Institute for Biology, Division of Developmental Genetics, University of Kassel, Heinrich-Plett Str. 40, 34132, Kassel, Germany.
Department of Physics, Institute for Advanced Studies in Basic Sciences (IASBS), Zanjan, 45137-66731, Iran.
Sci Rep. 2018 Jul 3;8(1):10067. doi: 10.1038/s41598-018-28389-8.
Selective plane illumination microscopy (SPIM) represents a preferred method in dynamic tissue imaging, because it combines high spatiotemporal resolution with low phototoxicity. The OpenSPIM system was developed to provide an accessible and flexible microscope set-up for non-specialist users. Here, we report Structured SPIM (SSPIM), which offers an open-source, user-friendly and compact toolbox for beam shaping to be applied within the OpenSPIM platform. SSPIM is able to generate digital patterns for a wide range of illumination beams including static and spherical Gaussian beams, Bessel beams and Airy beams by controlling the pattern of a Spatial Light Modulator (SLM). In addition, SSPIM can produce patterns for structured illumination including incoherent and coherent array beams and tiling for all types of the supported beams. We describe the workflow of the toolbox and demonstrate its application by comparing experimental data with simulation results for a wide range of illumination beams. Finally, the capability of SSPIM is investigated by 3D imaging of Drosophila embryos using scanned Gaussian, Bessel and array beams. SSPIM provides an accessible toolbox to generate and optimize the desired beam patterns and helps adapting the OpenSPIM system towards a wider range of biological samples.
选择平面照明显微镜 (SPIM) 是动态组织成像的首选方法，因为它结合了高时空分辨率和低光毒性。OpenSPIM 系统旨在为非专业用户提供一种易于使用且灵活的显微镜设置。在这里，我们报告了 Structured SPIM (SSPIM)，它为 OpenSPIM 平台内的光束成形提供了一个开源、用户友好和紧凑的工具包。SSPIM 通过控制空间光调制器 (SLM) 的图案，能够生成各种照明光束的数字图案，包括静态和球形高斯光束、贝塞尔光束和艾里光束。此外，SSPIM 可以为结构照明生成图案，包括非相干和相干阵列光束以及所有类型支持光束的平铺。我们描述了工具包的工作流程，并通过比较各种照明光束的实验数据和模拟结果来演示其应用。最后，使用扫描高斯、贝塞尔和阵列光束对果蝇胚胎进行 3D 成像，研究了 SSPIM 的功能。SSPIM 提供了一个易于使用的工具包，用于生成和优化所需的光束图案，并有助于使 OpenSPIM 系统适应更广泛的生物样本。
