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.

中文翻译：

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一步 4× 和 12× 3D-ExM 可实现纳米级细胞结构的稳健超分辨率显微镜检查。


超分辨率显微镜已成为不同研究领域不可或缺的工具，以纳米级分辨率为生物结构提供了前所未有的见解。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项优势，包括同时标记多个目标生物分子，特异性高，样品制备更简单，使大多数研究人员都能使用。在这项研究中，我们介绍了两种优化的超分辨率成像方法：4 倍和 12 倍 3D 各向同性和保留扩增显微镜（4× 和 12× 3D-ExM）。3D-ExM 是一种简单的扩增显微镜技术，采用单步工艺，可为 2D 和 3D 细胞培养模型提供稳定且可重现的 3D 各向同性扩增。使用标准共聚焦显微镜，12× 3D-ExM 可实现 <30 nm 的横向分辨率，能够可视化纳米级结构，包括染色体、着丝粒、核孔复合物和 Epstein-Barr 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，包括细胞和染色质结构的研究。