Polarized light microscopy: principles and practice.

Polarized light microscopy: principles and practice.
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偏光显微镜:原理与实践。

DOI:
10.1101/pdb.top078600
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发表时间:
2013
影响因子:
--
通讯作者:
Oldenbourg,Rudolf
Oldenbourg,Rudolf
中科院分区:
--
文献类型:
--
作者:
Oldenbourg,Rudolf

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偏振光显微镜为分析活细胞和组织等不同系统中的分子秩序提供了独特的机会,而不需要使用外源染料或标签。本文简要讨论了偏振光学显微镜的理论,并阐述了使用传统偏振光学显微镜和更专业的偏振显微镜(如LC-PolScope、Oosight或Abrio)的实践。介绍了用于分析光的偏振特性的显微镜组件,如偏振器和补偿器,并讨论了用传统偏光显微镜逐点测量样品双折射的定量技术。新的LC-PolScope极大地提高了该技术的分析能力,为每个图像点同时提供定量的双折射数据,从而以前所未有的灵敏度和光学显微镜的最高分辨率揭示分子有序。讨论的实际方面包括光学元件的选择、样品制备以及将偏振光与差分干涉衬度和荧光显微镜相结合。还包括偏振光学术语的词汇表,以便于讨论使用偏振光学显微镜进行的观测。
Polarized light microscopy provides unique opportunities for analyzing the molecular order in heterogeneous systems, such as living cells and tissues, without using exogenous dyes or labels. This article briefly discusses the theory of polarized light microscopy and elaborates on its practice using a traditional polarized light microscope and more specialized polarization microscopes such as the LC-PolScope, Oosight, or Abrio. The microscope components specific to analyzing the polarization of light, such as polarizer and compensator, are introduced, and quantitative techniques for measuring the birefringence of the specimen point by point using a traditional polarizing microscope are discussed. The new LC-PolScope greatly improves the analytic power of the technique, providing quantitative birefringence data simultaneously for every image point, thereby revealing molecular order with unprecedented sensitivity and at the highest resolution of the light microscope. Practical aspects discussed include the choice of optics, sample preparation, and combining polarized light with differential interference contrast and fluorescence microscopy. A glossary of polarization optical terms is also included to facilitate the discussion of observations made with a polarized light microscope.
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