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Immunology and Infection

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Visualization of the Immunological Synapse by Dual Color Time-gated Stimulated Emission Depletion (STED) Nanoscopy

Article DOI: 10.3791/51100-v • 10:00 min • March 24th, 2014
March 24th, 2014 •
1, 1
1Center for Human Immunobiology, Texas Children's Hospital and Baylor College of Medicine

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Here we illustrate the protocol for imaging by two-color STED nanoscopy the cytotoxic immune synapse of NK cells recapitulated on glass. Using this method we obtain sub-100 nm resolution of synapse proteins and the cytoskeleton.

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Immunological Synapse Dual Color Time-gated Stimulated Emission Depletion (STED) Nanoscopy Natural Killer Cells Actin Reorganization Confocal Microscopy Fluorescence Microscopy Subdiffraction Limited Super-resolution Imaging F-actin Architecture Cytotoxic Synapse Activating Receptor Glass Substrate Proteins Of Interest Sample Preparation Image Acquisition Leica SP8 Software Huygens Software Post-processing Deconvolution
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