title: Single-pixel interior filling function approach for detecting and correcting errors in particle tracking.
abstract: We present a general method for detecting and correcting biases in the outputs of particle-tracking experiments. Our approach is based on the histogram of estimated positions within pixels, which we term the single-pixel interior filling function (SPIFF). We use the deviation of the SPIFF from a uniform distribution to test the veracity of tracking analyses from different algorithms. Unbiased SPIFFs correspond to uniform pixel filling, whereas biased ones exhibit pixel locking, in which the estimated particle positions concentrate toward the centers of pixels. Although pixel locking is a well-known phenomenon, we go beyond existing methods to show how the SPIFF can be used to correct errors. The key is that the SPIFF aggregates statistical information from many single-particle images and localizations that are gathered over time or across an ensemble, and this information augments the single-particle data. We explicitly consider two cases that give rise to significant errors in estimated particle locations: undersampling the point spread function due to small emitter size and intensity overlap of proximal objects. In these situations, we show how errors in positions can be corrected essentially completely with little added computational cost. Additional situations and applications to experimental data are explored in SI Appendix In the presence of experimental-like shot noise, the precision of the SPIFF-based correction achieves (and can even exceed) the unbiased Cramér-Rao lower bound. We expect the SPIFF approach to be useful in a wide range of localization applications, including single-molecule imaging and particle tracking, in fields ranging from biology to materials science to astronomy.
authors.affiliation: Department of Physics, Bar-Ilan University, Ramat-Gan 5290002, Israel.|James Franck Institute, The University of Chicago, Chicago, IL 60637.
authors.forename: Stanislav
authors.initials: S
authors.lastname: Burov
authors.affiliation: Department of Chemistry, The University of Chicago, Chicago, IL 60637.
authors.forename: Patrick
authors.initials: P
authors.lastname: Figliozzi
authors.affiliation: James Franck Institute, The University of Chicago, Chicago, IL 60637.|Center for Advanced Radiation Sources, The University of Chicago, Chicago, IL 60637.
authors.forename: Binhua
authors.initials: B
authors.lastname: Lin
authors.affiliation: James Franck Institute, The University of Chicago, Chicago, IL 60637; sarice@uchicago.edu nfschere@uchicago.edu dinner@uchicago.edu.|Department of Chemistry, The University of Chicago, Chicago, IL 60637.
authors.forename: Stuart A
authors.initials: SA
authors.lastname: Rice
authors.affiliation: James Franck Institute, The University of Chicago, Chicago, IL 60637; sarice@uchicago.edu nfschere@uchicago.edu dinner@uchicago.edu.|Department of Chemistry, The University of Chicago, Chicago, IL 60637.|Institute for Biophysical Dynamics, The University of Chicago, Chicago, IL 60637.
authors.forename: Norbert F
authors.initials: NF
authors.lastname: Scherer
authors.affiliation: James Franck Institute, The University of Chicago, Chicago, IL 60637; sarice@uchicago.edu nfschere@uchicago.edu dinner@uchicago.edu.|Department of Chemistry, The University of Chicago, Chicago, IL 60637.|Institute for Biophysical Dynamics, The University of Chicago, Chicago, IL 60637.
authors.forename: Aaron R
authors.initials: AR
authors.lastname: Dinner
keywords: Cramér–Rao lower bound; error correction; imaging; particle tracking; pixel locking