Garbellotto Chiara, Taylor Jonathan M
University of Glasgow, University Avenue, Glasgow, G12 8QQ, UK.
Biomed Opt Express. 2018 Oct 12;9(11):5419-5436. doi: 10.1364/BOE.9.005419. eCollection 2018 Nov 1.
By integrating a phase-only Spatial Light Modulator (SLM) into the illumination arm of a cylindrical-lens-based Selective Plane Illumination Microscope (SPIM), we have created a versatile system able to deliver high quality images by operating in a wide variety of different imaging modalities. When placed in a Fourier plane, the SLM permits modulation of the microscope's light-sheet to implement imaging techniques such as structured illumination, tiling, pivoting, autofocusing and pencil beam scanning. Previous publications on dedicated microscope setups have shown how these techniques can deliver improved image quality by rejecting out-of-focus light (structured illumination and pencil beam scanning), reducing shadowing (light-sheet pivoting), and obtaining a more uniform illumination by moving the highest-resolution region of the light-sheet across the imaging Field of View (tiling). Our SLM-SPIM configuration is easy to build and use, and has been designed to allow all of these techniques to be employed on an easily reconfigurable optical setup, compatible with the OpenSPIM design. It offers the possibility to choose between three different light-sheets, in thickness and height, which can be selected according to the characteristics of the sample and the imaging technique to be applied. We demonstrate the flexibility and performance of the system with results obtained by applying a variety of different imaging techniques on samples of fluorescent beads, zebrafish embryos, and optically cleared whole mouse brain samples. Thus our approach allows easy implementation of advanced imaging techniques while retaining the simplicity of a cylindrical-lens-based light-sheet microscope.
通过将纯相位空间光调制器（SLM）集成到基于柱面透镜的选择性平面照明显微镜（SPIM）的照明臂中，我们创建了一个多功能系统，该系统能够通过多种不同的成像模式来提供高质量图像。当置于傅里叶平面时，SLM允许对显微镜的光片进行调制，以实现诸如结构照明、平铺、旋转、自动聚焦和笔形光束扫描等成像技术。先前关于专用显微镜设置的出版物已经展示了这些技术如何通过排除离焦光（结构照明和笔形光束扫描）、减少阴影（光片旋转）以及通过将光片的最高分辨率区域移动穿过成像视场（平铺）来获得更均匀的照明，从而提高图像质量。我们的SLM-SPIM配置易于构建和使用，并且设计为允许在易于重新配置的光学设置上采用所有这些技术，与OpenSPIM设计兼容。它提供了在三种不同厚度和高度的光片中进行选择的可能性，可以根据样品的特性和要应用的成像技术来选择。我们通过对荧光珠、斑马鱼胚胎和光学透明的全脑小鼠样本应用多种不同的成像技术所获得的结果，展示了该系统的灵活性和性能。因此，我们的方法允许轻松实现先进的成像技术，同时保留基于柱面透镜的光片显微镜的简单性。
