Wen Gang, Leen Volker, Rohand Taoufik, Sauer Markus, Hofkens Johan
Department of Chemistry, KU Leuven, Leuven 3001, Belgium.
Chrometra Scientific, Kortenaken 3470, Belgium.
Chem Rev. 2023 Mar 22;123(6):3299-3323. doi: 10.1021/acs.chemrev.2c00711. Epub 2023 Mar 7.
Expansion microscopy (ExM) is a newly developed super-resolution technique, allowing visualization of biological targets at nanoscale resolution on conventional fluorescence microscopes. Since its introduction in 2015, many efforts have been dedicated to broaden its application range or increase the resolution that can be achieved. As a consequence, recent years have witnessed remarkable advances in ExM. This review summarizes recent progress in ExM, with the focus on the chemical aspects of the method, from chemistries for biomolecule grafting to polymer synthesis and the impact on biological analysis. The combination of ExM with other microscopy techniques, in search of additional resolution improvement, is also discussed. In addition, we compare pre- and postexpansion labeling strategies and discuss the impact of fixation methods on ultrastructure preservation. We conclude this review with a perspective on existing challenges and future directions. We believe that this review will provide a comprehensive understanding of ExM and facilitate its usage and further development.
扩展显微镜技术（ExM）是一种新开发的超分辨率技术，可在传统荧光显微镜上以纳米级分辨率观察生物靶点。自2015年问世以来，人们付出了诸多努力来拓宽其应用范围或提高可实现的分辨率。因此，近年来扩展显微镜技术取得了显著进展。本综述总结了扩展显微镜技术的最新进展，重点关注该方法的化学方面，从生物分子接枝化学到聚合物合成及其对生物分析的影响。还讨论了扩展显微镜技术与其他显微镜技术的结合，以寻求进一步提高分辨率。此外，我们比较了膨胀前和膨胀后的标记策略，并讨论了固定方法对超微结构保存的影响。我们以对现有挑战和未来方向的展望结束本综述。我们相信，本综述将提供对扩展显微镜技术的全面理解，并促进其使用和进一步发展。
