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Video-Based Tracking of Single Molecules Exhibiting Directed In-Frame Motion

Published online by Cambridge University Press:  30 July 2012

M. Yavuz Yüce ,
Alexandr Jonáš ,
Alper Kiraz  and
Alper T. Erdoğan
M. Yavuz Yüce*
Affiliation:
Department of Physics, Koç University, Istanbul, Turkey
Alexandr Jonáš
Affiliation:
Department of Physics, Koç University, Istanbul, Turkey
Alper Kiraz
Affiliation:
Department of Physics, Koç University, Istanbul, Turkey
Alper T. Erdoğan
Affiliation:
Department of Electrical Engineering, Koç University, Istanbul, Turkey
*
Corresponding author. E-mail: myuce@ku.edu.tr
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Abstract

Trajectories of individual molecules moving within complex environments such as cell cytoplasm and membranes or semiflexible polymer networks provide invaluable information on the organization and dynamics of these systems. However, when such trajectories are obtained from a sequence of microscopy images, they can be distorted due to the fact that the tracked molecule exhibits appreciable directed motion during the single-frame acquisition. We propose a new model of image formation for mobile molecules that takes the linear in-frame motion into account and develop an algorithm based on the maximum likelihood approach for retrieving the position and velocity of the molecules from single-frame data. The position and velocity information obtained from individual frames are further fed into a Kalman filter for interframe tracking of molecules that allows prediction of the trajectory of the molecule and further improves the precision of the position and velocity estimates. We evaluate the performance of our algorithm by calculations of the Cramer-Rao Lower Bound, simulations, and model experiments with a piezo-stage. We demonstrate tracking of molecules moving as fast as 7 pixels/frame (12.6 μm/s) within a mean error of 0.42 pixel (37.43 nm).

Keywords

single molecule microscopysingle molecule trackingmaximum likelihood estimationKalman filtering
Type
Biological Applications: Techniques, Software, and Equipment Development
Information
Microscopy and Microanalysis , Volume 18 , Issue 4 , August 2012 , pp. 781 - 792
Copyright
Copyright © Microscopy Society of America 2012

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