Orientation independent DIC and polarization microscopy

本项目旨在开发和应用新型的差示干涉衬度和偏振光学显微镜 这两种技术的对比度都与样品的取向无关。在第一个授权期内，我们 建立了一种快速变化的与方向无关的差分干涉对比显微镜 剪切方向，不需要任何机械移动的组件。OI-DIC系统接收量化的 具有最高质量的折射率梯度和位相(干质量)图像，这是 任何其他光学显微镜。我们将设计和建造改进的OI-DIC，其光学堆栈包括 DIC棱镜和高质量液晶(LC)单元。由于总厚度较小，此堆叠可 放置在普通显微镜的现有DIC插槽中。新的OI-DIC系统将在两种可见模式下运行 和近红外光谱区，从而打开了识别不同波长结构的可能性- 折射率的依赖关系。我们将开发一种新的与方向无关的偏振显微镜(LC- Polcope)，其仅使用一个LC单元。将DIC和LC-POLSCOPE设备组合到一个单元中将 允许在两种模式之间快速切换，而无需移动任何光学组件 几乎可以同时捕获图像，并且不会出现错位。新系统将产生两个 试件光学薄层的互补像：折射率梯度分布或干态 由结构或固有各向异性引起的双折射的质量和分布。例如，在实时分割中 细胞，OI-DIC图像将清楚地显示染色体，而偏振镜图像将定量描绘 双折射微管在纺锤体中的分布。我们将开发一种方法重建 试件区域折射率的三维分布。这样的能力将对 研究分裂中的活细胞。使用这种新的方法，人们将能够确定 并观察其在分裂过程中的变化。人们也可以数一数 在不扩散或杀死细胞的情况下浓缩的染色体数目。我们将设计和制造新的 视频增强型多色偏振器，能够进行视频速率或更快的图像采集。多彩的 Polcope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 传播是由神经冲动等引起的。

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.