Spatial transcriptomics workflow on the GeoMx Digital Spatial Profiler. Within a delineated ROI, guided sampling of molecular barcodes from the unique fluorescence-segmented AOIs occurs within the instrument. These barcodes are bound to probes against nearly 18 676 RNAs by ultraviolet light-sensitive bonds. Sampling is nonoverlapping and sequential, ensuring spatial separation of probes from each AOI. A library is prepared from the extracted probes and sequenced, and raw data files are generated with information on number of RNA transcripts per gene for every AOI. Differential expression analysis and correlation studies follow, providing leads on disease subtypes and tissue neighborhood interactions. PanCK, pan cytokeratin.