Fig. 1: Single-cell spatial resolution of NSCLC using imaging mass cytometry.

A Schematic depicting the acquisition and analysis of IMC multiplexed images from 102 LUAD and 102 LUSC cores. Created in BioRender. Quail, D. (2025) https://BioRender.com/k14v431. B Distribution of cell types in the tumour microenvironment and corresponding clinical variables (predominant histological pattern, stage, sex, age, body mass index (BMI), smoking status, pack years, and progression status) in LUAD (n = 102) and LUSC (n = 102). Defined cell types are in order of prevalence for each histological subtype, sorted by CD4+ T cell frequency for LUAD and neutrophil frequency for LUSC. C Representative images of multiplex antibody staining and paired segmented images for LUAD and LUSC. Scale bars = 100 μm. Legend for cell types can be seen in B. D Prevalence of total immune, myeloid, and lymphoid cells as % total cells across LUAD (n = 102) and LUSC (n = 102) cores. Two-tailed unpaired multiple comparison adjusted Student’s t tests; mean ± s.e.m. E Lymphoid infiltrate as % of total cells in male (56 cores; 28 patients) and female (46 cores; 23 patients) LUAD patients. Two-tailed unpaired Student’s t test; mean ± s.e.m. F Lymphoid infiltrate as % of total cells in younger (under 75 years, 68 cores; 34 pts) and older (over 75 years, 34 cores; 17 pts) LUAD patients. Two-tailed unpaired Student’s t test; mean ± s.e.m. G Comparison of cell frequencies between LUAD (n = 102) and LUSC (n = 102) cores for stromal cells, neutrophils and macrophages, other myeloid and lymphoid cells. Two-tailed unpaired multiple comparison adjusted Student’s t tests; mean ± s.e.m. H Proportion of each immune cell as % of total immune cells in LUAD (n = 102) cores from most to least prevalent; mean ± s.e.m. I Proportion of each immune cell as % total immune cells in LUSC (n = 102) cores from most to least prevalent; mean ± s.e.m. J Proportion of Ki67+ cancer cells across LUAD (n = 102) and LUSC (n = 102) cores. Two-tailed unpaired Student’s t test; mean ± s.e.m. P value = 0.00000271. K Proportion of pERK+ CD4+ and CD8+ T cells across LUAD (n = 102 and n = 102) and LUSC (n = 99 and n = 101) cores. Two-tailed unpaired Student’s t test; mean ± s.e.m. pERK+ CD4 + : P value = 0.0000248. pERK+ CD8 + : P value = 0.0000177. L Proportion of CD40 + B cells across LUAD (n = 101) and LUSC (n = 93) cores. Two-tailed unpaired Student’s t test; mean ± s.e.m. M Proportion of Arg1+ macrophages across LUAD (n = 102) and LUSC (n = 102) cores. Two-tailed unpaired Student’s t test; mean ± s.e.m. P value = 0.00000733. N Proportion of alpha-cleaved H3+ neutrophils across LUAD (n = 102) and LUSC (n = 102) cores. Two-tailed unpaired Student’s t test; mean ± s.e.m. P value = 0.000000000318. LUAD lung adenocarcinoma, LUSC lung squamous cell carcinoma, Cl Monocyte classical monocyte, Non-Cl Monocyte non-classical monocyte, Int Monocyte intermediate monocyte, NK cell Natural Killer cell, Treg regulatory T cell, CyTOF cytometry by time of flight, Arg1 Arginase 1.