Development of the next generation of sicm for live cell imaging

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The cell surface is a complex arrangement of many molecules and this arrangement changes in time for the cell to perform specific functions and in response to stimuli. At present we can only resolve the molecular structure on frozen cells, so dynamics cannot be followed or we can follow just one component on the surface of live cells by the use of labelling. One method that has been developed to probe live cells at sufficient resolution to determine proteins on the surface is based on a scanned nanopipette. This controlled just above the soft cell surface so that it never touches and can be rastered over the surface to determine the topography. We aim to build on this method so that it can image more complex cell surfaces faster and can also locally apply reagents to identfy the features that we detect. This will be done with sufficient resolution to detect indvidual protein complexes on the cell surface, their organization and how this changes with time. This will allow many new details of how the cell works at the nanoscale to be observed for the first time and opening up many new types of experiments