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Automated tracking of migrating cells in phase-contrast video microscopy sequences using image registration

journal contribution
posted on 12.05.2017, 11:30 authored by A.J. Hand, Tao SunTao Sun, D.C. Barber, D.R. Hose, Sheila MacNeil
Analysis of in vitro cell motility is a useful tool for assessing cellular response to a range of factors. However, the majority of cell-tracking systems available are designed primarily for use with fluorescently labelled images. In this paper, five commonly used tracking systems are examined for their performance compared with the use of a novel in-house celltracking system based on the principles of image registration and optical flow. Image registration is a tool commonly used in medical imaging to correct for the effects of patient motion during imaging procedures and works well on low-contrast images, such as those found in bright-field and phase-contrast microscopy. The five cell-tracking systems examined were Retrac, a manual tracking system used as the gold standard; CellTrack, a recently released freely downloadable software system that uses a combination of tracking methods; ImageJ, which is a freely available piece of software with a plug-in for automated tracking (MTrack2) and Imaris and Volocity, both commercially available automated tracking systems. All systemswere used to track migration of human epithelial cells over ten frames of a phase-contrast time-lapse microscopy sequence. This showed that the in-house image-registration system was the most effective of those tested when tracking non-dividing epithelial cells in low-contrast images, with a successful tracking rate of 95%. The performance of the tracking systems was also evaluated by tracking fluorescently labelled epithelial cells imaged with both phase-contrast and confocal microscopy techniques. The results showed that using fluorescence microscopy instead of phase contrast does improve the tracking efficiency for each of the tested systems. For the in-house software, this improvement was relatively small (<5%difference in tracking success rate),whereasmuch greater improvements in performance were seen when using fluorescence microscopy with Volocity and ImageJ.


We thank EPSRC for providing a PhD studentship for A.J. Hand. The Wellcome Trust (grant no. GR077544AIA) is acknowledged for support of the University of Sheffield Light Microscopy Facility.



  • Aeronautical, Automotive, Chemical and Materials Engineering


  • Chemical Engineering

Published in

Journal of Microscopy






62 - 79


HAND, A.J. ... et al, 2009. Automated tracking of migrating cells in phase-contrast video microscopy sequences using image registration. Journal of Microscopy, 234 (1), pp.62-79


Wiley (Journal © The Royal Microscopical Society, article © the authors)


NA (Not Applicable or Unknown)

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This work is made available according to the conditions of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) licence. Full details of this licence are available at:

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