期刊
CHEMPHYSCHEM
卷 12, 期 3, 页码 609-626出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cphc.201000874
关键词
drug discovery; fluorescence lifetime imaging; FRET; high-throughput screening; proteins
资金
- UK Biotechnology and Biological Sciences Research Council (BBSRC) [BB/E003621/1, BB/H00713X/1]
- UK Engineering and Physical Sciences Research Council (EPSRC) [EP/F040202/1]
- UK Technology Strategy Board [CHBT/007/00030, EP/C54269X]
- Wellcome Trust [086114/ Z/08/Z]
- EPSRC
- Royal Society
- Wellcome Trust [086114/Z/08/Z] Funding Source: Wellcome Trust
- Biotechnology and Biological Sciences Research Council [BB/E003621/1, BB/H006095/1, BB/H00713X/1] Funding Source: researchfish
- Cancer Research UK [12011] Funding Source: researchfish
- Engineering and Physical Sciences Research Council [EP/F040202/1] Funding Source: researchfish
- BBSRC [BB/E003621/1, BB/H00713X/1, BB/H006095/1] Funding Source: UKRI
- EPSRC [EP/F040202/1] Funding Source: UKRI
A fluorescence lifetime imaging (FLIM) technology platform intended to read out changes in Forster 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.
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