CAREER: A Computational and Analytical Approach to Single-Molecule Fluorescence Imaging for the Quantitative Analysis of Protein Interactions in Living Cells.

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In this CAREER project the PI will study pre-clustering of the IgE receptor system. The proposed approach is to use the intermittent and independent fluctuations of fluorescent probes that are either photo-switchable or naturally intermittent to extract individual trajectories even when there are clusters of fluorophores with sizes below the diffraction limit. The approach is based on finding parameters and states in hidden Markov models and this approach will be further developed. The specific aims of this project are (1) to use fluorescent probes that are either photo-switchable or naturally intermittent to find single membrane protein trajectories of high density membrane components; (2) to use the trajectories of closely spaced proteins in conjunction with physical models to extract the binding kinetics between single molecules and to map out the energy landscape of the cell membrane; (3) using the developed techniques, to characterize the behavior of the IgE binding receptor of RBL-2H3 cells. The project will provide multidisciplinary training for a graduate student as well as undergraduate students and offer annual, paid research mentorships to high school students in the Albuquerque, New Mexico area. A student exchange program with the Quantitative Imaging group at Delft Technical University, Netherlands will allow up to 3 months annual international study for graduate students or promising undergraduates. An upper level Biophysics course will be developed based on the research interests of the PI's department. Speakers will be invited from within the U.S. to give talks on related research in a biophysics seminar series.