Precision analysis for standard deviation measurements of immobile single fluorescent molecule images.
Precision analysis for standard deviation measurements of immobile single fluorescent molecule images.
复制标题
DOI：
10.1364/oe.18.006563
复制
发表时间：
2010-03-29
期刊：
影响因子：
3.8
通讯作者：
Wang YM
中科院分区：
文献类型：
--
作者：
DeSantis MC;DeCenzo SH;Li JL;Wang YM
固定单个荧光分子强度分布的标准偏差测量对于研究轴向定位、分子取向和荧光成像系统的空间分辨率非常有用。在这里，我们报告使用EMCCD相机成像的单个荧光分子的强度分布的标准偏差测量的精度分析。我们已经开发了一个解析表达式的标准偏差测量误差的一个单一的图像，这是一个函数的总数量的检测到的光子，背景光子噪声，和相机像素大小。理论结果与实验、模拟和数值积分结果吻合较好。使用这个表达式，我们表明，单分子标准偏差测量提供纳米精度的大范围的实验参数。
Standard deviation measurements of intensity profiles of stationary single fluorescent molecules are useful for studying axial localization, molecular orientation, and a fluorescence imaging system’s spatial resolution. Here we report on the analysis of the precision of standard deviation measurements of intensity profiles of single fluorescent molecules imaged using an EMCCD camera. We have developed an analytical expression for the standard deviation measurement error of a single image which is a function of the total number of detected photons, the background photon noise, and the camera pixel size. The theoretical results agree well with the experimental, simulation, and numerical integration results. Using this expression, we show that single-molecule standard deviation measurements offer nanometer precision for a large range of experimental parameters.