Sparse deconvolution improves the resolution of live-cell super-resolution fluorescence microscopy

• Published in: Nature biotechnology Vol. 40; no. 4; pp. 606 - 617
• Main Authors: Zhao, Weisong, Zhao, Shiqun, Li, Liuju, Huang, Xiaoshuai, Xing, Shijia, Zhang, Yulin, Qiu, Guohua, Han, Zhenqian, Shang, Yingxu, Sun, De-en, Shan, Chunyan, Wu, Runlong, Gu, Lusheng, Zhang, Shuwen, Chen, Riwang, Xiao, Jian, Mo, Yanquan, Wang, Jianyong, Ji, Wei, Chen, Xing, Ding, Baoquan, Liu, Yanmei, Mao, Heng, Song, Bao-Liang, Tan, Jiubin, Liu, Jian, Li, Haoyu, Chen, Liangyi
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01-04-2022; Nature Publishing Group
• Subjects: 631/80; 639/624/1107/328/2238; Agriculture; Algorithms; Bioinformatics; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biomedicine; Biotechnology; Deconvolution; Fluorescence; Fluorescence microscopy; Imaging, Three-Dimensional - methods; Life Sciences; Microscopes; Microscopy; Microscopy, Fluorescence - methods; Mitochondria; Nuclear fusion; Nuclear pores; Nucleoporins; Photons; Pores; Spatial discrimination; Spatial resolution
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/s41587-021-01092-2

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.