title: Time-resolved fluorescence in lipid bilayers: selected applications and advantages over steady state.
abstract: Fluorescence methods are versatile tools for obtaining dynamic and topological information about biomembranes because the molecular interactions taking place in lipid membranes frequently occur on the same timescale as fluorescence emission. The fluorescence intensity decay, in particular, is a powerful reporter of the molecular environment of a fluorophore. The fluorescence lifetime can be sensitive to the local polarity, hydration, viscosity, and/or presence of fluorescence quenchers/energy acceptors within several nanometers of the vicinity of a fluorophore. Illustrative examples of how time-resolved fluorescence measurements can provide more valuable and detailed information about a system than the time-integrated (steady-state) approach will be presented in this review: 1), determination of membrane polarity and mobility using time-dependent spectral shifts; 2), identification of submicroscopic domains by fluorescence lifetime imaging microscopy; 3), elucidation of membrane leakage mechanisms from dye self-quenching assays; and 4), evaluation of nanodomain sizes by time-resolved Förster resonance energy transfer measurements.
authors.affiliation: Department of Biophysical Chemistry, J. Heyrovský Institute of Physical Chemistry of the Academy of Sciences of the Czech Republic, v.v.i., Prague, Czech Republic.
authors.forename: Mariana
authors.initials: M
authors.lastname: Amaro
authors.affiliation: Department of Biophysical Chemistry, J. Heyrovský Institute of Physical Chemistry of the Academy of Sciences of the Czech Republic, v.v.i., Prague, Czech Republic.
authors.forename: Radek
authors.initials: R
authors.lastname: Šachl
authors.affiliation: Department of Biophysical Chemistry, J. Heyrovský Institute of Physical Chemistry of the Academy of Sciences of the Czech Republic, v.v.i., Prague, Czech Republic.
authors.forename: Piotr
authors.initials: P
authors.lastname: Jurkiewicz
authors.affiliation: Centre for Molecular Chemistry and Physics and Instituto de Nanociência e Nanotecnologia, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal; Departamento Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Lisbon, Portugal.
authors.forename: Ana
authors.initials: A
authors.lastname: Coutinho
authors.affiliation: Centre for Molecular Chemistry and Physics and Instituto de Nanociência e Nanotecnologia, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.
authors.forename: Manuel
authors.initials: M
authors.lastname: Prieto
authors.affiliation: Department of Biophysical Chemistry, J. Heyrovský Institute of Physical Chemistry of the Academy of Sciences of the Czech Republic, v.v.i., Prague, Czech Republic. Electronic address: martin.hof@jh-inst.cas.cz.
authors.forename: Martin
authors.initials: M
authors.lastname: Hof
mesh_terms: D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry*; D008856:Microscopy, Fluorescence / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D008051:Lipid Bilayers