Design and application of an 8-plex multiplex immunofluorescence panel for deep phenotypic profiling of the tumor microenvironment using InSituPlex technology

An 8-plex InSituPlex® panel was engineered to profile T-cell, macrophage, and tumor phenotypes across NSCLC, CRC, and DCIS tissues, revealing key immune signatures and PD-L1 expression patterns at single-cell resolution. The workflow integrates high-throughput staining with whole-slide spatial analysis to uncover deeper relationships between exhausted, regulatory, and cytotoxic T-cell populations within the tumor microenvironment.