3D imaging of cells and tissues using polarized light microscopy

项目概要 我们建议继续开发偏光显微镜技术来分析 活细胞、组织和整个生物体内部的结构动力学。我们建议添加综合 偏振分析到两种多视图成像技术，双视图平面照明显微镜，或 diSPIM 和光场显微镜。两种类型的偏光显微镜都代表了重要的新成果 首次能够测量三种分子取向参数的进展 高空间（所有三个维度均为 0.5μm）和时间（快于 1Hz）分辨率的维度。的 项目包括捕获多视图所需的硬件、软件和算法的开发 图像数据及其断层扫描重建为双折射和偏振的 3 维图 荧光。仪器开发以多个应用项目为指导，包括改造 成纤维细胞的细胞外胶原基质以及高阶组装中的空间协调 真核细胞内的脓蛋白结构。有关该技术的信息，包括软件组件和 在这笔赠款下开发的仪器设计将提供给用户和开发人员 建立了名为 OpenPolScope.org 的门户网站。

Project Summary We are proposing to continue our development of polarized light microscopy techniques for analyzing the architectural dynamics inside living cells, tissues and whole organisms. We propose to add comprehensive polarization analysis to two multi-view imaging techniques, the dual-view plane illumination microscope, or diSPIM, and the light-field microscope. Both types of polarized light microscopes represent significant new developments that for the first time enable the measurement of molecular orientation parameters in three dimensions at high spatial (0.5µm in all three dimensions) and temporal (faster than 1Hz) resolution. The project includes the development of the hardware, software and algorithms required for capturing multi-view image data and their tomographic reconstruction into 3-dimensional maps of birefringence and polarized fluorescence. The instrument development is guided by several application projects including the remodeling of the extracellular collagen matrix by fibroblast cells and the spatial coordination in the assembly of higher order septin structures inside eukaryotic cells. Information about the technology, including software components and instrument design that are developed under this grant, will be made available for users and developers on an established web portal called OpenPolScope.org.