High speed unsupervised fluorescence lifetime imaging confocal multiwell plate reader for high content analysis

We report an automated optically sectioning fluorescence lifetime imaging (FLIM) multiwell plate reader for high content analysis (HCA) in drug discovery and accelerated research in cell biology. The system utilizes a Nipkow disc confocal microscope and performs, unsupervised FLIM, with autofocus, automatic setting of acquisition parameters and automated localisation of cells in the field of view. We demonstrate its applications to test dye solutions, fixed and live cells and FLIM-FRET.A fluorescence lifetime map showing automatically acquired data from a cell where the fluorophores are undergoing FRET. The acquisition time including stage movement, auto-find and auto-exposure was similar to 20 s.[GRAPHICS](c) 2008 by WILEY-VCH Velag GmbH & Co. KGaA. Weinheim