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Proceedings Paper

FLIM, FRET and high content analysis
Author(s): Edwin Garcia; Wenjun Guo; Sunil Kumar; Frederik Görlitz; Hugh Sparks; Yuriy Alexandrov; Ian Munro; Douglas J. Kelly; Sean Warren; George Chennell; Alessandro Sardini; David Carling; Peter Thorpe; Christopher Dunsby; Paul M. W. French
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Paper Abstract

Fluorescence lifetime imaging (FLIM) provides a means to contrast different molecular species and to map variations in the local fluorophore molecular environment, including to read out Förster resonant energy transfer (FRET), e.g. to assay protein interactions or genetically expressed FRET biosensors. We have implemented wide-field time-gated FLIM in a modular open automated microscopy platform for high content analysis (HCA). To demonstrate its relevance to drug discovery, we have demonstrated the capability of our openFLIM HCA platform to assay interactions of low copy number endogenous proteins in yeast cells labelled with fluorescent proteins. We have also demonstrated the capability of multiwell plate FLIM assays to provide readouts of a FRET biosensor in 2-D and 3-D cell cultures.

Paper Details

Date Published: 2 March 2020
PDF: 8 pages
Proc. SPIE 11244, Multiphoton Microscopy in the Biomedical Sciences XX, 1124417 (2 March 2020); doi: 10.1117/12.2547517
Show Author Affiliations
Edwin Garcia, Imperial College London (United Kingdom)
Wenjun Guo, Imperial College London (United Kingdom)
The Francis Crick Institute (United Kingdom)
Sunil Kumar, Imperial College London (United Kingdom)
The Francis Crick Institute (United Kingdom)
Frederik Görlitz, Imperial College London (United Kingdom)
Hugh Sparks, Imperial College London (United Kingdom)
Yuriy Alexandrov, Imperial College London (United Kingdom)
The Francis Crick Institute (United States)
Ian Munro, Imperial College London (United Kingdom)
Douglas J. Kelly, Imperial College London (United Kingdom)
Sean Warren, Imperial College London (United Kingdom)
George Chennell, Imperial College London (United Kingdom)
Alessandro Sardini, Imperial College London (United Kingdom)
David Carling, Imperial College London (United Kingdom)
Peter Thorpe, Queen Mary Univ. of London (United Kingdom)
Christopher Dunsby, Imperial College London (United Kingdom)
Paul M. W. French, Imperial College London (United Kingdom)


Published in SPIE Proceedings Vol. 11244:
Multiphoton Microscopy in the Biomedical Sciences XX
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)

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