超分辨成像已成为活细胞结构和功能成像的关键工具,荧光分子定位是超分辨成像过程中不可缺少的步骤。从超分辨成像角度研究各种荧光分子定位算法性能具有重要的意义。选择5种典型的荧光分子定位算法:质心法、广义质心法、高斯拟合、解线性方程组和极大似然法,以定位精度和定位时间来评价所选择算法的性能。结果表明,1)高斯拟合、极大似然法和广义质心法能高精度对荧光分子定位,不受荧光分子所在子区域提取的影响;2)质心法和解线性方程组法能应用于图像在线分析,但定位精度较低,受子区域提取影响较大;3)当两个荧光分子位于一个衍射斑时,采用这5种算法的定位精度都会急剧下降。 Super-resolution imaging has become a key tool for living cell structure and functional imaging. Fluorescent molecular localization is an indispensable step in super-resolution imaging. It is of great significance to study the performance of various fluorescence molecular localization algorithms from the perspective of super-resolution imaging. Five typical fluorescence molecular localization algorithms were selected: centroid method, generalized centroid method, Gaussian fitting, solution of linear equations and maximum likelihood method, and the performance of the selected algorithm was evaluated by positioning accuracy and positioning time. The results show that 1) Gaussian fitting, maximum likelihood method and generalized centroid method can locate fluorescence molecules with high precision without being affected by the extraction of the fluorescent molecule; 2) The centroid method and the solution of linear equations can be applied to the image On-line analysis, but the positioning accuracy is low, affected by the extraction of the sub-region; 3) When the two fluorescent molecules are located in a diffraction spot, the positioning accuracy of the five algorithms will decline sharply.