Texas Childrens Hospital and Baylor College of Medicine 1 article published in JoVE Immunology and Infection Visualization of the Immunological Synapse by Dual Color Time-gated Stimulated Emission Depletion (STED) Nanoscopy Emily M. Mace1, Jordan S. Orange1 1Center for Human Immunobiology, Texas Children's Hospital and Baylor College of Medicine 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.