Nicholas I. Cade
A cellular screening assay using analysis of metal-modified fluorescence lifetime
Cade, Nicholas I.; Fruhwirth, Gilbert; Archibald, Stephen J.; Ng, Tony; Richards, David
Professor Steve Archibald S.J.Archibald@hull.ac.uk
Professor in Molecular Imaging
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.
Cade, N. I., Fruhwirth, G., Archibald, S. J., Ng, T., & Richards, D. (2010). A cellular screening assay using analysis of metal-modified fluorescence lifetime. Biophysical journal, 98(11), 2752-2757. https://doi.org/10.1016/j.bpj.2010.03.016
|Journal Article Type||Article|
|Acceptance Date||Jun 2, 2010|
|Publication Date||Jun 2, 2010|
|Peer Reviewed||Peer Reviewed|
|Keywords||CXCR4 antagonist, chemokine receptor, medicinal inorganic, cellular assay, FRET, fluorescence microscopy|
This file is under embargo due to copyright reasons.