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Three-dimensional orientation measurements of symmetric single chromophores using polarization microscopy

机译：使用偏振显微镜对对称单发色团的三维取向测量

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A complete understanding of any complex molecular system generally requires a knowledge of the three-dimensional (3D) orientation of its components relative both to each other, and to directional perturbations such as interfaces and electromagnetic fields. Far-field polarization microscopy is a convenient and widespread technique for detecting and measuring the orientation of single chromophores. But because the polarized electromagnetic field that is used to probe the system lacks a significant longitudinal component, it was thought that, in general, only 2D orientation information could be obtained. Here we demonstrate that far-field polarization microscopy can yield the 3D orientation of certain highly symmetric single chromophores (CdSe nanocrystal quantum dots in the present case). The key requirement is that the chromophores must have a degenerate transition dipole oriented isotropically in two dimensions, which gives rise to a perpendicular 'dark axis' that does not couple to the light field. By measuring the fluorescence intensity from the dipole as a function of polarization angle, it is possible to calculate both the tilt angle between the dark axis and the sample plane, as well as the in-plane orientation, and hence obtain the 3D orientation of the chromophore
机译：对任何复杂分子系统的完整理解通常都需要了解其组件相对于彼此以及方向性扰动（例如界面和电磁场）的三维（3D）方向。远场偏振显微镜是一种方便且广泛使用的技术，用于检测和测量单个发色团的方向。但是，由于用于探测系统的极化电磁场缺少明显的纵向分量，因此人们认为通常只能获得2D方向信息。在这里，我们证明远场偏振显微镜可以产生某些高度对称的单发色团（在当前情况下为CdSe纳米晶体量子点）的3D方向。关键要求是，生色团必须具有在二维上各向同性取向的简并过渡偶极子，这将导致不与光场耦合的垂直“暗轴”。通过测量来自偶极子的荧光强度作为偏振角的函数，可以计算暗轴和样品平面之间的倾斜角以及面内方向，从而获得其3D方向。发色团

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