Norman Roshan X, Chen Yu-Chia, Recchia Emma E, Loi Jonathan, Rosemarie Quincy, Lesko Sydney L, Patel Smit, Sherer Nathan, Takaku Motoki, Burkard Mark E, Suzuki Aussie
Biophysics Graduate Program, University of Wisconsin-Madison , Madison, WI, USA.
Department of Oncology, McArdle Laboratory for Cancer Research, University of Wisconsin-Madison, Madison, WI, USA.
J Cell Biol. 2025 Feb 3;224(2). doi: 10.1083/jcb.202407116. Epub 2024 Dec 3.
Super-resolution microscopy has become an indispensable tool across diverse research fields, offering unprecedented insights into biological architectures with nanometer scale resolution. Compared with traditional nanometer-scale imaging methods such as electron microscopy, super-resolution microscopy offers several advantages, including the simultaneous labeling of multiple target biomolecules with high specificity and simpler sample preparation, making it accessible to most researchers. In this study, we introduce two optimized methods of super-resolution imaging: 4-fold and 12-fold 3D-isotropic and preserved Expansion Microscopy (4× and 12× 3D-ExM). 3D-ExM is a straightforward expansion microscopy technique featuring a single-step process, providing robust and reproducible 3D isotropic expansion for both 2D and 3D cell culture models. With standard confocal microscopy, 12× 3D-ExM achieves a lateral resolution of <30 nm, enabling the visualization of nanoscale structures, including chromosomes, kinetochores, nuclear pore complexes, and Epstein-Barr virus particles. These results demonstrate that 3D-ExM provides cost-effective and user-friendly super-resolution microscopy, making it highly suitable for a wide range of cell biology research, including studies on cellular and chromatin architectures.
超分辨率显微镜已成为各个研究领域不可或缺的工具，能够以纳米级分辨率为生物结构提供前所未有的见解。与电子显微镜等传统纳米级成像方法相比，超分辨率显微镜具有多个优势，包括能够以高特异性同时标记多个目标生物分子以及更简单的样品制备，这使得大多数研究人员都能够使用。在本研究中，我们介绍了两种优化的超分辨率成像方法：4倍和12倍的三维各向同性和保留型扩展显微镜（4×和12× 3D-ExM）。3D-ExM是一种简单的扩展显微镜技术，具有单步流程，可为二维和三维细胞培养模型提供强大且可重复的三维各向同性扩展。使用标准共聚焦显微镜，12× 3D-ExM可实现小于30纳米的横向分辨率，能够可视化纳米级结构，包括染色体、动粒、核孔复合体和爱泼斯坦-巴尔病毒颗粒。这些结果表明，3D-ExM提供了经济高效且用户友好的超分辨率显微镜，使其非常适合广泛的细胞生物学研究，包括对细胞结构和染色质结构的研究。
