Zukunftskolleg, University of Konstanz, Konstanz, Germany.
Phys Chem Chem Phys. 2013 May 7;15(17):6196-205. doi: 10.1039/c3cp44693j.
Single molecule tracking provides unprecedented insights into diffusional processes of systems in life and material sciences. Determination of molecule positions with high accuracy and correct connection of the determined positions to tracks is a challenging task with, so far, no universal solution for single fluorescing molecules tackling the challenge of low signal-to-noise ratios, frequent blinking and photo bleaching. Thus, the development of novel algorithms for automatic single molecule fluorescence tracking is essential to analyse the huge amount of diffusional data obtained with single molecule widefield fluorescence microscopy. Here, we present a novel tracking model using a top-down polyhedral approach which can be implemented effectively using standard linear programming solvers. The results of our tracking approach are compared to the ground truth of simulated data with different diffusion coefficients, signal-to-noise ratios and particle densities. We also determine the dependency of blinking on the analysed distribution of diffusion coefficients. To confirm the functionality of our tracking method, the results of automatic tracking and manual tracking by a human expert are compared and discussed.
单分子追踪技术为生命科学和材料科学领域的扩散过程提供了前所未有的深入见解。以高精度确定分子位置，并将确定的位置正确地连接到轨迹上，是一项具有挑战性的任务，到目前为止，还没有针对单个荧光分子的通用解决方案能够应对低信噪比、频繁闪烁和光漂白的挑战。因此，开发用于自动单分子荧光追踪的新算法对于分析使用单分子宽场荧光显微镜获得的大量扩散数据至关重要。在这里，我们提出了一种使用自上而下的多面体方法的新追踪模型，该模型可以有效地使用标准线性规划求解器来实现。我们的追踪方法的结果与具有不同扩散系数、信噪比和粒子密度的模拟数据的真实情况进行了比较。我们还确定了闪烁对分析扩散系数分布的依赖性。为了确认我们的追踪方法的功能，我们比较和讨论了自动追踪和人类专家手动追踪的结果。
