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Cellular membranes have complex lipid and protein structures that are laterally organized for optimized molecular recognition and signal transduction processes. Knowledge of nanometer-scale lateral organization and its function is of great importance in the analysis of receptor-based signaling. In model membranes, we studied in detail the chemical and physical factors which result in lateral organization of lipids and lipid-mediated protein sequestration into signaling domains. In biological membranes, we mapped the location and follow the dynamic activity of specific membrane proteins involved in the immunological response of mast cells. These studies were enabled by our development of advanced imaging methods that provided both high spatial resolution and sensitivity to dynamical processes. Our technical approach was to combine the high sensitivity and time resolution of fluorescence imaging with the high lateral resolution of atomic force microscopy (AFM). Simultaneous fluorescence and AFM imaging allows correlation of the distribution and dynamic activity of specific biomolecules via fluorescence labeling with complete topographic information of the membrane. Overall, our unique imaging capabilities enabled us to examine membrane structure and function with much greater detail than was previously possible and thus provide a better understanding of cellular signaling

Studies of signaling domains in model and biological membranes through advanced imaging techniques: final report.