Open AccessReal-time 3D single-molecule localization using experimental point spread functions
Author(s) -
Yiming Li,
Markus Mund,
Philipp Hoess,
Joran Deschamps,
Ulf Matti,
Bianca Nijmeijer,
Vilma Jimenez Sabinina,
Jan Ellenberg,
Ingmar Schoen,
Jonas Ries
Publication year - 2018
Publication title -
nature methods
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 19.469
H-Index - 318
eISSN - 1548-7105
pISSN - 1548-7091
DOI - 10.1038/nmeth.4661
Subject(s) - microscope , microscopy , point spread function , optical microscope , resolution (logic) , super resolution microscopy , optics , computer science , point (geometry) , superresolution , image quality , computer vision , artificial intelligence , biological system , image (mathematics) , algorithm , physics , mathematics , biology , scanning confocal electron microscopy , scanning electron microscope , geometry
We present a real-time fitter for 3D single-molecule localization microscopy using experimental point spread functions (PSFs) that achieves minimal uncertainty in 3D on any microscope and is compatible with any PSF engineering approach. We used this method to image cellular structures and attained unprecedented image quality for astigmatic PSFs. The fitter compensates for most optical aberrations and makes accurate 3D super-resolution microscopy broadly accessible, even on standard microscopes without dedicated 3D optics.