File:Live-cell-imaging-techniques-to-study-T-cell-trafficking-across-the-blood-brain-barrier-in-vitro-2045-8118-10-7-S1.ogv

English: Movie 1. Shear resistant arrest, polarization, crawling and diapedesis of CD4+ T cells on and across TNF-α stimulated wt pMBMECs under flow (low magnification). CD4+ T cells were perfused over TNF-α stimulated pMBMECs under low shear (0.1 dyn/cm2) (upper timer). After 4 min, flow was increased to physiological shear stress (1.5 dyne/cm2) (lower timer). The number of arrested CD4+ T cells constantly increased during the accumulation phase. Physiological shear washed away unbound T cells. Only few arrested CD4+ T cells detached from the endothelial surface whereas the majority of CD4+ T cells either continuously crawled or crawled and diapedesed through the endothelium. Phase-contrast bright T cells crawl on the apical surface of the endothelium, whereas phase-contrast dark T cells crawl beneath the endothelium. Direction of flow is from left to right. Objective 10x (Objective EC “Plan-Neofluar” 10x/0,3 Ph1 M27), phase-contrast illumination at 12 images per min, recording time 19 min. Movie at 12 images per sec, field of view 653 μm x 869 μm.