Development of anisotropic super-resolution microscopy to examine SNARE function

Exocytosis, the process in which material is transported from the cell interior to the extracellular space, proceeds through a complex mechanism in which SNARE proteins play a key part. For the last decade, fluorescence microscopy greatly enhanced the understanding about the organisation and function of these macromolecules. However, conventional light microscopy is limited by diffraction and only allows studying specimens larger than 200 nm. In order to unravel the rapidly changing, nano-scale molecular architecture of the machinery driving exocytosis, which involves protein-protein and protein-lipid interactions in specialised sites in the cell, sophisticated methods need to be developed. This would allow us to see beyond the resolution limit. In my talk I will describe my work on the bulk organisation of SNAP-25 protein using confocal laser scanning microscopy and describe the development of a new super-resolution technique with the ability to detect and discriminate individual protein molecules in cells.