Fig. 3

Single-cell tracking of 3D migration and invasion. Representative fluorescent images acquired with the Operetta CLS 48 h after the initiation of the 3D migration (A) and invasion (E) assays for OPBG-DIPG002 single-cell-derived clones 1C5 (Venus) and 2B4 (m-Orange2) either in mono or in co-culture (overlay of m-Orange2 and Venus) Scale Bar = 200 μm. For cell tracking experiments, images were acquired every 30 min with the Operetta CLS over 48 h. Single-cell tracking was performed using the Harmony software as represented with the lines and arrows overlayed on the fluorescent images and the mean cell displacement (B–F), speed (C–G) and accumulated distance (D–H) were determined as reported on the Graph bar. Data are mean ± SD, n = 3. (****) p < 0.0001; (***) p < 0.001; (**) p < 0.01; (*) p < 0.05