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FLIM FRET technology for drug discovery: automated multiwell-plate high-content analysis, multiplexed readouts and application in situ

Title: FLIM FRET technology for drug discovery: automated multiwell-plate high-content analysis, multiplexed readouts and application in situ
Authors: Kumar, S
Alibhai, D
Margineanu, A
Laine, R
Kennedy, G
McGinty, J
Warren, S
Kelly, D
Alexandrov, Y
Munro, I
Talbot, C
Stuckey, DW
Kimberly, C
Viellerobe, B
Lacombe, F
Lam, EW-F
Taylor, H
Dallman, MJ
Stamp, G
Murray, EJ
Stuhmeier, F
Sardini, A
Katan, M
Elson, DS
Neil, MAA
Dunsby, C
French, PMW
Item Type: Journal Article
Abstract: A fluorescence lifetime imaging (FLIM) technology platform intended to read out changes in Fçrster 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.
Issue Date: 17-Feb-2011
Date of Acceptance: 1-Feb-2011
URI: http://hdl.handle.net/10044/1/76638
DOI: 10.1002/cphc.201000874
ISSN: 1439-4235
Publisher: Wiley
Start Page: 609
End Page: 626
Journal / Book Title: ChemPhysChem: a European journal of chemical physics and physical chemistry
Volume: 12
Issue: 3
Copyright Statement: © 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim. 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
Sponsor/Funder: Biotechnology and Biological Sciences Research Council (BBSRC)
Engineering & Physical Science Research Council (EPSRC)
Biotechnology and Biological Sciences Research Council (BBSRC)
Cancer Research UK
Funder's Grant Number: BB/H00713X/1
EP/F040202/1
BB/E003621/1
12011
Keywords: Science & Technology
Physical Sciences
Chemistry, Physical
Physics, Atomic, Molecular & Chemical
Chemistry
Physics
drug discovery
fluorescence lifetime imaging
FRET
high-throughput screening
proteins
RESONANCE ENERGY-TRANSFER
LIFETIME IMAGING MICROSCOPY
VIRUS TYPE-1 GAG
OPTICAL PROJECTION TOMOGRAPHY
CYAN FLUORESCENT PROTEIN
FREQUENCY-DOMAIN
LIVING CELLS
LIVE CELLS
CORRELATION SPECTROSCOPY
SIGNALING EVENTS
Cell Line
Drug Evaluation, Preclinical
Fluorescence Resonance Energy Transfer
Fluorescent Dyes
Green Fluorescent Proteins
Humans
Microscopy, Fluorescence
Protein Binding
Proteins
Rhodamines
gag Gene Products, Human Immunodeficiency Virus
Cell Line
Humans
Rhodamines
Proteins
Green Fluorescent Proteins
Fluorescent Dyes
Microscopy, Fluorescence
Fluorescence Resonance Energy Transfer
Drug Evaluation, Preclinical
Protein Binding
gag Gene Products, Human Immunodeficiency Virus
Chemical Physics
0306 Physical Chemistry (incl. Structural)
0202 Atomic, Molecular, Nuclear, Particle and Plasma Physics
0307 Theoretical and Computational Chemistry
Edition: Forster Resonance Energy Transfer Issue
Publication Status: Published
Online Publication Date: 2011-02-17
Appears in Collections:Physics
Department of Surgery and Cancer
Photonics
Faculty of Natural Sciences