Orientation independent DIC and polarization microscopy

方向无关的 DIC 和偏振显微镜

• 批准号: 8830460
• 负责人: MICHAEL SHRIBAK
• 金额: $ 38.14万
• 依托单位: MARINE BIOLOGICAL LABORATORY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-12-01 至 2017-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8830460
• 关键词: Anisotropy; Behavior; Biological; Birefringence; Cell division; Cells; Cellular Structures; Chromosomes; Code; Color; Computer software; Computers; Data; Dependence; Development; Devices; Elements; Event; Evolution; Generations; Grant; Human Resources; Image; Label; Laboratories; Life; Light Microscope; Measurement; Mechanics; Meiosis; Methods; Microscope; Microtubules; Mitosis; Modification; Molecular; Morphologic artifacts; Motion; Movement; Nerve; Optics; Organelles; Performance; Phase; Polarization Microscopy; Process; Proteins; Refractive Indices; Research Personnel; Resolution; Scientist; Specimen; Staining method; Stains; Structure; System; Techniques; Testing; Thick; Time; Tissues; base; biological research; cell killing; contrast imaging; design; fluorescence imaging; image processing; improved; insight; liquid crystal; miniaturize; molecular assembly/self assembly; novel strategies; particle; polarized light; prototype; reconstruction; time orientation

This project aims to develop and apply new differential interference contrast and polarized light microscope techniques, both of whose contrast is independent of specimen orientation. During the first grant period we built an orientation-independent differential interference contrast (OI-DIC) microscope, which rapidly changes shear directions without any mechanically moved components. The OI-DIC system receives the quantitative refractive index gradient and phase (dry mass) images with the highest quality that cannot be achieved with any other optical microscope. We will design and built the improved OI-DIC with an optical stack consisting of DIC prisms and high quality liquid crystal (LC) cells. Because of the small total thickness, this stack can be placed into an existing DIC slot of a regular microscope. The new OI-DIC system will operate in both the visible and near- IR spectral regions, thus opening up the possibility of identifying structures with different wavelength- dependence of refractive indices. We will develop a new orientation-independent polarization microscope (LC- polscope), which employs only one LC cell. The DIC and LC-polscope devices combined into one unit will allow rapid switching between the two modes without the need to move any optical components so that both images can be captured nearly simultaneously and without misalignment. The new system will yield two complementary images of thin optical section of the specimen: distribution of refractive index gradient or dry mass and distribution of birefringence due to structural or intrinsic anisotropy. For example, in a live dividing cell, the OI-DIC image will clearly show the chromosomes while the polscope image will quantitatively depict the distribution of the birefringent microtubules in the spindle. We will develop a method for reconstruction the 3D distributions of refractive index of specimen regions. Such capabilities will be especially important for studying live cells in division. With the new approach, one will be able to determine the dry mass of chromosomes before division and observe its change during the division process. One could also count the number of condensed chromosomes without spreading or killing the cell. We will design and fabricate new video-enhanced polychromatic polscope, capable of video-rate or faster image acquisition. The polychromatic polscope will be able to provide sharp images of fast moving birefringent organelles, rapid birefringence propagation caused be nerve impulses, etc.

本项目旨在研制和应用新型微分干涉衬度和偏振光显微镜 技术，其对比度是独立的试样取向。在第一个阶段，我们 建立了一个方向无关的微分干涉对比（OI-DIC）显微镜，它迅速变化， 剪切方向，没有任何机械移动的组件。OI-DIC系统接收定量的 折射率梯度和相位（干质量）图像具有最高质量， 其他光学显微镜我们将设计和构建改进的OI-DIC，其光学堆栈包括 DIC棱镜和高品质液晶（LC）单元。由于小的总厚度，该叠层可以是 放置在常规显微镜的现有DIC槽中。新的OI-DIC系统将在可见光 和近红外光谱区域，从而打开了识别不同波长结构的可能性- 折射率的依赖性。我们将研制一种新的与取向无关的偏振显微镜（LC- polscope），其仅采用一个LC单元。DIC和LC polscope设备组合成一个单元， 允许在两种模式之间快速切换而不需要移动任何光学部件 可以几乎同时捕获图像并且没有未对准。新系统将产生两个 样品薄光学切片的互补图像：折射率梯度或干燥的分布 由于结构或固有的各向异性，双折射的质量和分布。例如，在一个活的分割中， 细胞，OI-DIC图像将清楚地显示染色体，而polscope图像将定量描绘 纺锤体中双折射微管的分布。我们将开发一种方法来重建 样品区域折射率的三维分布。这种能力对以下方面尤其重要： 研究活细胞的分裂过程有了新的方法，人们将能够确定的干质量 观察其在分裂过程中的变化。人们也可以把 浓缩的染色体数量，而不扩散或杀死细胞。我们将设计和制造新的 视频增强多色偏光镜，能够以视频速率或更快的速度采集图像。多色 偏振镜将能够提供快速移动的双折射细胞器的清晰图像，快速双折射 由神经冲动等引起的传播。