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.

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

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