title: Measuring image resolution in optical nanoscopy.
abstract: Resolution in optical nanoscopy (or super-resolution microscopy) depends on the localization uncertainty and density of single fluorescent labels and on the sample's spatial structure. Currently there is no integral, practical resolution measure that accounts for all factors. We introduce a measure based on Fourier ring correlation (FRC) that can be computed directly from an image. We demonstrate its validity and benefits on two-dimensional (2D) and 3D localization microscopy images of tubulin and actin filaments. Our FRC resolution method makes it possible to compare achieved resolutions in images taken with different nanoscopy methods, to optimize and rank different emitter localization and labeling strategies, to define a stopping criterion for data acquisition, to describe image anisotropy and heterogeneity, and even to estimate the average number of localizations per emitter. Our findings challenge the current focus on obtaining the best localization precision, showing instead how the best image resolution can be achieved as fast as possible.
authors.affiliation: Quantitative Imaging Group, Delft University of Technology, Delft, The Netherlands.
authors.forename: Robert P J
authors.initials: RP
authors.lastname: Nieuwenhuizen
authors.forename: Keith A
authors.initials: KA
authors.lastname: Lidke
authors.forename: Mark
authors.initials: M
authors.lastname: Bates
authors.forename: Daniela Leyton
authors.initials: DL
authors.lastname: Puig
authors.forename: David
authors.initials: D
authors.lastname: Grünwald
authors.forename: Sjoerd
authors.initials: S
authors.lastname: Stallinga
authors.forename: Bernd
authors.initials: B
authors.lastname: Rieger
mesh_terms: D005454:Fluorescence Polarization; D005456:Fluorescent Dyes; D021621:Imaging, Three-Dimensional; D008856:Microscopy, Fluorescence / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes