Centroid precision and orientation precision of planar localization microscopy

The concept of localization precision, which is essential to localization microscopy, is formally extended from optical point sources to microscopic rigid bodies. Measurement functions are presented to calculate the planar pose and motion of microscopic rigid bodies from localization microscopy data. Physical lower bounds on the associated uncertainties – termed centroid precision and orientation precision – are derived analytically in terms of the characteristics of the optical measurement system and validated numerically by Monte Carlo simulations. The practical utility of these expressions is demonstrated experimentally by an analysis of the motion of a microelectromechanical goniometer indicated by a sparse constellation of fluorescent nanoparticles. Centroid precision and orientation precision, as developed here, are useful concepts due to the generality of the expressions and the widespread interest in localization microscopy for super‐resolution imaging and particle tracking.