Advanced Level-Set-Based Cell Tracking in Time-Lapse Fluorescence Microscopy

Cell segmentation and tracking in time-lapse fluorescence microscopy images is a task of fundamental importance in many biological studies on cell migration and proliferation. In recent years, level sets have been shown to provide a very appropriate framework for this purpose, as they are well suited to capture topological changes occurring during mitosis, and they easily extend to higher dimensional image data. This model evolution approach has also been extended to deal with many cells concurrently. Notwithstanding its high potential, the multiple-level-set method suffers from a number of shortcomings, which limit its applicability to a larger variety of cell biological imaging studies. In this paper, we propose several modifications and extensions to the coupled-active-surfaces algorithm, which considerably improve its robustness and applicability. Our algorithm was validated by comparing it to the original algorithm and two other cell segmentation algorithms. For the evaluation, four real fluorescence microscopy image datasets were used, involving different cell types and labelings that are representative of a large range of biological experiments. Improved tracking performance in terms of precision (up to 11%), recall (up to 8%), ability to correctly capture all cell division events, and computation time (up to nine times reduction) is achieved.

[1]  Vladimir Kolmogorov,et al.  Computing geodesics and minimal surfaces via graph cuts , 2003, Proceedings Ninth IEEE International Conference on Computer Vision.

[2]  Pavel Vesely,et al.  Handbook of Biological Confocal Microscopy, 3rd ed. By James B. Pawley, Editor. Springer Science + Business Media, LLC, New York (2006). ISBN 10: 0‐387‐25921‐X; ISBN 13: 987‐0387‐25921‐5; hardback; 28 + 985 pages , 2007 .

[3]  R. Eils,et al.  Computational imaging in cell biology , 2003, The Journal of cell biology.

[4]  Ross T. Whitaker,et al.  A Streaming Narrow-Band Algorithm: Interactive Computation and Visualization of Level Sets , 2004, IEEE Trans. Vis. Comput. Graph..

[5]  Christophe Zimmer,et al.  Segmenting and tracking fluorescent cells in dynamic 3-D microscopy with coupled active surfaces , 2005, IEEE Transactions on Image Processing.

[6]  Bülent Sankur,et al.  Survey over image thresholding techniques and quantitative performance evaluation , 2004, J. Electronic Imaging.

[7]  W. V. van Cappellen,et al.  Dynamics of relative chromosome position during the cell cycle. , 2004, Molecular biology of the cell.

[8]  Marina Chicurel,et al.  Cell Migration Research Is on the Move , 2002, Science.

[9]  Ioannis Pitas,et al.  Automated evaluation of her-2/neu status in breast tissue from fluorescent in situ hybridization images , 2005, IEEE Transactions on Image Processing.

[10]  Wiro J. Niessen,et al.  A VARIATIONAL MODEL FOR LEVEL-SET BASED CELL TRACKING IN TIME-LAPSE FLUORESCENCE MICROSCOPY IMAGES , 2007, 2007 4th IEEE International Symposium on Biomedical Imaging: From Nano to Macro.

[11]  Tony F. Chan,et al.  Active contours without edges , 2001, IEEE Trans. Image Process..

[12]  J. Pawley,et al.  Handbook of Biological Confocal Microscopy , 1990, Springer US.

[13]  Wiro J. Niessen,et al.  A new detection scheme for multiple object tracking in fluorescence microscopy by joint probabilistic data association filtering , 2008, 2008 5th IEEE International Symposium on Biomedical Imaging: From Nano to Macro.

[14]  Frank Dellaert,et al.  MCMC-based particle filtering for tracking a variable number of interacting targets , 2005, IEEE Transactions on Pattern Analysis and Machine Intelligence.

[15]  Xiaobo Zhou,et al.  Nuclei Segmentation Using Marker-Controlled Watershed, Tracking Using Mean-Shift, and Kalman Filter in Time-Lapse Microscopy , 2006, IEEE Transactions on Circuits and Systems I: Regular Papers.

[16]  H. Erfle,et al.  High-throughput RNAi screening by time-lapse imaging of live human cells , 2006, Nature Methods.

[17]  R. Deriche,et al.  A variational framework for active and adaptative segmentation of vector valued images , 2002, Workshop on Motion and Video Computing, 2002. Proceedings..

[18]  K. R. Ramakrishnan,et al.  Stability and convergence of the level set method in computer vision , 2007, Pattern Recognit. Lett..

[19]  Stephen T. C. Wong,et al.  High content image analysis for human H4 neuroglioma cells exposed to CuO nanoparticles , 2007, BMC biotechnology.

[20]  S. Gibson,et al.  Experimental test of an analytical model of aberration in an oil-immersion objective lens used in three-dimensional light microscopy. , 1992, Journal of the Optical Society of America. A, Optics and image science.

[21]  Prabhakar R. Gudla,et al.  Segmentation of Whole Cells and Cell Nuclei From 3-D Optical Microscope Images Using Dynamic Programming , 2008, IEEE Transactions on Medical Imaging.

[22]  Paul K Wallace,et al.  Cell Tracking 2007: A Proliferation of Probes and Applications , 2007, Immunological investigations.

[23]  M K Cheezum,et al.  Quantitative comparison of algorithms for tracking single fluorescent particles. , 2001, Biophysical journal.

[24]  Kannappan Palaniappan,et al.  Cell Segmentation Using Coupled Level Sets and Graph-Vertex Coloring , 2006, MICCAI.

[25]  Hanchuan Peng,et al.  Bioimage informatics: a new area of engineering biology , 2008, Bioinform..

[26]  Namrata Vaswani,et al.  Tracking Deforming Objects Using Particle Filtering for Geometric Active Contours , 2007, IEEE Transactions on Pattern Analysis and Machine Intelligence.

[27]  Wiro J. Niessen,et al.  Advanced level-set based multiple-cell segmentation and tracking in time-lapse fluorescence microscopy images , 2008, 2008 5th IEEE International Symposium on Biomedical Imaging: From Nano to Macro.

[28]  Xiaobo Zhou,et al.  Automated segmentation, classification, and tracking of cancer cell nuclei in time-lapse microscopy , 2006, IEEE Transactions on Biomedical Engineering.

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

[30]  Jens Rittscher,et al.  Spatio-temporal cell cycle phase analysis using level sets and fast marching methods , 2009, Medical Image Anal..

[31]  Jouko Lampinen,et al.  Rao-Blackwellized Monte Carlo Data Association for Multiple Target Tracking , 2004 .

[32]  B. C. Carter,et al.  Tracking single particles: a user-friendly quantitative evaluation , 2005, Physical biology.

[33]  Bahram Parvin,et al.  3D Segmentation of Mammospheres for Localization Studies , 2006, ISVC.

[34]  William M. Wells,et al.  Simultaneous truth and performance level estimation (STAPLE): an algorithm for the validation of image segmentation , 2004, IEEE Transactions on Medical Imaging.

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

[36]  Anne E Carpenter,et al.  CellProfiler: free, versatile software for automated biological image analysis. , 2007, BioTechniques.

[37]  R. Waterston,et al.  Automated cell lineage tracing in Caenorhabditis elegans. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[38]  M. Unser,et al.  The colored revolution of bioimaging , 2006, IEEE Signal Processing Magazine.

[39]  Jean-Christophe Olivo-Marin,et al.  On the digital trail of mobile cells , 2006, IEEE Signal Processing Magazine.

[40]  Donna J. Webb,et al.  New dimensions in cell migration , 2003, Nature Cell Biology.

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

[42]  Roger Y Tsien,et al.  Imagining imaging's future. , 2003, Nature reviews. Molecular cell biology.

[43]  S. Deans The Radon Transform and Some of Its Applications , 1983 .

[44]  Douglas B. Kell,et al.  Automatic tracking of biological cells and compartments using particle filters and active contours , 2006 .

[45]  Jean X. Gao,et al.  Multiple Interacting Subcellular Structure Tracking by Sequential Monte Carlo Method , 2007, 2007 IEEE International Conference on Bioinformatics and Biomedicine (BIBM 2007).

[46]  Muriel Golzio,et al.  Tracking in vitro and in vivo siRNA electrotransfer in tumor cells , 2008, Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research.

[47]  Victoria J Allan,et al.  Light Microscopy Techniques for Live Cell Imaging , 2003, Science.

[48]  Douglas G. Altman,et al.  Practical statistics for medical research , 1990 .

[49]  D. Dormann,et al.  Simultaneous quantification of cell motility and protein-membrane-association using active contours. , 2002, Cell motility and the cytoskeleton.

[50]  Richard M Levenson,et al.  Autofluorescence removal, multiplexing, and automated analysis methods for in-vivo fluorescence imaging. , 2005, Journal of biomedical optics.

[51]  Carl-Fredrik Westin,et al.  Fast sub-voxel re-initialization of the distance map for level set methods , 2005, Pattern Recognit. Lett..

[52]  Meng Wang,et al.  Novel cell segmentation and online SVM for cell cycle phase identification in automated microscopy , 2008, Bioinform..

[53]  Rachid Deriche,et al.  Geodesic Active Regions and Level Set Methods for Supervised Texture Segmentation , 2002, International Journal of Computer Vision.

[54]  Erik Meijering,et al.  Chapter 15 – Time‐Lapse Imaging , 2008 .

[55]  Nathalie Harder,et al.  Automated Analysis of the Mitotic Phases of Human Cells in 3D Fluorescence Microscopy Image Sequences , 2006, MICCAI.

[56]  Bing Song Topics in Variational PDE Image Segmentation, Inpainting and Denoising , 2003 .

[57]  Jens Rittscher,et al.  Spatio-temporal cell segmentation and tracking for automated screening , 2008, 2008 5th IEEE International Symposium on Biomedical Imaging: From Nano to Macro.

[58]  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.

[59]  J. Carlin,et al.  Bias, prevalence and kappa. , 1993, Journal of clinical epidemiology.

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

[61]  Ronald Fedkiw,et al.  Level set methods and dynamic implicit surfaces , 2002, Applied mathematical sciences.

[62]  D. Bullard,et al.  Intercellular Adhesion Molecule-1 (ICAM-1) Regulates Endothelial Cell Motility through a Nitric Oxide-dependent Pathway* , 2004, Journal of Biological Chemistry.

[63]  D. Dormann,et al.  Imaging of cell migration , 2006, The EMBO journal.

[64]  Jean-Marc Odobez,et al.  Using particles to track varying numbers of interacting people , 2005, 2005 IEEE Computer Society Conference on Computer Vision and Pattern Recognition (CVPR'05).

[65]  Anne E Carpenter,et al.  CellProfiler: image analysis software for identifying and quantifying cell phenotypes , 2006, Genome Biology.

[66]  Gunilla Borgefors,et al.  Weighted distance transforms for volume images digitized in elongated voxel grids , 2004, Pattern Recognit. Lett..

[67]  Takeo Kanade,et al.  Cell population tracking and lineage construction with spatiotemporal context , 2008, Medical Image Anal..

[68]  Jason R Swedlow,et al.  Time-lapse imaging reveals dynamic relocalization of PP1gamma throughout the mammalian cell cycle. , 2003, Molecular biology of the cell.

[69]  Richard M. Karp Computer Science as a Lens on the Sciences: The Example of Computational Molecular Biology , 2007, 2007 IEEE International Conference on Bioinformatics and Biomedicine (BIBM 2007).