Membrane trafficking, i.e. endocytic/exocytic, endo-lysosomal and autophagic, pathways are well studied, this field has grown rapidly over the last decade. Recently a “new era of membrane trafficking diseases” as emerged and de-regulation of vesicle trafficking pathways was shown to be related to syndromes such as: Parkinson’s, Alzheimer’s, cancer, and metabolic syndromes like type2-diabetes.

Intracellular vesicles (e.g. endosomes and lysosomes) with sizes of a few hundred nanometres cannot be resolved by conventional confocal microscopy. Super-resolution microscopy techniques like PALM and STORM are slower than conventional live-imaging technologies and /or require high laser power, rendering these approaches unsuitable for super-resolved live imaging.

We will present actual membrane trafficking events captured using Andor’s Super-Resolution Radial Fluctuation (SRRF-stream) implemented via the highly sensitive iXon camera. Andor´s imaging technology offered new insights on the role of PI3-kinase in clathrin-mediated pinocytosis. Understanding this fundamental mechanism of vesicle trafficking could have implications for many diseases and might offer new therapeutic targets.

 

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