Localization accuracy in single-molecule microscopy
Localization accuracy in single-molecule microscopy
复制标题
DOI：
10.1016/s0006-3495(04)74193-4
复制
发表时间：
2004-02-01
影响因子：
3.4
通讯作者：
Ward, ES
中科院分区：
文献类型：
--
作者：
Ober, RJ;Ram, S;Ward, ES
单分子显微镜中最基本的问题之一是确定单个分子位置的准确性。使用Fisher信息矩阵，它示出了对于一个单一的分子的定位精度的极限是由lambda（em）/2 pin（aroot）gammaAt，其中lambda（em），n（a），γ，A，和t表示的发射波长的单一分子，物镜的数值孔径，光学系统的效率，发射速率的单一分子和采集时间，分别。使用蒙特卡罗模拟表明，估计算法可以接近达到表达式中给出的极限。还提供了明确的定量结果，以显示如何减少的限制的定位精度的因素，如检测器的像素和检测系统中的噪声源。结果表明什么是单分子显微镜可以实现的，并为实验设计提供指导方针。
One of the most basic questions in single-molecule microscopy concerns the accuracy with which the location of a single molecule can be determined. Using the Fisher information matrix it is shown that the limit of the localization accuracy for a single molecule is given by lambda(em)/2pin(aroot)gammaAt, where lambda(em), n(a), gamma, A, and t denote the emission wavelength of the single molecule, the numerical aperture of the objective, the efficiency of the optical system, the emission rate of the single molecule and the acquisition time, respectively. Using Monte Carlo simulations it is shown that estimation algorithms can come close to attaining the limit given in the expression. Explicit quantitative results are also provided to show how the limit of the localization accuracy is reduced by factors such as pixelation of the detector and noise sources in the detection system. The results demonstrate what is achievable by single-molecule microscopy and provide guidelines for experimental design.