Combined topographical and high speed confocal imaging systems for interdisciplinary membrane and cell biology research. (360G-Wellcome-094266_Z_10_Z)

This proposal is for 2 cutting-edge imaging systems, which will be applied individually or in combination to enable (i) topographical scanning of surface membrane structure in living cells or artificial membrane bilayers (ii) focal dispensing of small molecules (e.g. channel agonists) at the cell surface or within specialized membrane invaginations (iii) ion channel recordings from regions of interest identified by topographical scanning (iv) confocal imaging of fluorescently tagged membrane proteins, which can be correlated with topographical information (v) high speed full-frame confocal fluorescence imaging of intracellular ions or other components of signaling cascades. The first of these imaging systems is the Ionscope scanning ion conductance microscope (SICM), which can resolve the surface structure of living cells at nanometer resolution, without making contact with the sample. The second imaging system is the Andor Revolution confocal microscope. This is a high perf ormance spinning disk confocal system, capable of collecting full x-y images at hundreds of frames per second from a user-defined confocal plane. These systems will enable a step change in ongoing interdisciplinary projects that address, for example, basic research on membrane structure and function, disease mechanisms (e.g. amyloid diseases, cardiac hypertrophy) and novel delivery methods for therapeutic agents.