Various techniques have been developed to measure the 2D and 3D positions and 2D and 3D orientations of fluorescent molecules with improved precision over standard epifluorescence microscopes. Due to the challenging signal-to-background ratio in typical single-molecule experiments, it is essential to choose an imaging system optimized for the specific target sample. In this work, we compare the performance of multiple state-of-the-art and commonly used methods for orientation localization microscopy against the fundamental limits of measurement precision. Our analysis reveals optimal imaging methods for various experiment conditions and sample geometries. Interestingly, simple modifications to the standard fluorescence microscope exhibit superior performance in many imaging scenarios.

中文翻译：

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单分子取向定位显微镜II：性能比较

已经开发了各种技术来测量荧光分子的2D和3D位置以及2D和3D方向，其精度比标准落射荧光显微镜高。由于典型的单分子实验中具有极高的信噪比，因此必须选择针对特定目标样品进行优化的成像系统。在这项工作中，我们将测量定位显微镜的多种最新技术和常用方法的性能与测量精度的基本极限进行了比较。我们的分析揭示了针对各种实验条件和样品几何形状的最佳成像方法。有趣的是，对标准荧光显微镜的简单修改在许多成像场景中都表现出卓越的性能。