3-D Image Restoration for Polarized Light Microscopy

描述(申请人提供)：我们一直在开发和应用一种新的 一种偏振光学显微镜(P01-Scope)，它显著增强了 传统偏光显微镜的分析能力。P01-范围 集成了一个通用补偿器，由计算机驱动的液晶制成 高灵敏度下测量双折射精细结构的装置(0.02 NM双折射延迟)，高分辨率(0.2 PM)，同时超过 整个视场，以及双折射轴的所有方向 在焦平面内。三维双折射标本，如活体 细胞，用极镜成像为一系列光学切片。每个人 部分包含聚焦结构的双折射，如膜状结构 堆叠、对准的灯丝阵列和单独的灯丝束。然而， 聚焦双折射的测量具有来自外部的虚假贡献 聚焦蜂窝组件。对细部和细部的结构分析 P01-Scope提供的定量信息通常是有限的，因为 这些虚假的双折射贡献。因此，要充分利用 在定量偏振镜图像中，离焦信息需要 已确认并被移除。我们正在提议开发一种计算修复 消除失焦分量对测量影响的程序 聚焦的双折射。P01范围恢复算法将基于 关于为荧光显微镜建立的程序，我们将修改这些程序 说明了偏振部分相干成像的特殊性质 灯。第一个发展目标是建立一个向前程序，即 用于基于测量点扩展函数计算图像特征和 已知或估计的双折射的位置、大小和轴向 物体。第二个目标是找到优化 体积元素中物体双折射的数值估计 载于P01-Scope光学部分。迭代地反过程 将实验图像和计算图像之间的差异降至最低。这一过程 以位置、大小和轴线的稳定三维分布结束 样品双折射的取向。图像恢复程序将 大大增强P01的实用性-适用于生物和医学 应用于非侵入性地分析居住的建筑动态 细胞。

DESCRIPTION (provided by applicant): We have been developing and applying a new type of polarized light microscope (the P01-Scope) which dramatically enhances the analytic power of the traditional polarizing microscope. The P01-Scope incorporates a universal compensator, made from computer driven liquid crystal devices, to measure the birefringent fine structure at high sensitivity (0.02 nm birefringence retardation), high resolution (0.2 pm), simultaneously over the whole field of view, and for all orientations of the birefringence axis within the focus plane. A 3-dimensional birefringent specimen, such as a living cell, is imaged with the Pol-Scope as a series of optical Sections. Each section contains the birefringence of in-focus structures, such as membranous stacks, aligned filament arrays and individual filament bundles. However, the measurements of in-focus birefringences have spurious contributions from out-of focus cellular components. The structural analysis of the detailed and quantitative information afforded by the P01-Scope is often limited because of these spurious birefringence contributions. Therefore, to take full advantage of the quantitative Pol-Scope images, the out-of-focus information needs to be identified and removed. We are proposing to develop a computational restoration procedure to remove the effect of out-of-focus components on the measurements of in-focus birefringences. The P01-Scope restoration algorithm will be based on procedures established for fluorescence microscopy that we will modify to account for the specific properties of partial coherent imaging with polarized light. The first development goal is to establish a forward procedure that is used to compute image features based on measured point spread functions and known or estimated positions, magnitudes and axis orientations of birefringent objects. The second goal is to find the inverse procedure that optimizes a numerical estimate of object birefringences in volume elements that are contained in P01-Scope optical sections. The inverse procedure iteratively minimizes the difference between experimental and computed images. This process ends in a stable 3-D distribution of positions, magnitudes and axis orientations of specimen birefringences. The image restoration procedures will greatly enhance the utility of the P01-Scope for biological and medical applications to non-invasively analyze the architectural dynamics of living cells.