Live-Cell STED Microscope

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A mechanistic understanding of physiological processes necessitates information on structure-function relationships. At the molecular level, this objective is obstructed by the limited resolving power of the primary current imaging approaches: diffraction limited spatial resolution of conventional light microscopy and poor temporal resolution provided by electron microscopy. The recent development of super-resolution microscopy (SRM) methods has produced innovative solutions to bypass the diffraction barrier (Nobel Prize in Chemistry, 2014). At present, however, we face the main challenge of transferring these techniques to diverse physiological applications. The present proposal aims to address this bottleneck by engaging SRM by Live-Cell STED (stimulated emission depletion). The requested instrument configuration combines high spatial resolution (<100x100x100 nm in 3D, <30x30 nm in 2D) with the molecular specificity and live-imaging capacity of fluorescence microscopy. We have assembled a team of researchers representing the Faculties of Medicine and Life Sciences at Leipzig University who are experienced in analysing structure-function relationships of the nervous system at different levels of biological organisation. This will enable us to optimise the microscope for studies of nanoscopic sub-cellular structures and real-time protein dynamics in a wide range of neurophysiological applications. We anticipate that this strategy will help to deliver new information on fundamental principles of brain function.