FLIM FRET Technology for Drug Discovery: Automated Multiwell-Plate High-Content Analysis, Multiplexed Readouts and Application in Situ
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 dr...
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Published in | Chemphyschem Vol. 12; no. 3; pp. 609 - 626 |
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Main Authors | , , , , , , , , , , , , , , , , , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Weinheim
WILEY-VCH Verlag
25.02.2011
WILEY‐VCH Verlag Wiley |
Subjects | |
Online Access | Get full text |
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Summary: | 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.
Chance of a lifetime: A fluorescence lifetime imaging (FLIM) technology platform to read out changes in Förster resonance energy transfer (FRET) efficiency is used for the study of protein interactions across the drug‐discovery pipeline. An automated FLIM multiwell‐plate reader acquires lifetime images of fixed and live cells labelled with fluorescent proteins in a few seconds per field of view (see picture) and can read a 96‐well plate in less than 15 min. |
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Bibliography: | ark:/67375/WNG-BGCFNGWR-D FLIM=Fluorescence Lifetime Imaging; FRET=Förster Resonance Energy Transfer Funded Access istex:BC2A0D08F5C5D8B17EF04ECBC17C0B4593CF12B8 BBSRC - No. BB/E003621/1; No. BB/H00713X/1 ArticleID:CPHC201000874 EPSRC - No. EP/F040202/1 Wellcome Trust - No. 086114/Z/08/Z UK Technology Strategy Board - No. CHBT/007/00030; No. EP/C54269X Re-use of this article is permitted in accordance with the Terms and Conditions set out at http://onlinelibrary.wiley.com/journal/10.1002/(ISSN) 1521-3773/homepage/2002_onlineopen.html |
ISSN: | 1439-4235 1439-7641 |
DOI: | 10.1002/cphc.201000874 |