IMC pipeline based on the 26-marker panel. (A) The workflow of IMC. Continuous tissue sections of 18 patients with DLBCL were analyzed by IMC, H&E stain, and IHC. Protein expression of functional markers analyzed by IHC was to confirm the results from IMC. Patient prognosis based on differential expression of functional markers was retrospectively studied later in this study. (B) Representative IMC images showed the distribution of various markers. Scale bar, 200 μm. (C) Heat map of cellular phenotypes. A heat map represents major cell types identified within the TME. (D) UMAP plots were based on the single-cell data extracted from IMC images; 8 clusters from samples collected from 18 patients with DLBCL were defined according to their marker expression. (E) Representative images showed the distribution of cell type markers in UMAP plots. UMAP, uniform manifold approximation and projection.