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 Jjournal_title
Biophysical journalauthors
Cade NI,Fruhwirth G,Archibald SJ,Ng T,Richards Ddoi
10.1016/j.bpj.2010.03.016subject
Has Abstractpub_date
2010-06-02 00:00:00pages
2752-7issue
11eissn
0006-3495issn
1542-0086pii
S0006-3495(10)00346-2journal_volume
98pub_type
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