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Fast, robust and precise 3D localization for arbitrary point spread functions

By Yiming Li, Markus Mund, Philipp Hoess, Ulf Matti, Bianca Nijmeijer, Vilma Jimenez Sabinina, Jan Ellenberg, Ingmar Schoen, Jonas Ries

Posted 08 Aug 2017
bioRxiv DOI: 10.1101/172643 (published DOI: 10.1038/nmeth.4661)

We present a fitter for 3D single-molecule localization of arbitrary, experimental point spread functions (PSFs) that reaches minimum uncertainty for EMCCD and sCMOS cameras, and achieves more than 105 fits/s. We provide tools to robustly model experimental PSFs and correct for depth induced aberrations, which allowed us to achieve an unprecedented 3D resolution with engineered astigmatic PSFs, and acquire high quality 3D superresolution images even on standard microscopes without 3D optics.

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