A Cellular Screening Assay Using Analysis of Metal-Modified Fluorescence Lifetime

作者:Cade Nicholas I*; Fruhwirth Gilbert; Archibald Stephen J; Ng Tony; Richards David
来源:Biophysical Journal, 2010, 98(11): 2752-2757.
DOI:10.1016/j.bpj.2010.03.016

摘要

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.

  • 出版日期2010-6-2