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  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., et al. (2022). Optimal precision and accuracy in 4Pi-STORM using dynamic spline PSF models. Nature Methods, 19(5), 603-612. doi:10.1038/s41592-022-01465-8.

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Item Permalink: https://hdl.handle.net/21.11116/0000-000A-8C1F-7 Version Permalink: https://hdl.handle.net/21.11116/0000-000A-8C20-4
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 Creators:
Bates, M., Author           
Keller-Findeisen, J., Author           
Przybylski, A., Author           
Hüper, A., Author
Stephan, T., Author           
Ilgen, P., Author           
Cereceda Delgado, A. R.1, Author           
D’Este, E.2, Author           
Egner, A., Author
Jakobs, S., Author           
Sahl, S. J., Author           
Hell, S. W.1, Author           
Affiliations:
1Optical Nanoscopy, Max Planck Institute for Medical Research, Max Planck Society, ou_2364730              
2Max Planck Institute for Medical Research, Max Planck Society, ou_1125545              

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Free keywords: Fluorescence imaging; Single-molecule biophysics; Super-resolution microscopy
 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 capa-bilities by investigating protein and nucleic acid organization in primary neurons and mammalian mitochondria.

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Language(s): eng - English
 Dates: 2022-05-162022-05
 Publication Status: Issued
 Pages: 27
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1038/s41592-022-01465-8
 Degree: -

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Title: Nature Methods
  Other : Nature Methods
Source Genre: Journal
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Publ. Info: New York, NY : Nature Publishing Group
Pages: - Volume / Issue: 19 (5) Sequence Number: - Start / End Page: 603 - 612 Identifier: ISSN: 1548-7091
CoNE: https://pure.mpg.de/cone/journals/resource/111088195279556