Precise particle tracking against a complicated background: polynomial fitting with Gaussian weight
- 1 September 2007
- journal article
- Published by IOP Publishing in Physical Biology
- Vol. 4 (3), 220-227
- https://doi.org/10.1088/1478-3975/4/3/008
Abstract
We present a new particle tracking software algorithm designed to accurately track the motion of low-contrast particles against a background with large variations in light levels. The method is based on a polynomial fit of the intensity around each feature point, weighted by a Gaussian function of the distance from the centre, and is especially suitable for tracking endogeneous particles in the cell, imaged with bright field, phase contrast or fluorescence optical microscopy. Furthermore, the method can simultaneously track particles of all different sizes, and allows significant freedom in their shape. The algorithm is evaluated using the quantitative measures of accuracy and precision of previous authors, using simulated images at variable signal-to-noise ratios. To these we add new tests: the error due to a non-uniform background, and the error due to two particles approaching each other. Finally the tracking of particles in real cell images is demonstrated. The method is made freely available for non-commercial use as a software package with a graphical user-interface, which can be run within the Matlab programming environment.Keywords
This publication has 19 references indexed in Scilit:
- Stepwise Movements in Vesicle Transport of HER2 by Motor Proteins in Living CellsBiophysical Journal, 2007
- Bio-Microrheology: A Frontier in MicrorheologyBiophysical Journal, 2006
- The Role of F-Actin and Myosin in Epithelial Cell RheologyBiophysical Journal, 2006
- Feature point tracking and trajectory analysis for video imaging in cell biologyJournal of Structural Biology, 2005
- Tracking single particles: a user-friendly quantitative evaluationPhysical Biology, 2005
- Microrheology of complex fluidsReports on Progress in Physics, 2005
- Precise Nanometer Localization Analysis for Individual Fluorescent ProbesBiophysical Journal, 2002
- Semiflexible Polymer Network: A View From InsidePhysical Review Letters, 1998
- Particle Tracking Microrheology of Complex FluidsPhysical Review Letters, 1997
- Methods of Digital Video Microscopy for Colloidal StudiesJournal of Colloid and Interface Science, 1996