Gu Kai, Liu Chunming
School of Polymer Science and Polymer Engineering, University of Akron, Akron, Ohio 44325, United States.
Department of Chemistry, University of Akron, Akron, Ohio 44325, United States.
Chem Biomed Imaging. 2023 Jun 6;1(3):234-241. doi: 10.1021/cbmi.3c00043. eCollection 2023 Jun 26.
Single-molecule localization microscopy circumvents the diffraction limit of traditional fluorescence microscopy by detecting the photoemission signals of individual fluorescent molecules. The accurate recognitions of fluorescence molecules/events are critical to single-molecule/super-resolution imaging experiments, which determine the precision of molecular localizations and the quality of the image reconstruction. Herein, we presented a single-molecule detection method which relied on the temporal pixel intensity fluctuation. The method was capable of quickly determining the approximate localizations of fluorescence events with high sensitivity. We evaluated the performance of the method under a series of signal-to-noise ratios (SNR) and discussed the criterion of setting the temporal fluctuation threshold to achieve the optimized spots recognition results.
单分子定位显微镜通过检测单个荧光分子的光发射信号，规避了传统荧光显微镜的衍射极限。荧光分子/事件的准确识别对于单分子/超分辨率成像实验至关重要，这些实验决定了分子定位的精度和图像重建的质量。在此，我们提出了一种基于时间像素强度波动的单分子检测方法。该方法能够以高灵敏度快速确定荧光事件的近似定位。我们评估了该方法在一系列信噪比（SNR）下的性能，并讨论了设置时间波动阈值以获得优化的光斑识别结果的标准。
