Joanne What Dako platform are you using, link or 48? On the link I pretreat with PK, the IS series antibody is used with an incubation time of 10 minutes.. On the 48 I pretreat with High pH TRS, the IR antibody is used with an incubation time of 20 minutes. Both systems give me clean staining.(I'm in the process of phasing out the link.) I would try either decreasing the incubation time of the antibody if you stay with the HIER pretreatment or try the enzyme pretreatment with your current protocol. Good luck. Cindy
Cindy Pyse, CLT, HT (ASCP) Laboratory Manager X-Cell Laboratories 716-250-9235 Ext. 232 e-mail [email protected] -----Original Message----- From: [email protected] [mailto:[email protected]] On Behalf Of Joanne Clark Sent: Tuesday, February 28, 2012 11:28 AM To: [email protected] Subject: [Histonet] Cytokeratin AE1/AE3 Hi Histonetters, I am having some issues with my Cytokeratin AE1/AE3. We use the antibody from DAKO and do HIER on it before staining. We run AE1/AE3 as a protocol on sentinel lymph nodes from breast cases that were negative on frozen section to rule out micro metastases and what my pathologists sometimes sees is staining that looks like it's running in-between the cells. It is not an indication of metastases but my pathologists want me to get rid of it. Have any of you seen anything like this? I would especially appreciate the opinion of Samuri Pathologist on this if it isn't too much trouble. Thanks all! Joanne Clark, HT Histology Supervisor Pathology Consultants of New Mexico Roswell _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
