1. Quantitative structured illumination microscopy via a physical model-based background filtering algorithm reveals actin dynamics.
- Author
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Mo, Yanquan, Wang, Kunhao, Li, Liuju, Xing, Shijia, Ye, Shouhua, Wen, Jiayuan, Duan, Xinxin, Luo, Ziying, Gou, Wen, Chen, Tongsheng, Zhang, Yu-Hui, Guo, Changliang, Fan, Junchao, and Chen, Liangyi
- Subjects
MICROSCOPY ,HIGH resolution imaging ,ACTIN ,CELL imaging ,LIGHTING - Abstract
Despite the prevalence of superresolution (SR) microscopy, quantitative live-cell SR imaging that maintains the completeness of delicate structures and the linearity of fluorescence signals remains an uncharted territory. Structured illumination microscopy (SIM) is the ideal tool for live-cell SR imaging. However, it suffers from an out-of-focus background that leads to reconstruction artifacts. Previous post hoc background suppression methods are prone to human bias, fail at densely labeled structures, and are nonlinear. Here, we propose a physical model-based Background Filtering method for living cell SR imaging combined with the 2D-SIM reconstruction procedure (BF-SIM). BF-SIM helps preserve intricate and weak structures down to sub-70 nm resolution while maintaining signal linearity, which allows for the discovery of dynamic actin structures that, to the best of our knowledge, have not been previously monitored. Quantitative live-cell superresolution imaging that maintains the linearity of fluorescence signals remains difficult. Here, the authors propose a physical model-based background filtering method for 2D-SIM, which allows for quantitative imaging and high signal completeness. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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