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高分辨率开放式轴向扫描光片显微镜。

High-resolution open-top axially swept light sheet microscopy.

机构信息

Department of Mechanical Engineering, Pohang University of Science and Technology, 77 Cheongam-Ro, Nam-gu, Pohang, Gyeongbuk, 37673, Republic of Korea.

Department of Research and Development Center, Crayon Technologies, 19 Sanmaru-ro, Guri, Gyeonggi-do, 11901, Republic of Korea.

出版信息

BMC Biol. 2023 Nov 8;21(1):248. doi: 10.1186/s12915-023-01747-3.

Abstract

BACKGROUND

Open-top light-sheet microscopy (OT-LSM) is a specialized microscopic technique for the high-throughput cellular imaging of optically cleared, large-sized specimens, such as the brain. Despite the development of various OT-LSM techniques, achieving submicron resolution in all dimensions remains.

RESULTS

We developed a high-resolution open-top axially swept LSM (HR-OTAS-LSM) for high-throughput and high-resolution imaging in all dimensions. High axial and lateral resolutions were achieved by using an aberration-corrected axially swept excitation light sheet in the illumination arm and a high numerical aperture (NA) immersion objective lens in the imaging arm, respectively. The high-resolution, high-throughput visualization of neuronal networks in mouse brain and retina specimens validated the performance of HR-OTAS-LSM.

CONCLUSIONS

The proposed HR-OTAS-LSM method represents a significant advancement in the high-resolution mapping of cellular networks in biological systems such as the brain and retina.

摘要

背景

开顶式光片显微镜(OT-LSM)是一种专门的显微技术,用于对经过光学透明处理的大型标本(如大脑)进行高通量的细胞成像。尽管已经开发出了各种 OT-LSM 技术,但在所有维度上实现亚微米分辨率仍然具有挑战性。

结果

我们开发了一种用于高分辨率、高通量全维度成像的高分辨率开顶轴向扫描 LSM(HR-OTAS-LSM)。通过在照明臂中使用经过像差校正的轴向扫描激发光片和在成像臂中使用高数值孔径(NA)浸没物镜,实现了高轴向和高横向分辨率。通过对小鼠脑和视网膜标本中的神经元网络进行高分辨率、高通量可视化,验证了 HR-OTAS-LSM 的性能。

结论

所提出的 HR-OTAS-LSM 方法代表了在大脑和视网膜等生物系统中对细胞网络进行高分辨率映射的重大进展。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa91/10634022/77c56cd58346/12915_2023_1747_Fig1_HTML.jpg

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