Motion blur filtering: A statistical approach for extracting confinement forces and diffusivity from a single blurred trajectory.

Single particle tracking (SPT) can aid in understanding a variety of complex spatiotemporal processes. However, quantifying diffusivity and confinement forces from individual live cell trajectories is complicated by inter- and intratrajectory kinetic heterogeneity, thermal fluctuations, and (experimentally resolvable) statistical temporal dependence inherent to the underlying molecule's time correlated confined dynamics experienced in the cell. The problem is further complicated by experimental artifacts such as localization uncertainty and motion blur. The latter is caused by the tagged molecule emitting photons at different spatial positions during the exposure time of a single frame. The aforementioned experimental artifacts induce spurious time correlations in measured SPT time series that obscure the information of interest (e.g., confinement forces and diffusivity). We develop a maximum likelihood estimation (MLE) technique that decouples the above noise sources and systematically treats temporal correlation via time series methods. This ultimately permits a reliable algorithm for extracting diffusivity and effective forces in confined or unconfined environments. We illustrate how our approach avoids complications inherent to mean square displacement or autocorrelation techniques. Our algorithm modifies the established Kalman filter (which does not handle motion blur artifacts) to provide a likelihood based time series estimation procedure. The result extends A. J. Berglund's motion blur model [Phys. Rev. E 82, 011917 (2010)PLEEE81539-375510.1103/PhysRevE.82.011917] to handle confined dynamics. The approach can also systematically utilize (possibly time dependent) localization uncertainty estimates afforded by image analysis if available. This technique, which explicitly treats confinement and motion blur within a time domain MLE framework, uses an exact likelihood (time domain methods facilitate analyzing nonstationary signals). Our estimator is demonstrated to be consistent over a wide range of exposure times (5 to 100 ms), diffusion coefficients (1×10^{-3} to 1μm^{2}/s), and confinement widths (100 nm to 2μm). We demonstrate that neglecting motion blur or confinement can substantially bias estimation of kinetic parameters of interest to researchers. The technique also permits one to check statistical model assumptions against measured individual trajectories without "ground truth." The ability to reliably and consistently extract motion parameters in trajectories exhibiting confined and/or non-stationary dynamics, without exposure time artifacts corrupting estimates, is expected to aid in directly comparing trajectories obtained from different experiments or imaging modalities. A Python implementation is provided (open-source code will be maintained on GitHub; see also the Supplemental Material with this paper).