SGER: Multicolor Total Internal Reflection Fluorescence Microscopy

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0110070SimonThe primary goal of this proposal is to develop techniques to image two colors simultaneously at video rates (30 frames/sec) so that either two different probes, or a single probe with two different emissions, can be imaged synchronously. Further, to develop the techniques for using multiple lasers for excitation. This goal will allow two different kinds of studies: First, it would allow multiple proteins to be studied in a single cell. This is important for addressing questions of protein-protein interactions. Second, it will allow many physiological studies using dual-emission dyes that can be used to quantify important cell signaling molecules such as calcium or pH. The spatial and temporal dynamics of these molecules are key for understanding many aspects of cellular signaling.