Measuring Image Resolution in Optical Nanoscopy

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2013 Apr 30

PMID 23624665

Citations 296

Authors

Robert P J Nieuwenhuizen

Keith A Lidke

Mark Bates

Daniela Leyton Puig

David Grunwald

Sjoerd Stallinga

Bernd Rieger

Affiliations

Soon will be listed here.

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.

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