Synthetic Stochastic Motion Platform for Testing Single Particle Tracking Microscopes

Real‐Time Feedback‐Driven Single‐Particle Tracking: A Survey and Perspective

“…The final category is a method called synthetic motion. [174,176,177] As with the method of deterministic motion, this approach uses a fluorescent particle fixed to a coverslip and moved by a nanopositioning stage. Instead of a simple, deterministic trajectory, however, the stage is used to realize almost any biologically relevant process, including simple diffusion, tethered motion, confined diffusion, hop diffusion, and more.…”

“…However, the local environment remains simple, without confounding factors such as background fluorescence from nearby structures, and creating accurate realizations of the desired motion, particularly for fast dynamics, can be challenging and is ultimately limited by the capabilities of the nanopositioning stage being used. [177] As RT-FD-SPT microscopes add multi-color detection, contextual imaging, and spectroscopy, it is important to calibrate and characterize each additional function and to ensure the coalignment of each element in the system. A major challenge is that of unintentional motion resulting from thermal expansion, stage drift, and other sources.…”

“…However, the local environment remains simple, without confounding factors such as background fluorescence from nearby structures, and creating accurate realizations of the desired motion, particularly for fast dynamics, can be challenging and is ultimately limited by the capabilities of the nanopositioning stage being used. [ 177 ]…”