GENERALIZED EXTENSIONS OF CORRELATION SPECTROSCOPY

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The project involves extending the paradigm of spatio-temporal fluorescence correlation to include the calculation of pairwise correlation functions. Essentially this will allow measurement of pairwise correlation functions from two spatially separated pixels in an image time series. The analysis of pairwise correlation functions should prove useful for mapping membrane domains by detecting that barriers exist in the cell that hinder free diffusion. An appropriate membrane system (protein or lipid) and cell type will be selected for trial experiments. We will also explore other extensions of image correlation spectroscopy, specifically developing line kICS (reciprocal or k-space method) to separate binding and unbinding from transport for measurements of adhesion related macromolecules within the context of focal adhesions in living cells. Finally we will explore the measurement of FRET via a microscope equipped with APD detectors.