Chinese Optics Letters, 2024, 22 (3): 031701, Published Online: Mar. 14, 2024  

Stimulated emission–depletion-based point-scanning structured illumination microscopy

Author Affiliations
1 Guangdong Key Laboratory for Biomedical Measurements and Ultrasound Imaging, National-Regional Key Technology Engineering Laboratory for Medical Ultrasound, School of Biomedical Engineering, Shenzhen University Medical School, Shenzhen 518060, China
2 Key Laboratory of Opto-electronic Information Science and Technology of Jiangxi Province, Nanchang Hangkong University, Nanchang 330063, China
3 College of Physics and Optoelectronics Engineering, Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, Shenzhen University, Shenzhen 518060, China
4 Department of Bioengineering and COMSET, Clemson University, Clemson SC 29634, US
Abstract
Wide-field linear structured illumination microscopy (LSIM) extends resolution beyond the diffraction limit by moving unresolvable high-frequency information into the passband of the microscopy in the form of moiré fringes. However, due to the diffraction limit, the spatial frequency of the structured illumination pattern cannot be larger than the microscopy cutoff frequency, which results in a twofold resolution improvement over wide-field microscopes. This Letter presents a novel approach in point-scanning LSIM, aimed at achieving higher-resolution improvement by combining stimulated emission depletion (STED) with point-scanning structured illumination microscopy (psSIM) (STED-psSIM). The according structured illumination pattern whose frequency exceeds the microscopy cutoff frequency is produced by scanning the focus of the sinusoidally modulated excitation beam of STED microscopy. The experimental results showed a 1.58-fold resolution improvement over conventional STED microscopy with the same depletion laser power.

Lei Wang, Meiting Wang, Luwei Wang, Xiaomin Zheng, Jiajie Chen, Wenshuai Wu, Wei Yan, Bin Yu, Junle Qu, Bruce Zhi Gao, Yonghong Shao. Stimulated emission–depletion-based point-scanning structured illumination microscopy[J]. Chinese Optics Letters, 2024, 22(3): 031701.

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