论文标题
静态单分子定位显微镜图像的横向分辨率度量，该图像来自时间分辨的对相关函数
A lateral resolution metric for static single molecule localization microscopy images from time-resolved pair correlation functions
论文作者
论文摘要
单分子定位显微镜（SMLM）允许可视化细胞结构的数量级小于可见光的衍射极限，并且对SMLM数据集的分辨率进行准确，客观评估是图像处理和分析管道的重要方面。在这里，我们提出了一种简单的方法，该方法使用对在空间和时间上评估的配对自相关函数来测量静态样品的SMLM图像的时间间隔依赖点传播函数。使用这种方法，我们证明，由于漂移和漂移校正算法引起的额外不确定性，实验获得的图像通常具有比单独定位精度所预期的有效点扩散函数。该分辨率度量报告了如何精确地测量标记对象之间的成对距离，并与常用的傅立叶环相关度度量互补，该指标也考虑了空间采样。该方法在模拟的局部定位，DNA折纸统治者和由染料偶联抗体或荧光融合蛋白标记的细胞结构中得到了证明。
Single molecule localization microscopy (SMLM) permits the visualization of cellular structures an order of magnitude smaller than the diffraction limit of visible light, and an accurate, objective evaluation of the resolution of an SMLM dataset is an essential aspect of the image processing and analysis pipeline. Here we present a simple method that uses the pair autocorrelation function evaluated both in space and time to measure the time-interval dependent point-spread function of SMLM images of static samples. Using this approach, we demonstrate that experimentally obtained images typically have effective point spread functions that are broader than expected from the localization precision alone, due to additional uncertainty arising from drift and drift correction algorithms. This resolution metric reports on how precisely one can measure pairwise distances between labeled objects and is complementary to the commonly used Fourier Ring Correlation metric that also considers spatial sampling. The method is demonstrated on simulated localizations, DNA origami rulers, and cellular structures labelled by dye-conjugated antibodies or fluorescent fusion proteins.
