title: Mathematical imaging methods for mitosis analysis in live-cell phase contrast microscopy.
abstract: In this paper we propose a workflow to detect and track mitotic cells in time-lapse microscopy image sequences. In order to avoid the requirement for cell lines expressing fluorescent markers and the associated phototoxicity, phase contrast microscopy is often preferred over fluorescence microscopy in live-cell imaging. However, common specific image characteristics complicate image processing and impede use of standard methods. Nevertheless, automated analysis is desirable due to manual analysis being subjective, biased and extremely time-consuming for large data sets. Here, we present the following workflow based on mathematical imaging methods. In the first step, mitosis detection is performed by means of the circular Hough transform. The obtained circular contour subsequently serves as an initialisation for the tracking algorithm based on variational methods. It is sub-divided into two parts: in order to determine the beginning of the whole mitosis cycle, a backwards tracking procedure is performed. After that, the cell is tracked forwards in time until the end of mitosis. As a result, the average of mitosis duration and ratios of different cell fates (cell death, no division, division into two or more daughter cells) can be measured and statistics on cell morphologies can be obtained. All of the tools are featured in the user-friendly MATLAB®Graphical User Interface MitosisAnalyser.
authors.affiliation: University of Cambridge, Department of Applied Mathematics and Theoretical Physics, Centre for Mathematical Sciences, Wilberforce Road, Cambridge CB3 0WA, United Kingdom. Electronic address: jg704@cam.ac.uk.
authors.forename: Joana Sarah
authors.initials: JS
authors.lastname: Grah
authors.affiliation: University of Cambridge, Cancer Research UK Cambridge Institute, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, United Kingdom.
authors.forename: Jennifer Alison
authors.initials: JA
authors.lastname: Harrington
authors.affiliation: University of Cambridge, Cancer Research UK Cambridge Institute, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, United Kingdom.
authors.forename: Siang Boon
authors.initials: SB
authors.lastname: Koh
authors.affiliation: University of Cambridge, Cancer Research UK Cambridge Institute, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, United Kingdom.
authors.forename: Jeremy Andrew
authors.initials: JA
authors.lastname: Pike
authors.affiliation: University of Cambridge, Cancer Research UK Cambridge Institute, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, United Kingdom.
authors.forename: Alexander
authors.initials: A
authors.lastname: Schreiner
authors.affiliation: Westfälische Wilhelms-Universität Münster, Institute for Computational and Applied Mathematics, Einsteinstrasse 62, 48149 Münster, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Burger
authors.affiliation: University of Cambridge, Department of Applied Mathematics and Theoretical Physics, Centre for Mathematical Sciences, Wilberforce Road, Cambridge CB3 0WA, United Kingdom.
authors.forename: Carola-Bibiane
authors.initials: CB
authors.lastname: Schönlieb
authors.affiliation: University of Cambridge, Cancer Research UK Cambridge Institute, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, United Kingdom.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Reichelt
mesh_terms: D000465:Algorithms; D045744:Cell Line, Tumor; D058948:Cell Tracking / Q000379:methods* / Q000706:statistics & numerical data; D004847:Epithelial Cells / Q000648:ultrastructure*; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods* / Q000706:statistics & numerical data; D050417:Insulin-Secreting Cells / Q000648:ultrastructure*; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D008858:Microscopy, Phase-Contrast / Q000295:instrumentation / Q000379:methods*; D008938:Mitosis; D059008:Time-Lapse Imaging / Q000295:instrumentation / Q000379:methods; D057188:Workflow
keywords: Cell tracking; Circular Hough transform; Level-set methods; Mitosis analysis; Phase contrast microscopy; Variational methods