Molecular-scale axial localization by repetitive optical selective exposure

Lusheng Gu^#^{1

2

3

4

5}, Yuanyuan Li^#^{1

2

5}, Shuwen Zhang^#^{1

2

4

6}, Maoge Zhou^{1

2

4}, Yanhong Xue^{1

2

4}, Weixing Li^{1

2

3

5}, Tao Xu^{7

8

9

10

11}, Wei Ji^{12

13

14

15

16}

Affiliations

¹ National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
² Physical Science Laboratory, Huairou National Comprehensive Science Center, Beijing, China.
³ Bioland Laboratory (Guangzhou Regenerative Medicine and Health Guangdong Laboratory), Guangzhou, Guangdong, China.
⁴ College of Life Science, University of Chinese Academy of Sciences, Beijing, China.
⁵ Core Facility for Protein Research, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
⁶ School of Future Technology, University of Chinese Academy of Sciences, Beijing, China.
⁷ National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China. xutao@ibp.ac.cn.
⁸ Physical Science Laboratory, Huairou National Comprehensive Science Center, Beijing, China. xutao@ibp.ac.cn.
⁹ Bioland Laboratory (Guangzhou Regenerative Medicine and Health Guangdong Laboratory), Guangzhou, Guangdong, China. xutao@ibp.ac.cn.
¹⁰ College of Life Science, University of Chinese Academy of Sciences, Beijing, China. xutao@ibp.ac.cn.
¹¹ School of Future Technology, University of Chinese Academy of Sciences, Beijing, China. xutao@ibp.ac.cn.
¹² National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China. jiwei@ibp.ac.cn.
¹³ Physical Science Laboratory, Huairou National Comprehensive Science Center, Beijing, China. jiwei@ibp.ac.cn.
¹⁴ Bioland Laboratory (Guangzhou Regenerative Medicine and Health Guangdong Laboratory), Guangzhou, Guangdong, China. jiwei@ibp.ac.cn.
¹⁵ College of Life Science, University of Chinese Academy of Sciences, Beijing, China. jiwei@ibp.ac.cn.
¹⁶ Core Facility for Protein Research, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China. jiwei@ibp.ac.cn.

^# Contributed equally.

PMID: 33795876
DOI: 10.1038/s41592-021-01099-2

Molecular-scale axial localization by repetitive optical selective exposure

Lusheng Gu et al. Nat Methods. 2021 Apr.

. 2021 Apr;18(4):369-373.

doi: 10.1038/s41592-021-01099-2. Epub 2021 Apr 1.

Authors

Affiliations

¹ National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
² Physical Science Laboratory, Huairou National Comprehensive Science Center, Beijing, China.
³ Bioland Laboratory (Guangzhou Regenerative Medicine and Health Guangdong Laboratory), Guangzhou, Guangdong, China.
⁴ College of Life Science, University of Chinese Academy of Sciences, Beijing, China.
⁵ Core Facility for Protein Research, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
⁶ School of Future Technology, University of Chinese Academy of Sciences, Beijing, China.
⁷ National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China. xutao@ibp.ac.cn.
⁸ Physical Science Laboratory, Huairou National Comprehensive Science Center, Beijing, China. xutao@ibp.ac.cn.
⁹ Bioland Laboratory (Guangzhou Regenerative Medicine and Health Guangdong Laboratory), Guangzhou, Guangdong, China. xutao@ibp.ac.cn.
¹⁰ College of Life Science, University of Chinese Academy of Sciences, Beijing, China. xutao@ibp.ac.cn.
¹¹ School of Future Technology, University of Chinese Academy of Sciences, Beijing, China. xutao@ibp.ac.cn.
¹² National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China. jiwei@ibp.ac.cn.
¹³ Physical Science Laboratory, Huairou National Comprehensive Science Center, Beijing, China. jiwei@ibp.ac.cn.
¹⁴ Bioland Laboratory (Guangzhou Regenerative Medicine and Health Guangdong Laboratory), Guangzhou, Guangdong, China. jiwei@ibp.ac.cn.
¹⁵ College of Life Science, University of Chinese Academy of Sciences, Beijing, China. jiwei@ibp.ac.cn.
¹⁶ Core Facility for Protein Research, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China. jiwei@ibp.ac.cn.

^# Contributed equally.

PMID: 33795876
DOI: 10.1038/s41592-021-01099-2

Abstract

We introduce an axial localization with repetitive optical selective exposure (ROSE-Z) method for super-resolution imaging. By using an asymmetric optical scheme to generate interference fringes, a <2 nm axial localization precision was achieved with only ~3,000 photons, which is an approximately sixfold improvement compared to previous astigmatism methods. Nanoscale three-dimensional and two-color imaging was demonstrated, illustrating how this method achieves superior performance and facilitates the investigation of cellular nanostructures.

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References

1. Betzig, E. et al. Imaging Intracellular fluorescent proteins at nanometer resolution. Science 313, 1642–1645 (2006). - DOI
1. Hess, S. T., Girirajan, T. P. K. & Mason, M. D. Ultra-high resolution imaging by fluorescence photoactivation localization microscopy. Biophysical J. 91, 4258–4272 (2006). - DOI
1. Rust, M. J., Bates, M. & Zhuang, X. Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat. Methods 3, 793 (2006). - DOI
1. Huang, B., Wang, W., Bates, M. & Zhuang, X. Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy. Science 319, 810–813 (2008). - DOI
1. Juette, M. F. et al. Three-dimensional sub-100 nm resolution fluorescence microscopy of thick samples. Nat. Methods 5, 527–529 (2008). - DOI

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Molecular-scale axial localization by repetitive optical selective exposure

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Molecular-scale axial localization by repetitive optical selective exposure

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