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Proceedings Paper

A high-content screening platform utilizing polarization anisotropy and FLIM microscopy
Author(s): Daniel R. Matthews; Simon M. Ameer-Beg; Paul Barber; Glenn P. Pierce; Robert G. Newman; Boris Vojnovic; Leo M. Carlin; Melanie D. Keppler; Tony Ng; Klaus Suhling; Malcolm Irving
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Paper Abstract

An automated high-content screening microscope has been developed which uses fluorescence anisotropy imaging and fluorescence lifetime microscopy to identify Förster resonant energy transfer between eGFP and mRPF1 in drug screening assays. A wide-field polarization resolved imager is used to simultaneously capture the parallel and perpendicular components of both eGFP and mRFP1 fluorescence emission to provide a high-speed measurement of acceptor depolarization. Donor excited state lifetime measurements performed using laser scanning microscopy is then used to determine the FRET efficiency in a particular assay. A proof-of-principle assay is performed using mutant Jurkat human T-cells to illustrate the process by which FRET is first identified and then quantified by our high-content screening system.

Paper Details

Date Published: 5 March 2008
PDF: 12 pages
Proc. SPIE 6859, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VI, 685919 (5 March 2008); doi: 10.1117/12.763310
Show Author Affiliations
Daniel R. Matthews, King's College London (United Kingdom)
Simon M. Ameer-Beg, King's College London (United Kingdom)
Paul Barber, Univ. of Oxford (United Kingdom)
Glenn P. Pierce, Univ. of Oxford (United Kingdom)
Robert G. Newman, Univ. of Oxford (United Kingdom)
Boris Vojnovic, Univ. of Oxford (United Kingdom)
Leo M. Carlin, King's College London (United Kingdom)
Melanie D. Keppler, King's College London (United Kingdom)
Tony Ng, King's College London (United Kingdom)
Klaus Suhling, King's College London (United Kingdom)
Malcolm Irving, King's College London (United Kingdom)


Published in SPIE Proceedings Vol. 6859:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VI
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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