Quantitative Optical Phase, Amplitude, and Polarization Microscopy

定量光学相位、振幅和偏振显微镜

• 批准号: 7756685
• 负责人: Remy Tumbar
• 金额: $ 18.51万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-10 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7756685
• 关键词: Address; Adherent Culture; Affect; Algorithms; American; Biological; Caenorhabditis elegans; Cells; Chemicals; Clinical Research; Code; Commit; Complex; Computer software; Computers; Coupled; Coupling; Cultured Cells; Custom; Data; Development; Devices; Engineering; Epithelial Cells; Equipment; Fluorescence Microscopy; Funding; Genetic Databases; Genetic Screening; Golgi Apparatus; Histology; Human; Image; Lateral; Lead; Letters; Light; Lighting; Masks; Methods; Microscope; Microscopy; Molecular; Mus; Noise; Optical Methods; Optics; Oral; Organelles; Performance; Phase; Phenotype; Polarization Microscopy; Positioning Attribute; Process; Productivity; Published Comment; Publishing; Research; Research Personnel; Resolution; Resources; Sampling; Series; Specimen; Staging; Staining method; Stains; System; Techniques; Technology; Testing; Tissue Sample; Universities; Work; Writing; career; chemical property; commercialization; cost; digital; editorial; insight; liquid crystal; novel; reconstruction; research study; sensor; tomography

DESCRIPTION (provided by applicant): Proposed research will develop a new quantitative optical microscopy technique providing high resolution, high accuracy, phase, amplitude and polarization images of transparent, unstained, biological specimens. In conjunction with existing methods of quantitative fluorescence microscopy and enabled by its direct and complete access to a target's physical and chemical/molecular information, it will lead to fully digital/computational, comprehensive, multi-modal optical microscopy of biological specimens and significantly advance the capabilities of current optical methods. The new microscope will be built by coupling a novel (demonstrated) optical phase, amplitude and polarization sensor to a modern commercially available microscope chassis. The sensor will be calibrated using a combination of standard techniques (e.g. integrating sphere), extensions of specific techniques used in preliminary work, and custom phase microscopy targets. Imaging performance will be tested on a carefully chosen representative set of transparent specimens. The chosen imaging targets will provide different optical microscopy challenges that are important both for the engineering development and for gaining insight into new ways of solving important biological problems: various monolayer cultured cells, thin mouse tissue samples, and C. elegans worms at different stages of development. Significant improvements to current optical phase microscopy, as enabled by the proposed method, will include: a) imaging (without staining) low contrast organelles, invisible by existing phase contrast or DIC microscopy (e.g. Golgi apparatus); b) vastly more powerful digital processing through consistent application of linear processing algorithms (the data is linear in object information) to quantitative multi-modal imaging data to provide entirely new information/views; and c) true 3D volume information of transparent, unstained, specimens by using the quantitative phase data in diffraction tomography or z-stack reconstruction algorithms. Such improvements will enable, for example, more advanced optical microscopy for high- throughput genetic screening, faster, more accurate (quantitative) automated histology of transparent, unstained, tissue samples, and complex genetic databases containing comprehensive, fully digital, quantitative multi-modal optical microscopy data on specific phenotypes. This research will develop a new quantitative optical microscopy technique capable of providing essential new information to biologists, in a manner amenable to wide use. By providing complete access to the physical/chemical properties of biological specimens, it will enable the application of advanced computer processing techniques to obtain and store (digitally) new information relevant for both basic and clinical research.

描述（由申请人提供）：拟议的研究将开发一种新的定量光学显微镜技术，提供高分辨率，高精度，相位，振幅和偏振图像的透明，未染色，生物标本。与现有的定量荧光显微镜方法结合，并通过直接、完整地获取目标的物理和化学/分子信息，它将实现生物样本的完全数字/计算、全面、多模式光学显微镜，并显着提高当前光学方法的能力。新的显微镜将建立一个新的（演示）光学相位，振幅和偏振传感器耦合到一个现代化的商用显微镜底盘。传感器将使用标准技术（例如积分球）的组合进行校准，在前期工作中使用的特定技术的扩展，以及自定义相位显微镜目标。成像性能将在精心选择的一组代表性透明样本上进行测试。所选择的成像目标将提供不同的光学显微镜挑战，这些挑战对于工程开发和深入了解解决重要生物学问题的新方法都很重要：各种单层培养细胞，薄小鼠组织样品和C。不同发育阶段的蠕虫。通过所提出的方法，对当前光学相位显微镜的显著改进将包括：（未染色）低对比度细胞器，通过现有相差或DIC显微镜不可见（例如高尔基体）; B）通过线性处理算法的一致应用，大大增强了数字处理能力（数据在对象信息中是线性的）到定量多模态成像数据，以提供全新的信息/视图;以及c）通过在衍射层析成像或z-堆叠重建算法中使用定量相位数据获得透明、未染色样本的真实3D体积信息。这样的改进将实现例如用于高通量遗传筛选的更先进的光学显微镜，透明、未染色的组织样品的更快、更准确（定量）的自动化组织学，以及包含关于特定表型的全面、全数字、定量多模态光学显微镜数据的复杂遗传数据库。 这项研究将开发一种新的定量光学显微镜技术，能够以一种适合广泛使用的方式为生物学家提供重要的新信息。通过提供对生物样本的物理/化学特性的完整访问，它将使先进的计算机处理技术的应用能够获得和存储（数字化）与基础和临床研究相关的新信息。