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197), whereas excellent agreement was achieved at cut-offs of 50% (κ=0.908) and 5% (κ=0.823). DIA on mIF showed that PD-L1 commonly colocalised with CD68 macrophages and CD8 cytotoxic cells were closer to PD-L1 /CK tumour cells (TCs) than to PD-L1 /CK TCs in spatial distribution. A combination of mIF and DIA is useful for the quantification of PD-L1 expression and IC populations in NSCLC. Further validation of TPS at a cut-off of 1% and assay harmonisation is essential for translating this method in a diagnostic setting. A combination of