5 March 2008 A high-content screening platform utilizing polarization anisotropy and FLIM microscopy
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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.
© (2008) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Daniel R. Matthews, Daniel R. Matthews, Simon M. Ameer-Beg, Simon M. Ameer-Beg, Paul Barber, Paul Barber, Glenn P. Pierce, Glenn P. Pierce, Robert G. Newman, Robert G. Newman, Boris Vojnovic, Boris Vojnovic, Leo M. Carlin, Leo M. Carlin, Melanie D. Keppler, Melanie D. Keppler, Tony Ng, Tony Ng, Klaus Suhling, Klaus Suhling, Malcolm Irving, Malcolm Irving, } "A high-content screening platform utilizing polarization anisotropy and FLIM microscopy", Proc. SPIE 6859, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VI, 685919 (5 March 2008); doi: 10.1117/12.763310; https://doi.org/10.1117/12.763310

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