Carol, Are you doing the MSI by hand, or by autostainers (which platform?).
Here's my protocols that I have optimized; Protocol A for Dako Plus (or manually by hand); 1) AR; Biocare's Diva with Pascal pressure cooker for 3-5 min 2) Quench End. Peroxidase; Dako's End.H202 for 5 min 3) Incubate 1st Ab; PMS2 (Cell Marque; RTU) antibody for 60 min 4) Incubate 2nd Ab; Dako Envision +/HRP for 30 min 5) Develop color; Dako Envision DAB+ for 5 min 6) Counterstain; Mayer Hematoxylin for 2 min 7) Dehydrate & cover slip with permanent mounting Protocol B for Leica Bond; 1) Use Protocol F with Bond Refine detection kit 2) HIER; ER2 for 20 min My IHC staining are always strong and consistent (manually by hand, Dako Plus, or Leica Bond). Hope this help! Madeleine Huey BS, HTL (ASCP) QIHC Supervisor - Pathology (IPOX & Histology) El Camino Hospital madelein...@elcaminohospital.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet