西湖大学章永登等研究人员合作利用4Pi-SIM，在各向同性100纳米分辨率下阐明了亚细胞结构和动力学。相关论文于2024年12月23日在线发表在《自然—方法学》杂志上。

据介绍，3D-SIM提供了优异的光学切片功能，并且在所有维度上相较于宽场显微镜分辨率提高了一倍。然而，其轴向分辨率较低，导致该方向上的细节模糊并且整体图像失真。

研究人员介绍了4Pi-SIM，这是一种对I⁵S的重大改进，将三维结构照明显微镜（3D-SIM）与干涉显微镜结合，并通过照明和探测波前的干涉实现各向同性的光学分辨率。研究人员通过高保真度成像，不同细胞类型的亚细胞结构来评估4Pi-SIM的性能。

此外，研究人员通过进行数百个时间点的时间-体积成像，展示了其能力，达到了大约100纳米的三维分辨率。研究人员还展示了它同时进行双色成像的能力，并能够捕捉到紧密排列的细胞器之间的快速相互作用。

这些结果凸显了4Pi-SIM在阐明亚细胞结构，和揭示纳米尺度下动态行为方面的巨大潜力。

附：英文原文

Title: Elucidating subcellular architecture and dynamics at isotropic 100-nm resolution with 4Pi-SIM

Author: Ouyang, Zijing, Wang, Qian, Li, Xiaoyu, Dai, Qiuyang, Tang, Muyuan, Shao, Lin, Gou, Wen, Yu, Zijing, Chen, Yanqin, Zheng, Bei, Chen, Linlin, Ping, Conghui, Bi, Xiuli, Xiao, Bin, Yu, Xiaochun, Liu, Changliang, Chen, Liangyi, Fan, Junchao, Huang, Xiaoshuai, Zhang, Yongdeng

Issue&Volume: 2024-12-23

Abstract: Three-dimensional structured illumination microscopy (3D-SIM) provides excellent optical sectioning and doubles the resolution in all dimensions compared with wide-field microscopy. However, its much lower axial resolution results in blurred fine details in that direction and overall image distortion. Here we present 4Pi-SIM, a substantial revamp of I5S that synergizes 3D-SIM with interferometric microscopy to achieve isotropic optical resolution through interference in both the illumination and detection wavefronts. We evaluate the performance of 4Pi-SIM by imaging various subcellular structures across different cell types with high fidelity. Furthermore, we demonstrate its capability by conducting time-lapse volumetric imaging over hundreds of time points, achieving a 3D resolution of approximately 100nm. Additionally, we illustrate its ability to simultaneously image in two colors and capture the rapid interactions between closely positioned organelles in three dimensions. These results underscore the great potential of 4Pi-SIM for elucidating subcellular architecture and revealing dynamic behaviors at the nanoscale.

DOI: 10.1038/s41592-024-02515-z

Source:https://www.nature.com/articles/s41592-024-02515-z