北京大学癌症研究所黄小帅团队实现了使用多平面结构照明显微镜进行快速、三维、活细胞超分辨率成像。2025年3月3日出版的《自然—光子学》杂志发表了这项成果。

三维结构照明显微镜（3D-SIM）使所有维度的空间分辨率加倍，并广泛应用于细胞成像。然而，其时间分辨率受到使用压电台进行成像的样品顺序逐平面移动的需要的限制，这通常会将每个体积的采集时间增加到几秒钟。

为了解决这一局限性，研究组开发了3D多平面SIM（3D-MP-SIM），它可以同时检测多平面图像，并使用协同进化的重建算法进行重建。与传统的3D-SIM成像相比，3D-MP-SIM的体积超分辨率成像的时间分辨率提高了大约八倍，横向和轴向空间分辨率分别约为120和300 nm。快速采集大大减少了活细胞中动态结构（如晚期内体）成像过程中的运动伪影。

此外，研究组通过以高达每秒11个体积的速率对内质网进行高速延时体积成像，展示了3D-MP-SIM的能力。还通过观察3D空间中细胞内和细胞间细胞器之间快速而密切的相互作用，展示了双色成像的可行性。这些结果突出了3D-MP-SIM在亚细胞水平和三维解释动态行为和相互作用方面的潜力。

附：英文原文

Title: Fast, three-dimensional, live-cell super-resolution imaging with multiplane structured illumination microscopy

Author: Chen, Qian, Gou, Wen, Lu, Wenqing, Li, Jie, Wei, Yuhong, Li, Haoyu, Wang, Chengyu, You, Wei, Li, Zhengqian, Dong, Dashan, Bi, Xiuli, Xiao, Bin, Chen, Liangyi, Shi, Kebin, Fan, Junchao, Huang, Xiaoshuai

Issue&Volume: 2025-03-03

Abstract: Three-dimensional structured illumination microscopy (3D-SIM) doubles the spatial resolution along all dimensions and is used widely in cellular imaging. However, its temporal resolution is constrained by the need for sequential plane-by-plane movement of the sample using a piezo stage for imaging, which often increases the acquisition time to several seconds per volume. To address this limitation, we develop 3D multiplane SIM (3D-MP-SIM), which simultaneously detects multiplane images and reconstructs them using synergistically evolved reconstruction algorithms. Compared with conventional 3D-SIM imaging, 3D-MP-SIM achieves an approximately eightfold increase in the temporal resolution of volumetric super-resolution imaging, with lateral and axial spatial resolutions of about 120 and 300nm, respectively. The rapid acquisition substantially reduces motion artefacts during the imaging of dynamic structures, such as late endosomes, in live cells. Moreover, we demonstrate the capabilities of 3D-MP-SIM via high-speed time-lapse volumetric imaging of the endoplasmic reticulum at rates of up to 11 volumes per second. We also show the feasibility of dual-colour imaging by observing rapid and close interactions among intra- and intercellular organelles in 3D space. These results highlight the potential of 3D-MP-SIM for explaining dynamic behaviours and interactions at the subcellular level and in three dimensions.

DOI: 10.1038/s41566-025-01638-9

Source: https://www.nature.com/articles/s41566-025-01638-9