近日，北京大学席鹏团队报道了用共焦旋转圆盘图像扫描显微镜实现高保真组织超分辨率成像。2025年8月4日，《光：科学与应用》杂志发表了这一成果。

超分辨率成像彻底改变了他们在亚细胞尺度上可视化生物结构的能力。然而，深层组织超分辨率成像仍然受到背景干扰的限制，导致深度穿透有限，成像保真度降低。

为了克服这些挑战，研究组提出了一种新的成像系统，共焦旋转磁盘图像扫描显微镜（C2SD-ISM）。它集成了一个旋转盘（SD）共聚焦显微镜，物理上消除了失焦信号，形成了第一个共聚焦水平。采用数字微镜器件（DMD）进行稀疏多焦照明，结合动态针孔阵列像素重分配（DPA-PR）算法进行ISM超分辨率重建，形成第二共焦层。

双共焦结构提高了系统分辨率，同时有效地减轻了散射背景干扰。与计算出的离焦信号去除相比，随着穿透深度的增加，SD保持了原始强度分布，成像深度可达180 μm。此外，DPA-PR算法有效地纠正了Stokes偏移、光学像差和其他非理想条件，实现了144轴向分辨率为351原始共焦图像和重建图像之间的线性相关性高达92%，从而实现高保真的超分辨率成像。

此外，该系统通过DMD的可编程照明允许与结构化照明显微镜模式无缝实现，提供出色的可扩展性和易于主题。总之，这些功能使C2SD-ISM系统成为一种多功能工具，为现代生物成像需求推进细胞成像和组织尺度探索。

附：英文原文

Title: High-fidelity tissue super-resolution imaging achieved with confocal2 spinning-disk image scanning microscopy

Author: Liang, Qianxi, Ren, Wei, Jin, Boya, Qiao, Liang, Ge, Xichuan, Fu, Yunzhe, Lv, Xiaoqi, Li, Meiqi, Xi, Peng

Issue&Volume: 2025-08-04

Abstract: Super-resolution imaging has revolutionized our ability to visualize biological structures at subcellular scales. However, deep-tissue super-resolution imaging remains constrained by background interference, which leads to limited depth penetration and compromised imaging fidelity. To overcome these challenges, we propose a novel imaging system, confocal2 spinning-disk image scanning microscopy (C2SD-ISM). It integrates a spinning-disk (SD) confocal microscope, which physically eliminates out-of-focus signals, forming the first confocal level. A digital micromirror device (DMD) is employed for sparse multifocal illumination, combined with a dynamic pinhole array pixel reassignment (DPA-PR) algorithm for ISM super-resolution reconstruction, forming the second confocal level. The dual confocal configuration enhances system resolution, while effectively mitigating scattering background interference. Compared to computational out-of-focus signal removal, SD preserves the original intensity distribution as the penetration depth increases, achieving an imaging depth of up to 180μm. Additionally, the DPA-PR algorithm effectively corrects Stokes shifts, optical aberrations, and other non-ideal conditions, achieving a lateral resolution of 144nm and an axial resolution of 351nm, and a linear correlation of up to 92% between the original confocal and the reconstructed image, thereby enabling high-fidelity super-resolution imaging. Moreover, the system’s programmable illumination via the DMD allows for seamless realization with structured illumination microscopy modality, offering excellent scalability and ease of use. Altogether, these capabilities make the C2SD-ISM system a versatile tool, advancing cellular imaging and tissue-scale exploration for modern bioimaging needs.

DOI: 10.1038/s41377-025-01930-x

Source: https://www.nature.com/articles/s41377-025-01930-x