Real-time 3D single-molecule localization using experimental point spread functions

被引:189
作者
Li, Yiming [1 ]
Mund, Markus [1 ]
Hoess, Philipp [1 ]
Deschamps, Joran [1 ]
Matti, Ulf [1 ]
Nijmeijer, Bianca [1 ]
Sabinina, Vilma Jimenez [1 ]
Ellenberg, Jan [1 ]
Schoen, Ingmar [2 ]
Ries, Jonas [1 ]
机构
[1] European Mol Biol Lab, Cell Biol & Biophys Unit, Heidelberg, Germany
[2] Royal Coll Surgeons Ireland, Mol & Cellular Therapeut, Dublin, Ireland
基金
欧洲研究理事会;
关键词
OPTICAL RECONSTRUCTION MICROSCOPY; SUPERRESOLUTION MICROSCOPY; FLUORESCENCE MICROSCOPY; PARTICLE-TRACKING; RESOLUTION; ABERRATIONS; ALGORITHMS; NANOSCOPY; ACCURACY; PROBES;
D O I
10.1038/nmeth.4661
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:367 / +
页数:7
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