Bildgebung der humanen immunologischen Synapse
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1Instituto de Investigaciones Biomédicas Alberto Sols,CSIC-Universidad Autónoma de Madrid2Departamento de Bioquímica, Instituto de Investigaciones Biomédicas Alberto Sols CSIC-UAM, Facultad de Medicina,Universidad Autónoma de Madrid3Unidad de Audiovisuales, Facultad de Medicina,Universidad Autónoma de Madrid
Summary
Abstract
Introduction
Protocol
Representative Results
Discussion
Disclosures
The authors have nothing to disclose.
Acknowledgements
Materials
| Camera Nikon DS-QI1MC | Nikon | MQA11550 | Cooled Camera Head |
| CMAC | ThermoFisher Scientific | C2110 | Cell tracker blue |
| JURKAT cells | ATCC | ATCC TIB-152 | Effector T lymphocytes |
| μ-Slide 8 well ibiTreat, μ-Slide 8 well Glass-Bottom | IBIDI | Cat.No: 80826, 80827 | Cell culture and cell imaging supports |
| Microscope NIKON Eclipse Ti-E | Nikon | NIKON Eclipse Ti-E | Wide-field fluorescence, fully-motorized microscope equipped with Perfect Focus System (PFS) option |
| Microscope Stage Incubator with 3-channel manual gas mixer and gas bubbler/ humidity module | OKOLAB | H201-NIKON-TI-S-ER | Cell culture atmosphere |
| Raji Cells | ATCC | ATCC CCL-86 | APC |
| RPMI medium GIBCO | ThermoFisher Scientific | 21875034 | Culture medium |
| Streptococcus Enterotoxin E (SEE) | Toxin Technology, Inc | EP404 | Bacterial Toxin |
| Software Huygens Essential | SVI | Huygens Essential | Image Deconvolution software |
| Software ImageJ | NIH | Image J | Image software |
| Software Nikon NIS-AR | Nikon | NIS-Elements AR | Image capture and analysis software |
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Tags
Immunological SynapseCell-to-cell ConjugationAntigen-presenting CellT-helper LymphocyteImmune Synapse FormationTime-lapse AcquisitionWide-field Fluorescence MicroscopySignal-to-noise RatioImmunofluorescence StainingFibronectinRaji CellsCell Tracker BlueMicroscopy TechniquesFluorescence BleachingPolarized Secretion
Cite This Article
Bello-Gamboa, A., Izquierdo, J. M., Velasco, M., Moreno, S., Garrido, A., Meyers, L., Palomino, J. C., Calvo, V., Izquierdo, M. Imaging the Human Immunological Synapse. J. Vis. Exp. (154), e60312, doi:10.3791/60312 (2019).