Fig. 1: Digital spatial profiling (DSP) illuminates consistent profiles from distinct masks in lymphoid tissue microregions.

A Schematic of GeoMx® DSP WTA workflow (created with BioRender.com). B, C Immunofluorescence staining of DLBCL tissues (n = 87) and RLTs (n = 24). In Group 1, CD68 stained macrophages (yellow), CD3 stained T cells (cyan), CD20 stained B cells (magenta), and SYTO 13 stained nuclei (blue). In Group 2, CD68 stained macrophages (yellow), NGFR illuminated LZ (green) and SYTO 13 stains nuclei (blue). After ROI selection, each cell type was segmented based on the staining signal and their corresponding masks were generated. Representative images are shown. Scale bar: 100 μm. Source data are provided as a Source Data file. D, E Cumulative density functions showed that the signatures of macrophages (CD68, CD163, FCGR1A, and CSF1R), T cells (CD3D, CD3E, UBASH3A, CD2, and TRBC2), and B cells (MS4A1, CD79A, CD79B, CD19, and PAX5) were highly enriched in CD68+ regions, CD3+ regions, and CD20+ regions, respectively in RLTs and DLBCL tissues (Kolmogorov-Smirnov P < 0.05). Digital spatial profiling, DSP; whole transcriptome analysis, WTA; diffuse large B-cell lymphoma, DLBCL; reactive lymphoid tissues, RLTs; regions of interest, ROIs; areas of interest, AOIs; formalin-fixed paraffin-embedded, FFPE; light zone, LZ; dark zone, DZ; nerve growth factor receptor, NGFR.