Optimal precision and accuracy in 4Pi-STORM using dynamic spline PSF models

2022 | journal article; research paper. A publication with affiliation to the University of Göttingen.

Jump to:Cite & Linked | Documents & Media | Details | Version history

Cite this publication

​Optimal precision and accuracy in 4Pi-STORM using dynamic spline PSF models​
Bates, M.; Keller-Findeisen, J.; Przybylski, A.; Hüper, A.; Stephan, T.; Ilgen, P. & Cereceda Delgado, A. R. et al.​ (2022) 
Nature Methods19(5) pp. 603​-612​.​ DOI: https://doi.org/10.1038/s41592-022-01465-8 

Documents & Media

s41592-022-01465-8.pdf16.15 MBAdobe PDF

License

Published Version

Attribution 4.0 CC BY 4.0

Details

Authors
Bates, Mark; Keller-Findeisen, Jan; Przybylski, Adrian; Hüper, Andreas; Stephan, Till; Ilgen, Peter; Cereceda Delgado, Angel R.; D’Este, Elisa; Egner, Alexander; Jakobs, Stefan ; Hell, Stefan W. 
Abstract
Abstract Coherent fluorescence imaging with two objective lenses (4Pi detection) enables single-molecule localization microscopy with sub-10 nm spatial resolution in three dimensions. Despite its outstanding sensitivity, wider application of this technique has been hindered by complex instrumentation and the challenging nature of the data analysis. Here we report the development of a 4Pi-STORM microscope, which obtains optimal resolution and accuracy by modeling the 4Pi point spread function (PSF) dynamically while also using a simpler optical design. Dynamic spline PSF models incorporate fluctuations in the modulation phase of the experimentally determined PSF, capturing the temporal evolution of the optical system. Our method reaches the theoretical limits for precision and minimizes phase-wrapping artifacts by making full use of the information content of the data. 4Pi-STORM achieves a near-isotropic three-dimensional localization precision of 2–3 nm, and we demonstrate its capabilities by investigating protein and nucleic acid organization in primary neurons and mammalian mitochondria.
Issue Date
2022
Journal
Nature Methods 
Project
SFB 1190: Transportmaschinen und Kontaktstellen zellulärer Kompartimente 
SFB 1190 | P01: Untersuchung der Unterschiede in der Zusammensetzung, Funktion und Position von individuellen MICOS Komplexen in einzelnen Säugerzellen 
SFB 1286: Quantitative Synaptologie 
SFB 1286 | A07: Der Aufbau des synaptischen Cytoskeletts 
Working Group
RG Jakobs (Structure and Dynamics of Mitochondria) 
RG Hell 
External URL
https://sfb1190.med.uni-goettingen.de/production/literature/publications/175
https://sfb1286.uni-goettingen.de/literature/publications/166
ISSN
1548-7091
eISSN
1548-7105
Language
English
Sponsor
European Molecular Biology Organization
Max-Planck-Gesellschaft
Deutsche Forschungsgemeinschaft

Reference

Citations


Social Media