基于单分子定位的超分辨显微中荧光激发密度的优化  

The Optimization of Fluorescence Imaging Density in Single-Molecule Localization Based Super-Resolution Microscopy

作　　者：杨光[1,2,3] 熊大曦[1] 梁永[1] 李辉[1] 

机构地区：[1]中国科学院苏州生物医学工程技术研究所,江苏省医用光学重点实验室,江苏苏州215163 [2]中国科学院长春光学精密机械与物理研究所,长春130033 [3]中国科学院大学,北京100039

出　　处：《生物物理学报》2014年第5期380-390,共11页Acta Biophysica Sinica

基　　金：国家自然科学基金项目(51376191)~~

摘　　要：基于单分子定位的超分辨显微技术中,荧光点的中心位置可通过对每个荧光点进行单点扩散函数(point spread function,PSF)逐个拟合定位或对多个荧光点进行多PSF同时拟合定位获得。定位误差以及图像采样时间与荧光激发密度直接相关。为定量得到基于这两种算法的最优荧光激发密度,模拟了成像与定位过程,比较了常见的几种荧光分子在采用基于单PSF逐个拟合和多PSF同时拟合两种算法定位时的中心定位误差、荧光获取比率、获取的有效荧光点数与荧光激发密度间的关系,进而得到了采用这两种算法对不同荧光染剂进行超分辨成像时的最优荧光激发密度。该结果对单分子成像过程中荧光激发密度的选择和激发激光的强度控制具有指导意义。In single molecule localization based supper resolution microscopy, center position of activated fluorescent marker can be localized by fitting with single point spread function（PSF） or multiple PSFs. The localization error and the total sampling time of the image are dependent on the fluorescence molecule imaging density. To obtain the best imaging density quantitatively, the authors simulated the super-resolution imaging process and localized marker positions with single or multiple PSFs. The two algorithms were compared with regard to localization error, recall ratio and number of valid molecules. At last, the best imaging densities corresponding to different molecules and algorithms were obtained. The results are instructive for the choice of imaging density and the control of activation laser intensity.

关 键 词：显微 荧光 单分子定位 激发密度 

分 类 号：Q-334[生物学]