Online Tracking of Migrating and Proliferating Cells Imaged with Phase-Contrast Microscopy

Automated visual-tracking of cell population in vitro using phase-contrast time-lapse microscopy is vital for the quantitative and systematic study of cell behaviors, including spatiotemporal quantification of migration, proliferation, and apoptosis. The low image quality, high and varying density of the cell culture, and the complexity of cell behaviors pose many challenges to existing tracking techniques. This paper presents a fully-automated multitarget tracking system that can simultaneously track hundreds of cells and efficiently cope with these challenges. The approach exploits a fast topology-constrained level-set method in conjunction with a stochastic motion filter, with a careful formulation that makes it suitable for real-time tracking during acquisition. Our methodology was applied to human tissue cell tracking in vitro under various imaging conditions and yielded a 88.4% tracking accuracy.

[1]  N. Otsu.  A threshold selection method from gray level histograms , 1979 .

[2]  Stanley R. Sternberg,et al.  Biomedical Image Processing , 1983, Computer.

[3]  N. Gordon,et al.  Novel approach to nonlinear/non-Gaussian Bayesian state estimation , 1993 .

[4]  Andrew Blake,et al.  Learning Dynamics of Complex Motions from Image Sequences , 1996, ECCV.

[5]  Dorin Comaniciu,et al.  Real-time tracking of non-rigid objects using mean shift , 2000, Proceedings IEEE Conference on Computer Vision and Pattern Recognition. CVPR 2000 (Cat. No.PR00662).

[6]  T. Başar,et al.  A New Approach to Linear Filtering and Prediction Problems , 2001 .

[7]  Vannary Meas-Yedid,et al.  Segmentation and tracking of migrating cells in videomicroscopy with parametric active contours: a tool for cell-based drug testing , 2002, IEEE Transactions on Medical Imaging.

[8]  J. Sethian,et al.  FRONTS PROPAGATING WITH CURVATURE DEPENDENT SPEED: ALGORITHMS BASED ON HAMILTON-JACOB1 FORMULATIONS , 2003 .

[9]  Xiao Han,et al.  A Topology Preserving Level Set Method for Geometric Deformable Models , 2003, IEEE Trans. Pattern Anal. Mach. Intell..

[10]  Guillermo Sapiro,et al.  Geodesic Active Contours , 1995, International Journal of Computer Vision.

[11]  Scott T. Acton,et al.  Level set analysis for leukocyte detection and tracking , 2004, IEEE Transactions on Image Processing.

[12]  Demetri Terzopoulos,et al.  Snakes: Active contour models , 2004, International Journal of Computer Vision.

[13]  Frank Dellaert,et al.  An MCMC-Based Particle Filter for Tracking Multiple Interacting Targets , 2004, ECCV.

[14]  W. Saltzman.  Tissue Engineering: Engineering Principles for the Design of Replacement Organs and Tissues , 2004 .

[15]  Jean-Christophe Olivo-Marin,et al.  Coupled parametric active contours , 2005, IEEE Transactions on Pattern Analysis and Machine Intelligence.

[16]  Eric D. Miller,et al.  Engineered spatial patterns of FGF-2 immobilized on fibrin direct cell organization. , 2005, Biomaterials.

[17]  Hiroshi Yasaka,et al.  1.3-V/sub pp/ push-pull drive InP Mach-Zehnder modulator module for 40 Gbit/s operation , 2005 .

[18]  W. Clem Karl,et al.  Real-time tracking using level sets , 2005, 2005 IEEE Computer Society Conference on Computer Vision and Pattern Recognition (CVPR'05).

[19]  Philippe Van Ham,et al.  Tracking of migrating cells under phase-contrast video microscopy with combined mean-shift processes , 2005, IEEE Transactions on Medical Imaging.

[20]  Frank Dellaert,et al.  Multitarget tracking with split and merged measurements , 2005, 2005 IEEE Computer Society Conference on Computer Vision and Pattern Recognition (CVPR'05).

[21]  Florent Ségonne,et al.  Segmentation of medical images under topological constraints , 2005 .

[22]  Milan Sonka,et al.  Cell Segmentation, Tracking, and Mitosis Detection Using Temporal Context , 2005, MICCAI.

[23]  Lee E. Weiss,et al.  Bayesian computer-aided experimental design of heterogeneous scaffolds for tissue engineering , 2005, Comput. Aided Des..

[24]  A. Volgenant,et al.  A shortest augmenting path algorithm for dense and sparse linear assignment problems , 1987, Computing.

[25]  Eric D. Miller,et al.  Dose-dependent cell growth in response to concentration modulated patterns of FGF-2 printed on fibrin. , 2006, Biomaterials.

[26]  B. Roysam,et al.  Automated Cell Lineage Construction: A Rapid Method to Analyze Clonal Development Established with Murine Neural Progenitor Cells , 2006, Cell cycle.