Content area
Abstract
A main determinant of the spatial resolution of live-cell super-resolution (SR) microscopes is the maximum photon flux that can be collected. To further increase the effective resolution for a given photon flux, we take advantage of a priori knowledge about the sparsity and continuity of biological structures to develop a deconvolution algorithm that increases the resolution of SR microscopes nearly twofold. Our method, sparse structured illumination microscopy (Sparse-SIM), achieves ~60-nm resolution at a frame rate of up to 564 Hz, allowing it to resolve intricate structures, including small vesicular fusion pores, ring-shaped nuclear pores formed by nucleoporins and relative movements of inner and outer mitochondrial membranes in live cells. Sparse deconvolution can also be used to increase the three-dimensional resolution of spinning-disc confocal-based SIM, even at low signal-to-noise ratios, which allows four-color, three-dimensional live-cell SR imaging at ~90-nm resolution. Overall, sparse deconvolution will be useful to increase the spatiotemporal resolution of live-cell fluorescence microscopy.
The resolution of fluorescence microscopy is increased by incorporating prior information into deconvolution algorithms.
Details
; Zhao Shiqun 2 ; Li Liuju 2 ; Huang Xiaoshuai 3 ; Xing Shijia 2 ; Zhang, Yulin 2
; Qiu Guohua 1 ; Han Zhenqian 1 ; Shang Yingxu 4 ; De-en, Sun 5
; Shan Chunyan 6
; Wu Runlong 2 ; Gu Lusheng 7 ; Zhang, Shuwen 7 ; Chen Riwang 8 ; Xiao, Jian 9 ; Mo Yanquan 2 ; Wang, Jianyong 8 ; Ji, Wei 7 ; Chen, Xing 5
; Ding Baoquan 4
; Liu, Yanmei 10 ; Mao Heng 11 ; Bao-Liang, Song 9
; Tan Jiubin 12 ; Liu, Jian 13 ; Li, Haoyu 14
; Chen, Liangyi 15
1 Harbin Institute of Technology, Advanced Microscopy and Instrumentation Research Center, School of Instrumentation Science and Engineering, Harbin, China (GRID:grid.19373.3f) (ISNI:0000 0001 0193 3564)
2 Peking University, State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, National Biomedical Imaging Center, School of Future Technology, Beijing, China (GRID:grid.11135.37) (ISNI:0000 0001 2256 9319)
3 Peking University, Biomedical Engineering Department, Beijing, China (GRID:grid.11135.37) (ISNI:0000 0001 2256 9319)
4 National Center for Nanoscience and Technology, CAS Key Laboratory of Nanosystem and Hierarchical Fabrication, CAS Center for Excellence in Nanoscience, Beijing, China (GRID:grid.419265.d) (ISNI:0000 0004 1806 6075)
5 Peking University, College of Chemistry and Molecular Engineering, Beijing, China (GRID:grid.11135.37) (ISNI:0000 0001 2256 9319)
6 Peking University, College of Life Sciences, Beijing, China (GRID:grid.11135.37) (ISNI:0000 0001 2256 9319)
7 Chinese Academy of Sciences, National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Beijing, China (GRID:grid.9227.e) (ISNI:0000000119573309)
8 Peking University, School of Software and Microelectronics, Beijing, China (GRID:grid.11135.37) (ISNI:0000 0001 2256 9319)
9 Wuhan University, Hubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan, China (GRID:grid.49470.3e) (ISNI:0000 0001 2331 6153)
10 Peking University, State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, National Biomedical Imaging Center, School of Future Technology, Beijing, China (GRID:grid.11135.37) (ISNI:0000 0001 2256 9319); South China Normal University, Institute for Brain Research and Rehabilitation (IBRR), Guangdong Key Laboratory of Mental Health and Cognitive Science, Guangzhou, China (GRID:grid.263785.d) (ISNI:0000 0004 0368 7397)
11 Peking University, School of Mathematical Sciences, Beijing, China (GRID:grid.11135.37) (ISNI:0000 0001 2256 9319)
12 Harbin Institute of Technology, Center of Ultra-Precision Optoelectronic Instrument Engineering, Harbin, China (GRID:grid.19373.3f) (ISNI:0000 0001 0193 3564); Harbin Institute of Technology, Key Laboratory of Ultra-Precision Intelligent Instrumentation of Ministry of Industry and Information Technology, Harbin, China (GRID:grid.19373.3f) (ISNI:0000 0001 0193 3564)
13 Harbin Institute of Technology, Advanced Microscopy and Instrumentation Research Center, School of Instrumentation Science and Engineering, Harbin, China (GRID:grid.19373.3f) (ISNI:0000 0001 0193 3564); Harbin Institute of Technology, Key Laboratory of Micro-Systems and Micro-Structures Manufacturing of Ministry of Education, Harbin, China (GRID:grid.19373.3f) (ISNI:0000 0001 0193 3564)
14 Harbin Institute of Technology, Advanced Microscopy and Instrumentation Research Center, School of Instrumentation Science and Engineering, Harbin, China (GRID:grid.19373.3f) (ISNI:0000 0001 0193 3564); Harbin Institute of Technology, Key Laboratory of Ultra-Precision Intelligent Instrumentation of Ministry of Industry and Information Technology, Harbin, China (GRID:grid.19373.3f) (ISNI:0000 0001 0193 3564); Harbin Institute of Technology, Laboratory for Space Environment and Physical Sciences, Harbin, China (GRID:grid.19373.3f) (ISNI:0000 0001 0193 3564)
15 Peking University, State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, National Biomedical Imaging Center, School of Future Technology, Beijing, China (GRID:grid.11135.37) (ISNI:0000 0001 2256 9319); PKU-IDG/McGovern Institute for Brain Research, Beijing, China (GRID:grid.11135.37) (ISNI:0000 0001 2256 9319); Beijing Academy of Artificial Intelligence, Beijing, China (GRID:grid.511045.4)