Kumar Manish, Kozorovitskiy Yevgenia
Department of Neurobiology, Northwestern University, Evanston, Illinois 60208, USA.
Biomed Opt Express. 2020 May 28;11(6):3346-3359. doi: 10.1364/BOE.389654. eCollection 2020 Jun 1.
Oblique plane microscopy (OPM) is a single objective light-sheet microscopy that performs three dimensional (3D) imaging by axial scan of the generated light-sheet. Recently, multiple techniques for lateral scanning of the generated light-sheet in OPM have emerged. However, their suitability for geometrically distortion free 3D imaging, which essentially requires a constant tilt light-sheet scan, has not been evaluated. In this work, we use a geometrical optics approach and derive analytical relationship for the amount of tilt variance in planar mirror based scanned oblique plane illumination (SOPi) arrangement. We experimentally validate the derived relationship and use it to arrive at an optimized scanner geometry and to understand its associated limitations. We discuss the effects of scanning on optical aberrations and 3D field of view in optimized, tilt invariant, lateral scanning OPM systems. We also provide experimental strategies enabling precise scanner alignment for tilt invariance, as well as an open source platform for rapid design of new oblique light-sheet microscopes.
斜平面显微镜（OPM）是一种单物镜光片显微镜，通过对生成的光片进行轴向扫描来执行三维（3D）成像。最近，在OPM中出现了多种用于横向扫描生成光片的技术。然而，它们对于无几何失真的3D成像的适用性尚未得到评估，而这种成像本质上需要恒定倾斜的光片扫描。在这项工作中，我们使用几何光学方法，推导了基于平面镜的扫描斜平面照明（SOPi）装置中倾斜变化量的解析关系。我们通过实验验证了推导的关系，并利用它得出优化的扫描仪几何结构并了解其相关限制。我们讨论了扫描对优化的、倾斜不变的横向扫描OPM系统中的光学像差和3D视野的影响。我们还提供了实现倾斜不变性的精确扫描仪对准的实验策略，以及用于快速设计新型斜光片显微镜的开源平台。
