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. This evidence concerns the gene CD274 and carcinoma.