanoma cases without bad effect on IHC
Sabeti I use the AEC chromogen (Dako catalog #K346989) for my melanoma cases. I use a aqueous mounting media, placed on the tissue only, dry it completely on a hot plate. When dry I coverslip it with permanent mounting media(don't place it in xylene), dry coverslip. Works great, and the color doesn't fade. Cindy Cindy Pyse, CLT, HT (ASCP) Laboratory/Histology Supervisor X-Cell Laboratories 716-250-9235 e-mail cp...@x-celllab.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Shahram Sabeti Sent: Tuesday, March 22, 2011 4:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] how to bleach melanoma cases without bad effect on IHC hello dear fellows, i have an IHC project on paraffin embedded melanoma cases; some of them being heavily pigmented. i have seen many protocols based on H2O2 to bleach the pigmented ones, but i am not sure about any possible effect on antigen retrieval.please guide me . yours, sabeti ___ 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] how to bleach melanoma cases without bad effect on IHC
Sabeti, If you have the option of using a detection system with a chromogen other than DAB, i.e. Alkaline Phosphatase Red, that is by far the easiest way to get around the pigment issue. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Shahram Sabeti Sent: Tuesday, March 22, 2011 3:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] how to bleach melanoma cases without bad effect on IHC hello dear fellows, i have an IHC project on paraffin embedded melanoma cases; some of them being heavily pigmented. i have seen many protocols based on H2O2 to bleach the pigmented ones, but i am not sure about any possible effect on antigen retrieval.please guide me . yours, sabeti ___ 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
