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FLIM FRET Technology for Drug Discovery: Automated Multiwell‐Plate High‐Content Analysis, Multiplexed Readouts and Application in Situ
Author(s) -
Kumar Sunil,
Alibhai Dominic,
Margineanu Anca,
Laine Romain,
Kennedy Gordon,
McGinty James,
Warren Sean,
Kelly Douglas,
Alexandrov Yuriy,
Munro Ian,
Talbot Clifford,
Stuckey Daniel W.,
Kimberly Christopher,
Viellerobe Bertrand,
Lacombe Francois,
Lam Eric W.F.,
Taylor Harriet,
Dallman Margaret J.,
Stamp Gordon,
Murray Edward J.,
Stuhmeier Frank,
Sardini Alessandro,
Katan Matilda,
Elson Daniel S.,
Neil Mark A. A.,
Dunsby Chris,
French Paul M. W.
Publication year - 2011
Publication title -
chemphyschem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.016
H-Index - 140
eISSN - 1439-7641
pISSN - 1439-4235
DOI - 10.1002/cphc.201000874
Subject(s) - förster resonance energy transfer , fluorescence lifetime imaging microscopy , high content screening , confocal , confocal microscopy , microscopy , drug discovery , chemistry , biophysics , fluorescence , nanotechnology , materials science , cell , biology , physics , microbiology and biotechnology , biochemistry , optics
A fluorescence lifetime imaging (FLIM) technology platform intended to read out changes in Förster resonance energy transfer (FRET) efficiency is presented for the study of protein interactions across the drug‐discovery pipeline. FLIM provides a robust, inherently ratiometric imaging modality for drug discovery that could allow the same sensor constructs to be translated from automated cell‐based assays through small transparent organisms such as zebrafish to mammals. To this end, an automated FLIM multiwell‐plate reader is described for high content analysis of fixed and live cells, tomographic FLIM in zebrafish and FLIM FRET of live cells via confocal endomicroscopy. For cell‐based assays, an exemplar application reading out protein aggregation using FLIM FRET is presented, and the potential for multiple simultaneous FLIM (FRET) readouts in microscopy is illustrated.