A Cellular Screening Assay Using Analysis of Metal-Modified Fluorescence Lifetime
A Cellular Screening Assay Using Analysis of Metal-Modified Fluorescence Lifetime
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DOI：
10.1016/j.bpj.2010.03.016
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发表时间：
2010-06-02
影响因子：
3.4
通讯作者：
Richards, David
中科院分区：
文献类型：
--
作者：
Cade, Nicholas I.;Fruhwirth, Gilbert;Richards, David
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.
