A Cellular Screening Assay Using Analysis of Metal-Modified Fluorescence Lifetime |
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Authors: | Nicholas I. Cade Gilbert Fruhwirth Tony Ng |
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Affiliation: | † Department of Physics, King's College London, London, United Kingdom ‡ Division of Cancer Studies and Randall Division of Cell and Molecular Biophysics, King's College London, London, United Kingdom § Department of Chemistry, The University of Hull, Hull, United Kingdom |
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Abstract: | Current methods for screening cell receptor internalization often require complex image analysis with limited sensitivity. Here we describe a novel bioassay based on detection of changes in global fluorescence lifetime above a gold substrate, with superresolution axial sensitivity and no need for image analysis. We show that the lifetime of enhanced green fluorescent protein expressed in a cellular membrane is greatly reduced in close proximity to the gold, resulting in a distance-dependent lifetime distribution throughout the cell. We demonstrate the application of this phenomenon in a screening assay by comparing the efficacies of two small molecule inhibitors interfering with the internalization process of a G protein-coupled receptor. |
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