Yoon Ji Won, Bruckbauer Andreas, Fitzgerald William J, Klenerman David
Department of Engineering and Department of Chemistry, University of Cambridge, Cambridge, United Kingdom.
Biophys J. 2008 Jun;94(12):4932-47. doi: 10.1529/biophysj.107.116285. Epub 2008 Mar 13.
Single molecule tracking is widely used to monitor the change in position of lipids and proteins in living cells. In many experiments in which molecules are tagged with a single or small number of fluorophores, the signal/noise ratio may be limiting, the number of molecules is not known, and fluorophore blinking and photobleaching can occur. All these factors make accurate tracking over long trajectories difficult and hence there is still a pressing need to develop better algorithms to extract the maximum information from a sequence of fluorescence images. We describe here a Bayesian-based inference approach, based on a trans-dimensional sequential Monte Carlo method that utilizes both the spatial and temporal information present in the image sequences. We show, using model data, where the real trajectory of the molecule is known, that our method allows accurate tracking of molecules over long trajectories even with low signal/noise ratio and in the presence of fluorescence blinking and photobleaching. The method is then applied to real experimental data.
单分子追踪被广泛用于监测活细胞中脂质和蛋白质的位置变化。在许多用单个或少量荧光团标记分子的实验中,信噪比可能受限,分子数量未知,并且荧光团可能会发生闪烁和光漂白。所有这些因素使得在长轨迹上进行精确追踪变得困难,因此仍然迫切需要开发更好的算法,以便从一系列荧光图像中提取最大信息。我们在此描述一种基于贝叶斯的推理方法,该方法基于跨维度序贯蒙特卡罗方法,利用图像序列中存在的空间和时间信息。我们使用已知分子真实轨迹的模型数据表明,即使在低信噪比以及存在荧光闪烁和光漂白的情况下,我们的方法也能在长轨迹上精确追踪分子。然后将该方法应用于实际实验数据。