A protocol for single-source dual-pulse stimulated emission depletion setup with Bessel modulation

Xinzhu Xu; Kun Zhao; Wei Ren; Zhaoyang Wu; Wentao Yu; Chendi Shao; Kebin Shi; Peng Xi

doi:10.1002/jemt.23922

A protocol for single-source dual-pulse stimulated emission depletion setup with Bessel modulation

Microsc Res Tech. 2022 Feb;85(2):813-823. doi: 10.1002/jemt.23922. Epub 2021 Sep 6.

Authors

Xinzhu Xu^{1

2}, Kun Zhao^{1

2}, Wei Ren¹, Zhaoyang Wu¹, Wentao Yu³, Chendi Shao³, Kebin Shi^{3

4}, Peng Xi^{1

5

6}

Affiliations

¹ Department of Biomedical Engineering, College of Future Technology, Peking University, Beijing, People's Republic of China.
² Wallace H Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia, USA.
³ State Key Laboratory for Mesoscopic Physics and Frontiers Science Center for Nano-optoelectronics, School of Physics, Peking University, Beijing, People's Republic of China.
⁴ Collaborative Innovation Center of Extreme Optics, Shanxi University, Taiyuan, People's Republic of China.
⁵ UTS-SUStech Joint Research Centre for Biomedical Materials & Devices, Department of Biomedical Engineering, College of Engineering, Southern University of Science and Technology, Shenzhen, Guangdong, People's Republic of China.
⁶ National Biomedical Imaging Center, Peking University, Beijing, People's Republic of China.

PMID: 34488243
DOI: 10.1002/jemt.23922

Abstract

STimulated Emission Depletion (STED) microscopy attains super-resolution in biological imaging beyond the diffraction limit. Here, we give a concise protocol to construct a dual-pulse STED setup with one super-continuum laser. Moreover, a flexible and dismountable Bessel modulation module is introduced for potential 2D-stack STED imaging. Experiments and notices are introduced in detail, with discussion on some important check-points for STED, such as detector saturation. Finally, the results validate the system working.

Keywords: Bessel modulation; STED microscopy; detector saturation.

MeSH terms

Lasers*
Microscopy, Fluorescence

Abstract

MeSH terms

Grants and funding