title: Quantifying cellular dynamics by fluorescence resonance energy transfer (FRET) microscopy.
abstract: The cell is a spatially organized system whose function emerges from the complex interaction of molecular components. Such local interaction of nanometer-sized molecules generates patterns that span throughout the cell. Those patterns, in turn, regulate the molecular interactions. Understanding such simultaneous bidirectional causation requires quantifying the spatio-temporal progression of biochemical reactions in the context of a living cell. Due to its ability to resolve micrometer-sized structures, biological microscopy has been instrumental to the discovery and understanding of living systems. Functional fluorescence microscopy allows a cellular dynamic topographic map of proteins to be overlaid with topological information on the causality that determines protein state. Here we describe how Förster/fluorescence resonance energy transfer (FRET) can be used to measure the phosphorylation state of proteins in the context of the cell.
authors.affiliation: Department of Systemic Cell Biology, Max Planck Institute for Molecular Physiology, Dortmund, Germany.
authors.forename: Hernán E
authors.initials: HE
authors.lastname: Grecco
authors.forename: Philippe I H
authors.initials: PI
authors.lastname: Bastiaens
mesh_terms: D000818:Animals; D002232:Carbocyanines / Q000378:metabolism; D002460:Cell Line; D002468:Cell Physiological Phenomena / Q000502:physiology*; D031541:Fluorescence Resonance Energy Transfer; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence; D014162:Transfection
chemical_list: D002232:Carbocyanines; C089187:cyanine dye 3; D049452:Green Fluorescent Proteins