Department of Biomedical Engineering, Southern University of Science and Technology, Shenzhen, 518055, China.
European Molecular Biology Laboratory, Cell Biology and Biophysics, 69117, Heidelberg, Germany.
Nat Commun. 2022 Jun 6;13(1):3133. doi: 10.1038/s41467-022-30719-4.
Multi-channel detection in single-molecule localization microscopy greatly increases information content for various biological applications. Here, we present globLoc, a graphics processing unit based global fitting algorithm with flexible PSF modeling and parameter sharing, to extract maximum information from multi-channel single molecule data. As signals in multi-channel data are highly correlated, globLoc links parameters such as 3D coordinates or photon counts across channels, improving localization precision and robustness. We show, both in simulations and experiments, that global fitting can substantially improve the 3D localization precision for biplane and 4Pi single-molecule localization microscopy and color assignment for ratiometric multicolor imaging.
多通道单分子定位显微镜检测极大地增加了各种生物应用的信息量。在这里，我们提出了 globLoc，这是一种基于图形处理单元的全局拟合算法，具有灵活的 PSF 建模和参数共享功能，可从多通道单分子数据中提取最大信息。由于多通道数据中的信号高度相关，globLoc 链接了跨通道的参数，如 3D 坐标或光子计数，从而提高了定位精度和鲁棒性。我们通过模拟和实验表明，全局拟合可以显著提高双平面和 4Pi 单分子定位显微镜的 3D 定位精度，以及比率多色成像的颜色分配。
