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| 基于艾里光束的开顶光片显微成像研究 |
| Research on microscopic imaging of open top light sheet based on Airy beam |
| 投稿时间：2024-04-02 修订日期：2024-04-17 |
| DOI： |
| 中文关键词: 开顶光片荧光显微镜 光束调制 Airy光束 轴向分辨率 |
| 英文关键词:open-top light sheet fluorescence microscopy beam modulation Airy beam axial resolution |
| 基金项目:国家重点研发计划项目(2021YFF1200600)、国家自然科学基金项目(批准号:62205242、91859101、81971744、U1932107、814717866、11804248)、天津市自然科学基金项目(21JCBJC00460)、天津市公共卫生科技重大专项(21ZXGWSY00070)、天津大学自主创新基金项目(2023XYY-0040)资助。 |
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| 中文摘要: |
| 光片荧光显微镜是一种新型光学成像方法，其照明路径和检测路径的正交特性，可以实现高时空分辨率、低光毒性和光漂白的三维成像。但是，目前的光片荧光显微镜大多采用传统的搭建方式，即：照明物镜被固定在与标本相平行的水平面上，然后放置检测物镜与其正交，但这种方式会约束样本尺寸及其横向移动，无法对厘米级的大尺寸样本进行多角度成像分析。这里，我们通过优化成像系统并结合光束调制手段构建了一套基于无衍射光束的开顶式光片成像系统。使用位于样本下方的三个物镜对样本进行光学层切并实现了高-低分辨率跨尺度成像。仿真结果以及标定实验证明了成像视场及分辨率提升的有效性。此外，我们还通过小鼠创伤性脑损伤模型的成像分析，展示了该成像系统在临床疾病研究与治疗中应用的巨大潜力。 |
| 英文摘要: |
| Light sheet fluorescence microscopy is a new optical imaging method. The orthogonal characteristics of illumination path and detection path can realize 3D imaging with high spatial and temporal resolution, low phototoxicity and light bleaching. However, most of the current light sheet fluorescence microscopes use the traditional construction method, that is, the illumination objective is fixed on the horizontal plane parallel to the specimen, and then the detection objective is placed orthogonal to it. However, this method will limit the sample size and its lateral movement, and it is impossible to perform multi-angle imaging analysis of the centimeter large sample. Here, we constructed an open-top light sheet imaging system based on a non-diffracting beam by optimizing the imaging system and combining the beam modulation. The sample was optically laminated and high-low resolution cross-scale imaging was achieved using three objective lenses positioned below the sample. Simulation results and calibration experiments demonstrate the effectiveness of improving the imaging field of view and resolution. In addition, we demonstrated the great potential of this imaging system for clinical disease research and treatment through imaging analysis of a mouse traumatic brain injury model. |
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