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. 2017 May 3;8:15115. doi: 10.1038/ncomms15115

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Copyright © 2017, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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Simulated images of a diffusing fluorophore with diffusion constant 1 μm² s⁻¹, 1 photon per pixel background, EMCCD noise and varying localization uncertainty due to varying (a) spot amplitude N, (b,c) average defocus z and exposure time t_E. Image size 20-by-20 pixels, pixel size 80 nm, colorbar indicates photons per pixel.