title: Single-image axial localization precision analysis for individual fluorophores.
abstract: Bio-mechanism investigations demand single particle tracking with high spatial and temporal resolutions which require single fluorophore 3D localization measurements with matching precision and speed. Although the precision for lateral-localization measurements is well described by an analytical expression, for the axial direction, it is often obtained by repeating location measurements or by estimating a lower bound. Here, we report a precision expression for an axial-localization method that analyzes the standard deviations of single fluorophores' intensity profiles. Like the lateral-localization precision, this expression includes all relevant experimental effects measurable from a gaussian intensity profile of the fluorophore. This expression completes the precision analysis for single-image 3D localization of individual fluorophores and lifts the temporal resolution to the typical exposure timescales of milliseconds.
authors.affiliation: Department of Physics, Washington University, St. Louis, MO 63130, USA.
authors.forename: Michael C
authors.initials: MC
authors.lastname: DeSantis
authors.forename: Shannon Kian
authors.initials: SK
authors.lastname: Zareh
authors.forename: Xianglu
authors.initials: X
authors.lastname: Li
authors.forename: Robert E
authors.initials: RE
authors.lastname: Blankenship
authors.forename: Y M
authors.initials: YM
authors.lastname: Wang
mesh_terms: D000465:Algorithms; D005456:Fluorescent Dyes / Q000032:analysis*; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes