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1Instituto de Investigaciones Biomédicas Alberto Sols,CSIC-Universidad Autónoma de Madrid, 2Departamento de Bioquímica, Instituto de Investigaciones Biomédicas Alberto Sols CSIC-UAM, Facultad de Medicina,Universidad Autónoma de Madrid, 3Unidad de Audiovisuales, Facultad de Medicina,Universidad Autónoma de Madrid
Chapters
- 00:05Title
- 01:26Adhesion of Raji Cells to the Chamber Slides and 7-amino-4-chloromethylcoumarin (CMAC, Cell Tracker Blue) Labeling
- 03:31Pulse of CMAC labeled Raji Cells with Staphylococcal Enterotoxin E and Preparation of Jurkat Cells
- 04:51Co-seeding of Raji and Jurkat Cells and Time Lapse Imaging of Emerging Synaptic Conjugates
- 05:51End Point Formation of Synaptic Conjugates and Fixation
- 06:38Results: Imaging the Human Immunological Synapse
- 08:28Conclusion
This protocol images both the immunological synapse formation and the subsequent polarized secretory traffic towards the immunological synapse. Cellular conjugates were formed between a superantigen-pulsed Raji cell (acting as an antigen-presenting cell) and a Jurkat clone (acting as an effector helper T lymphocyte).
