Table 1 Current Immunoscore procedure and reagents
From: Cancer classification using the Immunoscore: a worldwide task force
Procedure | Current recommended steps |
|---|---|
Tumor selection | Block which is the most infiltrated by the immune cells and containing the core of the tumor (CT) and the invasive margin (IM) |
Sample preparation | 2 paraffin sections of 4-microns of the tumor block deposited in deionized water on Superfrost-plus slides |
Immuno-histochemistry (IHC) | 2 single stainings using IVD certified antibodies |
Antigen retrieval | CC1 tris-based buffer pH8 |
Primary antibody | CD3 (2GV6, Ventana) and CD8 (C8/144, Dako) |
Primary antibody diluant | K 004 (Clinisciences) for CD8 |
Secondary reagents | Ultraview TM DAB (Ventana) |
Counterstaining | Hematoxillin II (Ventana) |
Autostrainer | Benchmark XT (Ventana) |
Scanner | NanoZoomer 2.0-HT (Hammamatsu) |
Digital pathology | Architect XD software (Definiens) |
Immunoscore quantification | Immunoscore Plug-in (INSERM / AP-HP) |