Multi-purpose SLM-light-sheet microscope.
Multi-purpose SLM-light-sheet microscope.
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DOI：
10.1364/boe.9.005419
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发表时间：
2018-11-01
影响因子：
3.4
通讯作者：
Taylor JM
中科院分区：
文献类型：
--
作者：
Garbellotto C;Taylor JM
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，该系统能够通过在各种不同的成像模式下操作来提供高质量的图像。当放置在傅立叶平面中时，SLM允许调制显微镜的光片以实现成像技术，例如结构化照明、平铺、枢转、自动聚焦和笔形光束扫描。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拒绝失焦光（结构化照明和笔形光束扫描）、减少阴影（光片枢转）以及通过将光片的最高分辨率区域移动到成像视场（平铺）来获得更均匀的照明来提供改进的图像质量。我们的SLM-SPIM配置易于构建和使用，并且旨在允许所有这些技术在易于重新配置的光学设置上使用，与OpenSPIM设计兼容。它提供了在三种不同的光片之间进行选择的可能性，厚度和高度可以根据样品的特性和要应用的成像技术进行选择。我们展示了系统的灵活性和性能，通过应用各种不同的成像技术对荧光珠，斑马鱼胚胎和光学透明的整个小鼠大脑样品的样品所获得的结果。因此，我们的方法可以很容易地实现先进的成像技术，同时保持简单的圆柱透镜为基础的光片显微镜。
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.