Focusing solely on malignant cancer cells and the genes that they express is modestly just a glimpse of the larger picture. The tumor microenvironment (TME) plays an important role in both tumor progression and the potential treatment response; identifying the key players involved in this environment could further reveal the immune system’s potential to regulate cancer. To achieve this, we used RNAscope® Assay to visualize immune checkpoint markers in the TME of non-small cell lung carcinoma biopsies from patients. Detection of co-expressed markers revealed a diverse pattern, highlighting intratumor heterogeneity within each tumor. Information derived from this study may reveal potential insight into combination therapies targeted against different checkpoint pathways.