A cellular screening assay using analysis of metal-modified fluorescence lifetime.

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.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Cade NI,Fruhwirth G,Archibald SJ,Ng T,Richards D

doi

10.1016/j.bpj.2010.03.016

subject

Has Abstract

pub_date

2010-06-02 00:00:00

pages

2752-7

issue

11

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(10)00346-2

journal_volume

98

pub_type

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