File:A-Novel-Cervical-Spinal-Cord-Window-Preparation-Allows-for-Two-Photon-Imaging-of-T-Cell-video 7.ogv

English: Diapedesis of activated 2D2 GFP CD4+ T cells across inflamed cervical spinal cord post-capillary venules during experimental autoimmune encephalomyelitis (EAE). Laminectomy (described in step 15 of the procedures) was performed in a C57BL/6 mouse at the onset of EAE. In vitro activated CD4+ T cells from “2D2-GFP”-mice were systemically injected via a carotid catheter before 2P-IVM imaging. During the recording of 15 min, two CD4+ T cells undergoing diapedesis across cervical spinal cord post-capillary venules can be observed. A x–y–t time-lapse sequence of a 300 μm × 300 μm at a depth of 60–112 μm and 14 z-stacks with 4 µm spacing is shown. Blood vessels were labeled with Alexa Fluor 594 conjugated anti-endoglin antibody. GFP (green, CD4+ T cells) and Alexa Fluor 594 (red, blood vessels) were excited at 780 nm. Time is shown in minutes and seconds. Scale bar: 40 µm.