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在功能齐全的商用显微镜基座上实现用户友好型斜平面显微镜技术。

User-friendly oblique plane microscopy on a fully functional commercially available microscope base.

作者信息

Sirinakis George, Allgeyer Edward S, Nashchekin Dmitry, St Johnston Daniel

机构信息

The Gurdon Institute and the Department of Genetics University of Cambridge, Tennis Court Road, Cambridge, CB2 1QN, United Kingdom.

出版信息

Biomed Opt Express. 2024 Mar 14;15(4):2358-2376. doi: 10.1364/BOE.518856. eCollection 2024 Apr 1.

Abstract

In this work we present an oblique plane microscope designed to work seamlessly with a commercially available microscope base. To support all the functionality offered by the microscope base, where the position of the objective lens is not fixed, we adopted a two-mirror scanning geometry that can compensate for changes to the position of the objective lens during routine microscope operation. We showed that within a ± 1 mm displacement range of the 100X, 1.35 NA objective lens away from its designed position, the PSF size increased by <3% and <11% in the lateral and axial dimensions, respectively, while the error in magnification was <0.5% within volumes extending ± 10 µm about the focal plane. Compared to the more traditional scan-lens/galvo-mirror combination, the two-mirror scanning geometry offers higher light efficiency and a more compact footprint, which could be beneficial to all OPM designs regardless of the use of a commercial base or not.

摘要

在这项工作中,我们展示了一种设计用于与市售显微镜基座无缝配合工作的斜平面显微镜。为了支持显微镜基座所提供的所有功能,在物镜位置不固定的情况下,我们采用了双镜扫描几何结构,该结构能够补偿常规显微镜操作过程中物镜位置的变化。我们表明,在100X、1.35 NA物镜偏离其设计位置±1 mm的位移范围内,横向和轴向尺寸的点扩散函数(PSF)大小分别增加<3%和<11%,而在焦平面周围±10 µm范围内的体积中,放大误差<0.5%。与更传统的扫描透镜/振镜组合相比,双镜扫描几何结构具有更高的光效率和更紧凑的占地面积,这对所有斜平面显微镜(OPM)设计都可能有益,无论是否使用商业基座。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e0c2/11019673/296699043bd7/boe-15-4-2358-g001.jpg

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