Detection of nucleic acid–protein interactions in leaves using fluorescence lifetime imaging microscopy ($)

FRET-FLIM (Fluorescence Resonance Energy Transfer – Fluorescent Lifetime Imaging Microscopy) is a method that allows detection of interactions between two fluorescently-tagged molecules. Camborde and Jauneau et al. describe the use of FRET-FLIM to identify interactions in leaves between GFP-tagged DNA-binding proteins and nucleic acids (DNA or RNA), labeled with Sytox Orange. When the two molecules are in proximity, energy is transferred from GFP to Sytox Orange, which decreases the fluorescence lifetime of the GFP. Like other articles in Nature Protocols, this article provides a very detailed method with plenty of hints for troubleshooting. Nature Protocols 10.1038/nprot.2017.076