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Centroid and Orientation Precision of Localization Microscopy



Craig D. McGray, Craig R. Copeland, Samuel M. Stavis, Jon C. Geist


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 position, orientation, and motion of microscopic rigid bodies from localization microscopy data. Physical lower bounds on the corresponding 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 through an analysis of the kinematics of a microelectromechanical goniometer indicated by a constellation of fluorescent nanoparticles. Centroid precision and orientation precision as developed here are broadly applicable concepts in optical measurements, due to their generality and the widespread interest in localization microscopy.
Journal of Microscopy


Localization Microscopy, Localization Precision, Centroid Precision, Orientation Precision, microelectromechanical systems, Fluorescence Microscopy


McGray, C. , Copeland, C. , Stavis, S. and Geist, J. (2016), Centroid and Orientation Precision of Localization Microscopy, Journal of Microscopy, [online], (Accessed April 15, 2024)
Created March 11, 2016, Updated November 10, 2018