Synthetic Stochastic Motion Platform for Testing Single Particle Tracking Microscopes

We describe the design and implementation of a control system for testing the performance of single particle tracking microscopes with the method of synthetic motion. Single particle tracking (SPT) has become a common and powerful tool in the study of biomolecular transport in cellular biology, providing the ability to track individual biological macromolecules in their native environment. Existing methods for testing SPT techniques rely on physical simulations and there is a clear need for experimental-based schemes for both comparing different approaches and for characterizing the accuracy and precision of techniques on particular experimental setups. Synthetic motion, that is, using an actuator such as a nanopositioning stage to drive a particle along a known ground-truth trajectory, is a means for achieving these ends. However, the resolution, accuracy, and flexibility of this method is limited by the actuator static and dynamic characteristics. In this work we apply system identification and model inverse feedforward control to increase actuator bandwidth and address some common actuator nonlinearities, develop a set of dimensionless numbers that describe system limitations, and provide a set of guidelines for the practical use of synthetic motion in SPT.