title: FRET-FLIM applications in plant systems.
abstract: A hallmark of cellular processes is the spatio-temporally regulated interplay of biochemical components. Assessing spatial information of molecular interactions within living cells is difficult using traditional biochemical methods. Developments in green fluorescent protein technology in combination with advances in fluorescence microscopy have revolutionised this field of research by providing the genetic tools to investigate the spatio-temporal dynamics of biomolecules in live cells. In particular, fluorescence lifetime imaging microscopy (FLIM) has become an inevitable technique for spatially resolving cellular processes and physical interactions of cellular components in real time based on the detection of Förster resonance energy transfer (FRET). In this review, we provide a theoretical background of FLIM as well as FRET-FLIM analysis. Furthermore, we show two cases in which advanced microscopy applications revealed many new insights of cellular processes in living plant cells as well as in whole plants.
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich, NR4 7UH, UK.
authors.forename: Christoph A
authors.initials: CA
authors.lastname: Bücherl
authors.forename: Arjen
authors.initials: A
authors.lastname: Bader
authors.forename: Adrie H
authors.initials: AH
authors.lastname: Westphal
authors.forename: Sergey P
authors.initials: SP
authors.lastname: Laptenok
authors.forename: Jan Willem
authors.initials: JW
authors.lastname: Borst
mesh_terms: D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000737:chemistry*; D008856:Microscopy, Fluorescence / Q000379:methods*; D061848:Optical Imaging; D059828:Plant Cells / Q000737:chemistry / Q000648:ultrastructure; D010944:Plants / Q000737:chemistry* / Q000648:ultrastructure
chemical_list: D049452:Green Fluorescent Proteins