File:A-Novel-Cervical-Spinal-Cord-Window-Preparation-Allows-for-Two-Photon-Imaging-of-T-Cell-video 5.ogv
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[edit]DescriptionA-Novel-Cervical-Spinal-Cord-Window-Preparation-Allows-for-Two-Photon-Imaging-of-T-Cell-video 5.ogv |
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. |
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Source | Video S5 from Haghayegh Jahromi N, Tardent H, Enzmann G, Deutsch U, Kawakami N, Bittner S, Vestweber D, Zipp F, Stein J, Engelhardt B (2017). "A Novel Cervical Spinal Cord Window Preparation Allows for Two-Photon Imaging of T-Cell Interactions with the Cervical Spinal Cord Microvasculature during Experimental Autoimmune Encephalomyelitis". Frontiers in Immunology. DOI:10.3389/fimmu.2017.00406. PMID 28443093. PMC: 5387098. | ||
Author | Haghayegh Jahromi N, Tardent H, Enzmann G, Deutsch U, Kawakami N, Bittner S, Vestweber D, Zipp F, Stein J, Engelhardt B | ||
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This file is licensed under the Creative Commons Attribution 4.0 International license.
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current | 15:19, 20 May 2017 | 8.7 s, 1,046 × 1,046 (2 MB) | Open Access Media Importer Bot (talk | contribs) | Automatically uploaded media file from Open Access source. Please report problems or suggestions here. |
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Author | Haghayegh Jahromi N, Tardent H, Enzmann G, Deutsch U, Kawakami N, Bittner S, Vestweber D, Zipp F, Stein J, Engelhardt B |
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Usage terms | http://creativecommons.org/licenses/by/4.0/ |
Image title | 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. |
Software used | Xiph.Org libtheora 1.1 20090822 (Thusnelda) |
Date and time of digitizing | 2017-04-11 |