3D imaging of cells and tissues using polarized light microscopy

项目摘要 我们建议继续开发偏振光显微镜技术，用于分析 活细胞、组织和整个有机体内的建筑动力学。我们建议增加全面的 偏振分析两种多视点成像技术，双视点平面照明显微镜，或 DISPIM和光场显微镜。这两种类型的偏振光显微镜都代表着重大的新技术 首次使分子取向参数能够在三年内测量的发展 高空间分辨率(所有三个维度均为0.5微米)和时间分辨率(快于1赫兹)。这个 该项目包括开发捕获多视角所需的硬件、软件和算法 图像数据及其层析重建成双折射和偏振的三维地图 荧光。该仪器的开发是由几个应用项目指导的，包括 成纤维细胞形成的细胞外胶原基质及其在高阶组装中的空间配位 真核细胞内的Septin结构。有关该技术的信息，包括软件组件和 在这项资助下开发的仪器设计将在 建立了名为OpenPolScope e.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.