Co-Detection of RNA and Protein to Explore Tumor-Immune Interactions Utilizing RNAscope With Imaging Mass Cytometry

RNAscope™ HiPlex assay can be combined with Imaging Mass Cytometry™ (IMC™) to enable same-slide co-detection of RNA and protein targets on FFPE tissues. IMC utilizes CyTOF^® technology for simultaneous assessment of 40-plus protein markers at subcellular resolution without spectral overlap or background autofluorescence, thus providing unprecedented insight into tissue organization and function. The RNAscope HiPlex v2 assay can detect up to 12 targets on FFPE tissues. By using metal-conjugated labeled probes instead of the Fluoros ( Fluoros T1-T4, Fluoro T5-T7, Fluori T8-T12), RNA targets can be detected using the Hyperion Imaging Mass Cytometry system.

This technical note provides a workflow and procedure to achieve RNA and protein co-detection data in IMC experiments. This workflow utilizes the highly sensitive and specific RNAscope technology for RNA detection with the multiplexing capability of IMC to visualize key RNA and protein markers in the same FFPE samples.