Simple and robust 3D MINFLUX excitation with a variable phase plate

Takahiro Deguchi; Jonas Ries

doi:10.1038/s41377-024-01487-1

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Simple and robust 3D MINFLUX excitation with a variable phase plate

Original Articles

- Simple and robust 3D MINFLUX excitation with a variable phase plate
- Light: Science & Applications Vol. 13, Issue 7, Pages: 1354-1363(2024)
- Affiliations：
  
  1.European Molecular Biology Laboratory, Cell Biology and Biophysics, Heidelberg, Germany
  2.Max Perutz Labs, Vienna Biocenter Campus (VBC), Vienna, Austria
  3.University of Vienna, Center for Molecular Biology, Department of Structural and Computational Biology, Vienna, Austria
  4.University of Vienna, Faculty of Physics, Vienna, Austria
- Author bio：
  
  Jonas Ries (Jonas.ries@univie.ac.at)
- Funds：
- DOI：10.1038/s41377-024-01487-1
  CLC：
- Received：28 January 2024，
  
  Revised：06 May 2024，
  
  Accepted：2024-05-16，
  
  Published Online：07 June 2024，
  
  Published：31 July 2024
- Accepted：
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Deguchi, T. & Ries, J. Simple and robust 3D MINFLUX excitation with a variable phase plate. Light: Science & Applications, 13, 1354-1363 (2024).
DOI：

Deguchi, T. & Ries, J. Simple and robust 3D MINFLUX excitation with a variable phase plate. Light: Science & Applications, 13, 1354-1363 (2024). DOI： 10.1038/s41377-024-01487-1.

摘要

Abstract

MINFLUX has achieved extraordinary resolution in superresolution imaging and single fluorophore tracking. It is based on localizing single fluorophores by rapid probing with a patterned beam that features a local intensity minimum. Current implementations

however

are complex and expensive and are limited in speed and robustness. Here

we show that a combination of an electro-optical modulator with a segmented birefringent element such as a spatial light modulator produces a variable phase plate for which the phase can be scanned on the MHz timescale. Bisected or top-hat phase patterns generate high-contrast compact excitation point-spread functions for MINFLUX localization in the x

and z-direction

respectively

which can be scanned across a fluorophore within a microsecond

switched within 60 microseconds and alternated among different excitation wavelengths. We discuss how to compensate for non-optimal performance of the components and present a robust 3D and multi-color MINFLUX excitation module

which we envision as an integral component of a high-performance and cost-effective open-source MINFLUX.

关键词

Keywords

references

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