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.