PDCD1 and carcinoma: Despite this variable cell expression during our staining of mIF carcinoma tissues batches, overall we obtained good reproducibility and convenient staining with successful detection using PD-L1, CD3, CD8, CD57, and PD-1 markers compared with chromogenic IHC staining, and our group successfully multiplexed these biomarkers in two different panels by following our protocol, demonstrating the practical scalability of this method.