Protocol for studying the immune microenvironment of human hepatocellular carcinoma by Cell DIVE multiplex immunofluorescence imaging.

Understanding tumor heterogeneity by examining how cells are organized in tissues is essential to better decipher the biological processes involved in cancer and to improve therapeutic management. Here, we present a protocol for a multiplex immunofluorescence (mIF) technique that enables the detection of an array of protein markers and allows immunophenotyping on a single section of human hepatocellular carcinoma. We describe the procedures for antibody labeling, mIF image acquisition using Cell DIVE, and preliminary analysis using QuPath software.