Advancements and perspectives on organelle-targeted fluorescent probes for super-resolution SIM imaging.

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.