Two-photon MINFLUX with doubled localization precision

Kun Zhao; Xinzhu Xu; Wei Ren; Dayong Jin; Peng Xi

doi:10.1186/s43593-021-00011-x

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Two-photon MINFLUX with doubled localization precision

eLight Vol. 2, Issue 1, (2022)

LETTER | Updated：2024-09-28

- Two-photon MINFLUX with doubled localization precision
- Affiliations：
  
  Department of Biomedical Engineering, College of Future Technology, Peking University, Beijing 100871, China
  Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, 30332Georgia, USA
  UTS-SUStech Joint Research Centre for Biomedical Materials & Devices, Department of Biomedical Engineering, College of Engineering, Southern University of Science and Technology, Shenzhen, Guangdong 518055, China
  National Biomedical Imaging Center, Peking University, Beijing 100871, China
- Author bio：
  
  *xipeng@pku.edu.cn
- Funds：
- DOI：10.1186/s43593-021-00011-x
  CLC：
- Published：2022-12，
  
  Published Online：25 March 2022，
  
  Received：01 September 2021，
  
  Revised：10 December 2021，
  
  Accepted：22 December 2021
- Accepted：
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Kun Zhao, Xinzhu Xu, Wei Ren, et al. Two-photon MINFLUX with doubled localization precision. [J]. eLight, 2022,2(1).
DOI：

Kun Zhao, Xinzhu Xu, Wei Ren, et al. Two-photon MINFLUX with doubled localization precision. [J]. eLight, 2022,2(1). DOI： 10.1186/s43593-021-00011-x.

摘要

Abstract

Achieving localization with molecular precision has been of great interest for extending fluorescence microscopy to nanoscopy. MINFLUX pioneers this transition through point spread function (PSF) engineering

yet its performance is primarily limited by the signal-to-background ratio. Here we demonstrate theoretically that two-photon MINFLUX(2p-MINFLUX) could double its localization precision through PSF engineering by nonlinear effect. Cramér-Rao Bound(CRB) is studied as the maximum localization precision

and CRB of two-photon MINFLUX is halved compared to single-photon MINFLUX (1p-MINFLUX) in all three dimensions. Meanwhile

in order to achieve same localization precision with 1p-MINFLUX

2p-MINFLUX requires only 1/4 of fluorescence photons. Exploiting simultaneous two-photon excitation of multiple fluorophore species

2p-MINFLUX may have the potential for registration-free nanoscopy and multicolor tracking.

关键词

Keywords

Super-resolution microscopySingle-molecule localizationMINFLUXTwo-photon microscopy

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