Visualizing intraorganellar ultrastructures, dynamics, and interactions with open-access background-free Lock-in-SIM

作者：Wenjie Liu, Meng Zhang, Wenbin Zhu, Shunyu Xie, Jinfeng Zhang, Shuhao Qian, Zhiyi Liu, Zheng Xiu, Qiuyuan Fang, Wei Yang, Yì Wáng, Dazhao Zhu, Jianjie Dong, Xü Liu, Youhua Chen, Cuifang Kuang, Yuhui Zhang, Lothar Schermelleh · 发表于：Nature Communications · 年份：2025 · DOI：10.1038/s41467-025-65805-w · 被引用次数：4 · 研究领域：Advanced Fluorescence Microscopy Techniques、Digital Holography and Microscopy、Cell Image Analysis Techniques

Structured illumination microscopy (SIM) is a powerful method for fast and gentle live-cell super-resolution imaging. However, its susceptibility to reconstruction artifacts from out-of-focus blur and background imposes substantial barriers to analyze the dynamics of densely packed volumetric intraorganellar ultrastructures that are typically in a size range of SIM's spatial resolution. To address this limitation, we have developed Lock-in-SIM, an open-access two-dimensional SIM framework that eliminates background and maximizes the recovery of sub-diffraction information with the highest possible frequency extraction. By leveraging the intrinsic modulation differences of volumetric sample structures, Lock-in-SIM enables efficient optical sectioning, extends imaging depth, and improves data fidelity and quantifiability. We demonstrate the superiority of Lock-in-SIM by visualizing various challenging intraorganellar ultrastructures in live cells. Our investigations uncover mechanisms of mitochondrial fission and endoplasmic reticulum-lysosome interactions and provide insights into the intricate yet highly regulated structural remodeling of organelles.