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We describe how a single-particle tracking experiment should be designed inorder for its recorded trajectories to contain the most information about atracked particle's diffusion coefficient. The precision of estimators for thediffusion coefficient is affected by motion blur, limited photon statistics,and the length of recorded time-series. We demonstrate for a particleundergoing free diffusion that precision is negligibly affected by motion blurin typical experiments, while optimizing photon counts and the number ofrecorded frames is the key to precision. Building on these results, we describefor a wide range of experimental scenarios how to choose experimentalparameters in order to optimize the precision. Generally, one should choosequantity over quality: experiments should be designed to maximize the number offrames recorded in a time-series, even if this means lower information contentin individual frames.

Optimizing experimental parameters for tracking of diffusing particles