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Proceedings Paper

A cellular assay using metal-modified fluorescence lifetime analysis for high-content screening of protein internalization
Author(s): Nic Cade; Gilbert Fruhwirth; Stephen J. Archibald; Tony Ng; David Richards
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Paper Abstract

Current high-content screening (HCS) techniques involve the analysis of cellular assays using high-resolution imaging combined with sophisticated algorithms for automated image analysis. Commercially available platforms are invariably highly specialised and expensive. Here we present a novel assay utilising changes in fluorescence lifetime in the vicinity of a rough Au film. A mammary carcinoma cell line was created expressing EGFP in the membrane, and cells were plated onto multi-well slides covered with a 30 nm Au film. FLIM images show a large reduction in lifetime for membrane-bound GFP in close proximity to the Au surface. Addition of a suitable ligand leads to internalization of the GFP with a corresponding increase in lifetime. The degree of internalization can be very quickly and easily checked using standard lifetime analysis techniques, with no need for image analysis. We demonstrate the method by comparing the efficacies of two small molecule inhibitors interfering with the internalization process.

Paper Details

Date Published: 17 May 2010
PDF: 8 pages
Proc. SPIE 7715, Biophotonics: Photonic Solutions for Better Health Care II, 77150P (17 May 2010); doi: 10.1117/12.852547
Show Author Affiliations
Nic Cade, King's College London (United Kingdom)
Gilbert Fruhwirth, King's College London (United Kingdom)
Stephen J. Archibald, The Univ. of Hull (United Kingdom)
Tony Ng, King's College London (United Kingdom)
David Richards, King's College London (United Kingdom)

Published in SPIE Proceedings Vol. 7715:
Biophotonics: Photonic Solutions for Better Health Care II
Jürgen Popp; Wolfgang Drexler; Valery V. Tuchin; Dennis L. Matthews, Editor(s)

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