English: Intraluminal crawling of activated 2D2 CD4+ T cells in inflamed cervical spinal cord post-capillary venules during experimental autoimmune encephalomyelitis (EAE). In vitro activated 2D2 GFP CD4+ T cells and 2D2 CD4+ T cells labeled with fluorescent CellTracker CMAC were systemically injected via the carotid artery catheter into a surgically prepared EAE mouse at the onset of disease. A CMAC labeled T cell (blue cell) is seen to crawl against the direction of blood flow for the entire 20 min of recording. A GFP+ T cell is seen to crawl against the direction of blood flow until 3 min of the recording when it detached and re-entered circulation. Another GFP+ T cell (at time point of 13 min) and two additional CMAC labeled T cells (at time points of 8 min:40 s and 9 min:40 s) can be observed to transiently arrest on and crawl along the vascular wall and to rapidly re-enter blood circulation. A x–y–t time-lapse sequence of a 300 μm × 300 μm scan field at a depth of 52–115 μm and 16 z-stacks with 4.2 μm spacing is shown. Blood vessels were labeled by injection of Alexa Fluor 594 conjugated anti-endoglin antibody. GFP (green, CD4+ T cells), CMAC (blue, fluorescently labeled CD4+ T cells), and anti-endoglin (red, blood vessels) were excited at 780 nm. Time is shown in minutes and seconds. Scale bar: 30 μm.