Discussion

Although fluorescence lifetime imaging (FLIM) is established in life sciences applications as a method whereby quantitative information may be extracted from time-resolved fluorescence signals, FLIM has not been widely adopted in an HCA context. We have developed an automated plate reading microscope that combines FLIM with multispectral, bright field and phase contrast imaging, typically acquiring multimodal image data without supervision from more than 120 fields of view in an hour. This instrument is being developed as open hardware with open source implementations of the software for data acquisition, analysis and management. We present an overview of the instrument and its application to assay protein interactions using Förster resonance energy transfer and to label-free HCA assays using cellular autofluorescence lifetime to investigate the effects of metabolic modulators in live cells.