Digital Imaging Microscope

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This proposal requests support for a multidisciplinary effort to develop the digital imaging microscope for the anlaysis and control of changes in molecular distribution underlying cell function. We propose to enhance the spatial and temporal resolution of fluorescence imaging by developing new microscope and camera systems for imaging in the millisecond domain and by developing computational methods for further extending resolution. We will develop methodology for quantitative analysis of 2-D, 3-D and 3-D time series fluorescence images and for determining error bounds on estimates of the concentration of a given molecule in a compartment. We will provide a powerful capability for the measurement and analysis of images by integrating tools for image display and interaction with automated feature extraction algorithms. We will develop tools to investigate the relationship of two or more molecules with respect to one another. We will incrased the algorithm computation speed to make the digital imaging microscope into a real- time device that allows interactive or automated control of experiments. Finally, we will add to the instrument the ability to control and modify molecular distribution in living cells by controlled photolysis of "caged" compunds.