3-D Image Restoration for Polarized Light Microscopy

偏光显微镜的 3D 图像恢复

• 批准号: 6526004
• 负责人: RUDOLF OLDENBOURG
• 金额: $ 23万
• 依托单位: MARINE BIOLOGICAL LABORATORY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-15 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6526004
• 关键词: automated data processing; bioengineering /biomedical engineering; bioimaging /biomedical imaging; birefringences; cell component structure /function; computer program /software; image processing; light microscopy; mathematics; optical polarization; structural biology

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.

描述（由申请人提供）：我们一直在开发和应用一种新的 类型的偏光显微镜（P01-Scope）可显着增强 传统偏光显微镜的分析能力。 P01-范围 包含一个通用补偿器，由计算机驱动的液晶制成 设备，以高灵敏度测量双折射精细结构（0.02 nm 双折射延迟），高分辨率（0.2 pm），同时超过 整个视野以及双折射轴的所有方向 在焦平面内。 3 维双折射样本，例如活体 使用 Pol-Scope 将细胞成像为一系列光学切片。每个 部分包含焦点结构的双折射，例如膜 堆叠、对齐的灯丝阵列和单独的灯丝束。然而， 焦点内双折射的测量存在来自外的杂散贡献 聚焦细胞成分。详细的结构分析 P01-Scope 提供的定量信息通常是有限的，因为 这些寄生双折射的贡献。因此，要充分利用 在定量 Pol-Scope 图像中，需要将离焦信息 识别并删除。我们建议开发一种计算恢复方法 消除失焦组件对测量影响的程序 聚焦双折射。 P01-Scope 恢复算法将基于 我们将修改为荧光显微镜建立的程序 考虑偏振部分相干成像的特定属性 光。第一个发展目标是建立一个前向程序 用于根据测量的点扩散函数计算图像特征和 双折射的已知或估计位置、大小和轴方向 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-Scope 光学部分中。迭代逆过程 最大限度地减少实验图像和计算图像之间的差异。这个过程 最终形成位置、幅度和轴的稳定 3D 分布 样本双折射的方向。图像恢复程序将 大大增强了 P01-Scope 在生物和医学领域的实用性 用于非侵入性分析生活建筑动态的应用程序 细胞。