Single-molecule localization microscopy (SMLM) enables three-dimensional (3D) investigation of nanoscale structures in biological samples, offering unique insights into their organization. However, traditional 3D super-resolution microscopy using high numerical aperture (NA) objectives is limited by imaging depth of field (DOF), restricting their practical application to relatively thin biological samples. Here, we developed a unified solution for thick sample super-resolution imaging using a deformable mirror (DM) which served for fast remote focusing, optimized point spread function (PSF) engineering, and accurate aberration correction. By effectively correcting the system aberrations introduced during remote focusing and sample aberrations at different imaging depths, we achieved high-accuracy, large DOF imaging (∼8 μm$\sim 8\mathrm{\mu m}$) of the whole-cell organelles [i.e., nuclear pore complex (NPC), microtubules, and mitochondria] with a nearly uniform resolution of approximately 35 nm across the entire cellular volume.

中文翻译：

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基于可变形镜的远程聚焦的像差校正可实现高精度全细胞超分辨率成像

单分子定位显微镜 (SMLM) 能够对生物样品中的纳米级结构进行三维 (3D) 研究，提供对其组织的独特见解。然而，使用高数值孔径 (NA) 物镜的传统 3D 超分辨率显微镜受到成像景深 (DOF) 的限制，限制了其在相对较薄的生物样品上的实际应用。在这里，我们开发了一种使用可变形镜（DM）进行厚样品超分辨率成像的统一解决方案，可实现快速远程聚焦、优化的点扩散函数（PSF）工程和精确的像差校正。通过有效校正远程聚焦过程中引入的系统像差和不同成像深度的样品像差，我们实现了全细胞细胞器[即核孔复合体（NPC）、微管、 $\sim 8\mathrm{\mu m}$和线粒体]在整个细胞体积中具有大约35 nm的几乎一致的分辨率。