3-D Image Restoration for Polarized Light Microscopy

描述（由申请人提供）：我们一直在开发和应用一种新的 一种偏振光显微镜（P01-Scope）， 传统偏光显微镜的分析能力。P01-范围 包含一个通用的补偿器，由计算机驱动的液晶 器件，以高灵敏度（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，是成像与Pol-Scope作为一系列的光学部分。每个 部分包含聚焦结构的双折射，例如膜 堆叠、对齐的细丝阵列和单独的细丝束。但 聚焦双折射的测量具有来自非聚焦双折射的虚假贡献。 聚焦细胞成分。详细的结构分析， P01范围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，为了充分利用 在定量Pol-Scope图像中，需要将失焦信息 识别并删除。我们建议开发一种计算恢复 消除离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 我们将修改荧光显微镜建立的程序， 解释了具有偏振的部分相干成像的特定性质， 光第一个开发目标是建立一个向前的程序， 用于基于测量的点扩散函数计算图像特征， 已知或估计的双折射的位置、幅度和轴取向 对象第二个目标是找到优化的逆过程 体积元素中物体双折射的数值估计， 包含在P01-内窥镜光学部分中。迭代的逆过程 最小化实验图像和计算图像之间的差异。这个过程 以稳定的位置、大小和轴的三维分布结束 试样双折射的方向。图像恢复程序将 大大提高了P01-Scope在生物和医疗领域的实用性 非侵入性分析生活建筑动态的应用程序 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.