How Easy Is It to Learn Motion Models from Widefield Fluorescence Single Particle Tracks?

Abstract: Motion models are often deduced from fluorescence widefield tracking experiments by analyzing single-particle trajectories post-processed from the data. This analysis immediately raises the following question: To what degree is our ability to learn motion models impacted by analyzing post-processed trajectories versus the raw measurements? To answer this question, we mathematically formulate a data likelihood for diffraction-limited fluorescence widefield tracking experiments. In particular, we explicitly make the likelihood's dependence on the motion model versus the emission model (or measurement model). The emission model describes how photons emitted by fluorescently labeled particles are distributed in space according to the optical point spread function, with intensities subsequently integrated over a pixel, and convoluted with camera noise. Logic dictates that if the data likelihood is primarily informed by the motion model, then it should be straightforward to learn the motion model from the trajectory post-processed from the data. On the other hand, if the majority of the likelihood is numerically dominated by the emission model, then the post-processed trajectory inferred from data is primarily informed by the emission model, and very little information on the motion model permeates into the post-processed trajectories analyzed downstream to learn motion models. We find that for typical diffraction-limited fluorescence experiments, the emission model often robustly contributes approximately 99% to the likelihood, leaving motion models to explain approximately 1% of the data. This result immediately casts doubt on our ability to reliably learn motion models from post-processed data, raising further questions on the significance of motion models learned thus far from post-processed single-particle trajectories from single-molecule widefield fluorescence tracking experiments.