title: Optimized localization analysis for single-molecule tracking and super-resolution microscopy.
abstract: We optimally localized isolated fluorescent beads and molecules imaged as diffraction-limited spots, determined the orientation of molecules and present reliable formulas for the precision of various localization methods. Both theory and experimental data showed that unweighted least-squares fitting of a Gaussian squanders one-third of the available information, a popular formula for its precision exaggerates beyond Fisher's information limit, and weighted least-squares may do worse, whereas maximum-likelihood fitting is practically optimal.
authors.affiliation: Department of Micro- and Nanotechnology, Technical University of Denmark, Kongens Lyngby, Denmark.
authors.forename: Kim I
authors.initials: KI
authors.lastname: Mortensen
authors.forename: L Stirling
authors.initials: LS
authors.lastname: Churchman
authors.forename: James A
authors.initials: JA
authors.lastname: Spudich
authors.forename: Henrik
authors.initials: H
authors.lastname: Flyvbjerg
mesh_terms: D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D016013:Likelihood Functions; D008856:Microscopy, Fluorescence / Q000379:methods*; D036103:Nanotechnology; D017785:Photons