Laboratory of Biophysics, Wageningen University and Research, Stippeneng 4, 6708 WE Wageningen, The Netherlands.
Faculty of Applied Sciences, Delft University of Technology, Lorentzweg 1, 2628 CJ Delft, The Netherlands.
J Chem Phys. 2018 Mar 28;148(12):123311. doi: 10.1063/1.5005899.
We present a fast and model-free 2D and 3D single-molecule localization algorithm that allows more than 3 × 10 localizations per second to be calculated on a standard multi-core central processing unit with localization accuracies in line with the most accurate algorithms currently available. Our algorithm converts the region of interest around a point spread function to two phase vectors (phasors) by calculating the first Fourier coefficients in both the x- and y-direction. The angles of these phasors are used to localize the center of the single fluorescent emitter, and the ratio of the magnitudes of the two phasors is a measure for astigmatism, which can be used to obtain depth information (z-direction). Our approach can be used both as a stand-alone algorithm for maximizing localization speed and as a first estimator for more time consuming iterative algorithms.
我们提出了一种快速且无模型的 2D 和 3D 单分子定位算法,它允许在标准多核中央处理器上每秒计算超过 3×10 次的定位,并且具有与当前最准确算法相当的定位精度。我们的算法通过计算 x 和 y 方向上的前两个傅里叶系数,将点扩散函数周围的感兴趣区域转换为两个相位向量(phasors)。这些phasors 的角度用于定位单荧光发射器的中心,并且两个phasors 的幅度比是彗差的度量,可以用于获得深度信息(z 方向)。我们的方法既可以作为最大化定位速度的独立算法,也可以作为更耗时的迭代算法的初始估计器。