Live cell barcoding enables the combined investigation of tumor- and immune cell phenotypes and interplays. (A) Single-cell suspensions were prepared from a lung tumor tissue biopsy and stimulated in vitro for different periods of time with the indicated reagents. Tumor as well as immune cells from individual conditions were then live cell barcoded, combined and stained for immune-relevant markers (Table S1). (B) Different immune as well as tumor cell populations were identified via the indicated gating scheme and analyzed for their expression of multiple surface markers. APCs = antigen presenting cells. (C) Combined data was subjected to SPADE clustering (100 clusters) and represented as a minimal spanning tree (MST). Selected clusters are overlaid with a color-scheme representing the normalized fold-change in unstimulated vs. the indicated stimulus.