title: Precisely and accurately localizing single emitters in fluorescence microscopy.
abstract: Methods based on single-molecule localization and photophysics have brought nanoscale imaging with visible light into reach. This has enabled single-particle tracking applications for studying the dynamics of molecules and nanoparticles and contributed to the recent revolution in super-resolution localization microscopy techniques. Crucial to the optimization of such methods are the precision and accuracy with which single fluorophores and nanoparticles can be localized. We present a lucid synthesis of the developments on this localization precision and accuracy and their practical implications in order to guide the increasing number of researchers using single-particle tracking and super-resolution localization microscopy.
authors.affiliation: 1] Laboratory of General Biochemistry and Physical Pharmacy, Ghent University, Ghent, Belgium. [2] Center for Nano- and Biophotonics, Ghent University, Ghent, Belgium.
authors.forename: Hendrik
authors.initials: H
authors.lastname: Deschout
authors.affiliation: 1] Department of Nanophysics, Italian Institute of Technology, Genoa, Italy. [2] Department of Physics, University of Genoa, Genoa, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Cella Zanacchi
authors.affiliation: Department of Physics and Astronomy, University of Maine, Orono, Maine, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Mlodzianoski
authors.affiliation: 1] Department of Nanophysics, Italian Institute of Technology, Genoa, Italy. [2] Department of Physics, University of Genoa, Genoa, Italy.
authors.forename: Alberto
authors.identifier: 0000000249165928
authors.initials: A
authors.lastname: Diaspro
authors.affiliation: 1] Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut, USA. [2] Department of Biomedical Engineering, Yale University, New Haven, Connecticut, USA.
authors.forename: Joerg
authors.initials: J
authors.lastname: Bewersdorf
authors.affiliation: Department of Physics and Astronomy, University of Maine, Orono, Maine, USA.
authors.forename: Samuel T
authors.initials: ST
authors.lastname: Hess
authors.affiliation: 1] Laboratory of General Biochemistry and Physical Pharmacy, Ghent University, Ghent, Belgium. [2] Center for Nano- and Biophotonics, Ghent University, Ghent, Belgium.
authors.forename: Kevin
authors.initials: K
authors.lastname: Braeckmans
mesh_terms: D005456:Fluorescent Dyes / Q000737:chemistry*; D008856:Microscopy, Fluorescence / Q000379:methods* / Q000592:standards*; D015203:Reproducibility of Results
chemical_list: D005456:Fluorescent Dyes