Table 1.
Current Immunoscore procedure and reagents
| 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) |