File:A-Novel-Cervical-Spinal-Cord-Window-Preparation-Allows-for-Two-Photon-Imaging-of-T-Cell-video 4.ogv

English: Intraluminal crawling of an activated 2D2 CD4+ T cell in inflamed cervical spinal cord post-capillary venules during experimental autoimmune encephalomyelitis (EAE). In vitro activated 2D2 CD4+ T-cells labeled with fluorescent CellTracker CMAC were systemically injected via the carotid artery catheter into a surgically prepared mouse at the onset of EAE. In the regions of interest, one transferred CD4+ T cell was observed to crawl along the direction of blood flow until 4 min and 20 s of recording. At this time, the T cell changed the direction of crawling against the blood flow until minute 12 of the recording. At this time point, the observed T cell again changed the direction of crawling along the direction of blood flow and continued to crawl to the end of 20 min of recording. A x–y–t time-lapse sequence of a 200 μm × 200 μm scan field at a depth of 79–100 μm and 8 z-stacks with 3 µm spacing is shown. Blood vessels were labeled by injection of Alexa Fluor 594 conjugated anti-endoglin antibody. CMAC (blue, CD4+ T cell) and anti-endoglin (red, blood vessels) were excited at 780 nm. Time is shown in minutes and seconds. Scale bar: 20 μm.