Subnanometer localization accuracy in widefield optical microscopy.
The common assumption that localization precision is the limit of localization accuracy and the typical absence of comprehensive calibration of optical microscopes present a widespread problem in localization microscopy. This combination causes overconfidence in measurement results with statistical uncertainties at the nanometer scale that are invalid due to systematic errors that can be orders of magnitude larger. In this article, we report a comprehensive solution to this underappreciated problem of false precision. We fabricate and measure arrays of subresolution apertures as reference materials, establishing new limits of measurement accuracy that approach the atomic scale over a submillimeter field. In a first application of our novel measurement capability, we introduce critical dimension localization microscopy, characterizing our fabrication accuracy and precision, and facilitating tests of nanofabrication processes and quality control of aperture arrays. In a second application, we apply these stable reference materials to answer open questions about the apparent instability of fluorescent nanoparticles that commonly serve as fiducial markers. By minimizing and quantifying systematic errors, we enable rigorous confidence in statistical precision as the limit of accuracy for localization microscopy at subnanometer scales.
Publisher URL: http://arxiv.org/abs/1710.09803
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