Given that the in situ immune patterns of either PTs and metastases had prognostic and therapeutic implications, we used multiplex immunofluorescence and multispectral imaging to evaluate the in situ immune patterns of T cells (CD3+) and cytotoxic T cells (CD8+) in terms of their densities and locations to explore if there was a difference of in situ immune patterns between the PTs and the mLNs of NSCLC in this study. The gene discussed is CD8A; the disease is non-small cell lung carcinoma.