Mirror-enhanced super-resolution microscopy

Xusan Yang; Hao Xie; Eric Alonas; Yujia Liu; Xuanze Chen; Philip J Santangelo; Qiushi Ren; Peng Xi; Dayong Jin

doi:10.1038/lsa.2016.134

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Mirror-enhanced super-resolution microscopy

Original Articles

- Mirror-enhanced super-resolution microscopy
- Mirror-enhanced super-resolution microscopy
- LSA 2016年5卷第3期
- Affiliations：
  
  1.Department of Biomedical Engineering, College of Engineering, Peking University, No. 5 Yiheyuan Road, Beijing 100871, China
  2.Wallace H Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USA
  3.Advanced Cytometry Labs, ARC Centre of Excellence for Nanoscale BioPhotonics (CNBP), Macquarie University, Sydney, NSW 2109, Australia
  4.Institute for Biomedical Materials and Devices (IBMD), Faculty of Science, University of Technology Sydney, Sydney, NSW 2007, Australia
- Author bio：
  
  P Xi, Email: xipeng@pku.edu.cn
- Funds：
- DOI：10.1038/lsa.2016.134
  中图分类号：
- 纸质出版日期：2016-09，
  
  网络出版日期：2016-06-17，
  
  收稿日期：2015-07-24，
  
  修回日期：2016-03-06，
  
  录用日期：2016-03-08
- Accepted：
Scan QR Code
Mirror-enhanced super-resolution microscopy[J]. LSA, 2016,5(3).

Yang, X. S. et al. Mirror-enhanced super-resolution microscopy. Light: Science & Applications, 5, e16134 (2016).
Mirror-enhanced super-resolution microscopy[J]. LSA, 2016,5(3). DOI： 10.1038/lsa.2016.134.

Yang, X. S. et al. Mirror-enhanced super-resolution microscopy. Light: Science & Applications, 5, e16134 (2016). DOI： 10.1038/lsa.2016.134.

摘要

Abstract

Axial excitation confinement beyond the diffraction limit is crucial to the development of next-generation

super-resolution microscopy. STimulated Emission Depletion (STED) nanoscopy offers lateral super-resolution using a donut-beam depletion

but its axial resolution is still over 500 nm. Total internal reflection fluorescence microscopy is widely used for single-molecule localization

but its ability to detect molecules is limited to within the evanescent field of ~100 nm from the cell attachment surface. We find here that the axial thickness of the point spread function (PSF) during confocal excitation can be easily improved to 110 nm by replacing the microscopy slide with a mirror. The interference of the local electromagnetic field confined the confocal PSF to a 110-nm spot axially

which enables axial super-resolution with all laser-scanning microscopes. Axial sectioning can be obtained with wavelength modulation or by controlling the spacer between the mirror and the specimen. With no additional complexity

the mirror-assisted excitation confinement enhanced the axial resolution six-fold and the lateral resolution two-fold for STED

which together achieved 19-nm resolution to resolve the inner rim of a nuclear pore complex and to discriminate the contents of 120 nm viral filaments. The ability to increase the lateral resolution and decrease the thickness of an axial section using mirror-enhanced STED without increasing the laser power is of great importance for imaging biological specimens

which cannot tolerate high laser power.

关键词

Keywords

confocalinterferencepoint spread functionsuper-resolution

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