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。

IF 23.4 Q1 OPTICS
Qianxi Liang,Wei Ren,Boya Jin,Liang Qiao,Xichuan Ge,Yunzhe Fu,Xiaoqi Lv,Meiqi Li,Peng Xi
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引用次数: 0

摘要

超分辨率成像彻底改变了我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,我们提出了一种新的成像系统,共聚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级。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。与计算出的失焦信号去除相比,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算法有效地校正了Stokes位移、光学像差和其他非理想条件,实现了144 nm的横向分辨率和351nm的轴向分辨率,原始共焦图像和重建图像之间的线性相关性高达92%,从而实现了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用性。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。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
High-fidelity tissue super-resolution imaging achieved with confocal2 spinning-disk image scanning microscopy.
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 180 μm. Additionally, the DPA-PR algorithm effectively corrects Stokes shifts, optical aberrations, and other non-ideal conditions, achieving a lateral resolution of 144 nm and an axial resolution of 351 nm, 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.
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来源期刊
Light-Science & Applications
Light-Science & Applications 数理科学, 物理学I, 光学, 凝聚态物性 II :电子结构、电学、磁学和光学性质, 无机非金属材料, 无机非金属类光电信息与功能材料, 工程与材料, 信息科学, 光学和光电子学, 光学和光电子材料, 非线性光学与量子光学
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发文量
803
审稿时长
2.1 months
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