Fluorescence polarization filtering for accurate single molecule localization
Fluorescence polarization filtering for accurate single molecule localization
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DOI：
10.1063/5.0009904
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发表时间：
2020-06-01
期刊：
影响因子：
5.6
通讯作者：
Enderlein, Joerg
中科院分区：
文献类型：
--
作者：
Nevskyi, Oleksii;Tsukanov, Roman;Enderlein, Joerg
单分子定位显微术（SMLM）已成为超分辨荧光显微术的重要手段之一。它是基于单分子在宽视场显微镜图像中的精确定位。众所周知，如果成像的分子具有固定的取向并且位于接近界面或不完全在显微镜的焦平面内，则定位精度可以显示出显著的偏差。在这封信中，我们提出了一个简单的解决这个问题，这是基于偏振分辨成像。这种方法可以很容易地实现到任何现有的SMLM设置，我们证明了它的性能，通过成像嵌入到聚合物膜，固定其在空间中的方向的单个染料分子。
Single-Molecule Localization Microscopy (SMLM) has become one of the most important methods of super-resolution fluorescence microscopy. It is based on the precise localization of single molecules in wide-field microscopy images. It is well known that the localization accuracy can show a significant bias if the imaged molecules have a fixed orientation and are located either close to an interface or not exactly within the focal plane of the microscope. In this Letter, we propose a simple solution to this problem, which is based on polarization-resolved imaging. This method can be easily implemented into any existing SMLM setup, and we demonstrate its performance by imaging single dye molecules embedded into a polymer film, which fixes their orientation in space.