Immunohistochemical staining of paraffin-embedded Human Appendix triple stained for PD1 (Yellow) - CD8 (Teal) - CD103 (Purple)

Immunohistochemical staining of paraffin-embedded Human Appendix triple stained for PD1 (Yellow) - CD8 (Teal) - CD103 (Purple)

Multiplex (triple) immuno histochemical staining

Multiplex (triple) immuno histochemical staining is performed on a Discovery Ultra stainer (Ventana Medical Systems) using different colors of chromogenic substrates instead of fluorochromes. One of the advantages of using chromogenic substrates is that the morphology of the tissues stays visible.

Two Triple stainings (mentioned below) are set up to analyze the immune infiltration in Triple Negative Breast Cancer samples from the Young Boost Trial in collaboration with Hugo Horlings, Michiel de Maaker, Iris Nederlof, Marc van de Vijver, Rolf Harkens, Bram van den Broek, Siamak Hajizadeh and members of the CFMPB (Ingrid Hofland and Dennis Peters).

Triple Staining (chromogenic)                                      Autostainer
PDL1 - CD3 - CD68 Yellow - Teal - Purple                      Discovery Ultra
PD1 - CD8 - CD103 Yellow - Teal - Purple                      Discovery Ultra

Images are analyzed using different digital scoring methods. Either using our 3D-Histech P1000 scanner and FIJI/QuPath. Or using the Vectra 3.0 system to scan and extract each color separately, which are then analyzed using HALO.

 

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