Li Guofang, Ge Enxiang, Zheng Hua, Lin Weiying
Institute of Optical Materials and Chemical Biology, Guangxi Key Laboratory of Electrochemical Energy Materials, School of Chemistry and Chemical Engineering, Guangxi University Nanning Guangxi 530004 China
Chem Sci. 2025 Sep 8. doi: 10.1039/d5sc04640h.
As a cutting-edge super-resolution imaging technique, structured illumination microscopy (SIM) has been widely used in cell biology research, especially in the analysis of subcellular organelles and monitoring of their dynamic processes. Through multiple illumination and reconstruction processes, SIM breaks through the resolution limitations of traditional microscopes and can observe the fine structures within cells in real time with nanoscale resolution. This provides strong technical support for in-depth analyses of molecular mechanisms, organelle functions, signaling networks, and metabolic regulatory pathways within cells. In recent years, super-resolution imaging technology, as a novel imaging technique, has made significant research progress in subcellular fine structure imaging by combining fluorescent probes. However, there is an urgent need for a more comprehensive review of the use of fluorescent probes in various organelles (such as mitochondria, lysosomes, lipid droplets, cell membranes, and the endoplasmic reticulum) by SIM imaging technology. Therefore, this review provides a comprehensive overview of the latest research progress in observing different organelles using small-molecule fluorescent probes by SIM imaging. At the same time, we also discussed the challenges and prospects of current SIM technology in dynamic process observation, including further improvement of resolution, photobleaching issues in long-term observations, and innovation in multimodal imaging. Overall, this review fills the research gap in this important field by providing a comprehensive, multi-organelle-targeted fluorescent probe perspective, combined with SIM imaging technology, particularly providing readers with new ideas and insights at multiple levels, such as technology, design, application, and disease research.
作为一种前沿的超分辨率成像技术，结构光照明显微镜（SIM）已广泛应用于细胞生物学研究，尤其是在亚细胞器分析及其动态过程监测方面。通过多次照明和重建过程，SIM突破了传统显微镜的分辨率限制，能够以纳米级分辨率实时观察细胞内的精细结构。这为深入分析细胞内的分子机制、细胞器功能、信号网络和代谢调控途径提供了有力的技术支持。近年来，超分辨率成像技术作为一种新型成像技术，通过结合荧光探针在亚细胞精细结构成像方面取得了显著的研究进展。然而，迫切需要对SIM成像技术在各种细胞器（如线粒体、溶酶体、脂滴、细胞膜和内质网）中使用荧光探针的情况进行更全面的综述。因此，本综述全面概述了利用SIM成像技术使用小分子荧光探针观察不同细胞器的最新研究进展。同时，我们还讨论了当前SIM技术在动态过程观察中的挑战和前景，包括分辨率的进一步提高、长期观察中的光漂白问题以及多模态成像的创新。总体而言，本综述通过提供一个全面的、针对多细胞器的荧光探针视角，并结合SIM成像技术，填补了这一重要领域的研究空白，尤其为读者在技术、设计、应用和疾病研究等多个层面提供了新的思路和见解。
