Membrane Trafficking

Characterizing Membrane Traffic in the Early Secretory Pathway Using the RUSH Retention System.- Monitoring Intra-Golgi Transport with Acute Spatiotemporal Control of a Synthetic Cargo.- Imaging Secretory Granule Budding from the Trans-Golgi Network Using Retention Using Selective Hook (RUSH).- Spatial and Temporal Control of Protein Secretion with Light.- Generation of Endogenously Tagged Membrane Trafficking Regulators Using CRISPR Genome Editing.- Determine the Function of the Exocyst in Vesicle Tethering by Ectopic Targeting.- Imaging Insulin Granule Dynamics in Human Pancreatic beta-Cells Using Total Internal Reflection Fluorescence (TIRF) Microscopy.- Monitoring Synaptic Exocytosis Using a Whole-Cell Patch Clamp Technique.- Live-Cell Super-Resolution Imaging of Retrograde Axonal Trafficking Using Pulse-Chase Labeling in Cultured Hippocampal Neurons.- Electron Tomographic Methods for Studying Organelles of the Murine Chemical Synapse.- In Vitro Reconstitution Studies of SNAREs and Their Regulators Mediating GLUT4 Vesicle Fusion.- Imaging Single Vesicle Exocytosis with Total Internal Reflection Fluorescence Microscopy (TIRFM).- Localizing Proteins on Single Trafficking Organelles in 3D with Semisynthetic Gold Labeling and Platinum Replica Electron Microscopy.- Endocytosis Assays Using Cleavable Fluorescent Dyes.- Reconstituting and Purifying Assembly Intermediates of Clathrin Adaptors AP1 and AP2.- Monitoring Endosomal Cargo Retrieval to the Trans-Golgi Network by Microscopic and Biochemical Approaches.- Measuring Plasma Membrane Recycling Using Microscopic and Biochemical Approaches.- Light Scattering Techniques to Assess Self-Assembly and Hydrodynamics of Membrane Trafficking Proteins.- Measurement of Lysosome Positioning by Shell Analysis and Line Scan.- Optogenetic Control of Membrane Trafficking Using Light-Activated Reversible Inhibition by Assembly Trap of Intracellular Membranes (IM-LARIAT).- Generating Custom Pooled CRISPR Libraries for Genetic