[Histonet] Re: Histonet Digest, Vol 98, Issue 17

[Madeleine Huey]

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
madeleine_h <@t> elcaminohospital.org