Tetra-color superresolution microscopy based on excitation spectral demixing

Wanyan Wu; Shihang Luo; Chunyan Fan; Tianjie Yang; Shuwen Zhang; Wenxiang Meng; Tao Xu; Wei Ji; Lusheng Gu

doi:10.1038/s41377-022-01054-6

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Tetra-color superresolution microscopy based on excitation spectral demixing

Original Articles

- Tetra-color superresolution microscopy based on excitation spectral demixing
- Light: Science & Applications Vol. 12, Issue 1, Pages: 99-108(2023)
- Affiliations：
  
  1.Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China
  2.College of Life Science, University of Chinese Academy of Sciences, Beijing, 100049, China
  3.State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Innovation Academy for Seed Design, Chinese Academy of Sciences, Beijing, 100101, China
  4.School of Future Technology, University of Chinese Academy of Sciences, Beijing, 100049, China
  5.College of Advanced Agricultural Sciences, University of Chinese Academy of Sciences, Beijing, 100049, China
  6.Bioland Laboratory, Guangzhou, 510005, China
  7.Guangzhou Laboratory, Guangzhou, 510030, China
- Author bio：
  
  Tao Xu (xutao@ibp.ac.cn)
  Wei Ji (jiwei@ibp.ac.cn)
  Lusheng Gu (gulusheng@ibp.ac.cn)
- Funds：
- DOI：10.1038/s41377-022-01054-6
  CLC：
- Published：31 January 2023，
  
  Published Online：2 January 2023，
  
  Received：25 July 2022，
  
  Revised：29 November 2022，
  
  Accepted：2 December 2022
Scan for full text
Wu, W. Y. et al. Tetra-color superresolution microscopy based on excitation spectral demixing. Light: Science & Applications, 12, 99-108 (2023).
DOI：

Wu, W. Y. et al. Tetra-color superresolution microscopy based on excitation spectral demixing. Light: Science & Applications, 12, 99-108 (2023). DOI： 10.1038/s41377-022-01054-6.

摘要

Abstract

Multicolor imaging allows protein colocalizations and organelle interactions to be studied in biological research

which is especially important for single-molecule localization microscopy (SMLM). Here

we propose a multicolor method called excitation-resolved stochastic optical reconstruction microscopy (ExR-STORM). The method

which is based on the excitation spectrum of fluorescent dyes

successfully separated four spectrally very close far-red organic fluorophores utilizing three excitation lasers with cross-talk of less than 3%. Dyes that are only 5 nm apart in the emission spectrum were resolved

resulting in negligible chromatic aberrations. This method was extended to three-dimensional (3D) imaging by combining the astigmatic method

providing a powerful tool for resolving 3D morphologies at the nanoscale.

关键词

Keywords

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