title: Accurate Morphology Preserving Segmentation of Overlapping Cells based on Active Contours.
abstract: The identification of fluorescently stained cell nuclei is the basis of cell detection, segmentation, and feature extraction in high content microscopy experiments. The nuclear morphology of single cells is also one of the essential indicators of phenotypic variation. However, the cells used in experiments can lose their contact inhibition, and can therefore pile up on top of each other, making the detection of single cells extremely challenging using current segmentation methods. The model we present here can detect cell nuclei and their morphology even in high-confluency cell cultures with many overlapping cell nuclei. We combine the "gas of near circles" active contour model, which favors circular shapes but allows slight variations around them, with a new data model. This captures a common property of many microscopic imaging techniques: the intensities from superposed nuclei are additive, so that two overlapping nuclei, for example, have a total intensity that is approximately double the intensity of a single nucleus. We demonstrate the power of our method on microscopic images of cells, comparing the results with those obtained from a widely used approach, and with manual image segmentations by experts.
authors: Synthetic and System Biology Unit, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary.
authors: Csaba
authors: C
authors: Molnar
authors: Department of Mathematical Sciences, Durham University, Durham, UK.
authors: Ian H
authors: IH
authors: Jermyn
authors: Department of Mathematics and Informatics, J. Selye University, Komarno, Slovakia.
authors: Zoltan
authors: Z
authors: Kato
authors: Institute for Molecular Medicine Finland, University of Helsinki, Helsinki, Finland.
authors: Vesa
authors: V
authors: Rahkama
authors: Institute for Molecular Medicine Finland, University of Helsinki, Helsinki, Finland.
authors: Päivi
authors: P
authors: Östling
authors: Institute for Molecular Medicine Finland, University of Helsinki, Helsinki, Finland.
authors: Piia
authors: P
authors: Mikkonen
authors: Institute for Molecular Medicine Finland, University of Helsinki, Helsinki, Finland.
authors: Vilja
authors: V
authors: Pietiäinen
authors: Synthetic and System Biology Unit, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary.|Institute for Molecular Medicine Finland, University of Helsinki, Helsinki, Finland.
authors: Peter
authors: P
authors: Horvath
mesh_terms: D000465:Algorithms; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism*; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D008954:Models, Biological; D015388:Organelles / Q000378:metabolism*; D015203:Reproducibility of Results; D059010:Single-Cell Analysis / Q000379:methods*