title: Segmentation Method of Time-Lapse Microscopy Images with the Focus on Biocompatibility Assessment.
abstract: Biocompatibility testing of new materials is often performed in vitro by measuring the growth rate of mammalian cancer cells in time-lapse images acquired by phase contrast microscopes. The growth rate is measured by tracking cell coverage, which requires an accurate automatic segmentation method. However, cancer cells have irregular shapes that change over time, the mottled background pattern is partially visible through the cells and the images contain artifacts such as halos. We developed a novel algorithm for cell segmentation that copes with the mentioned challenges. It is based on temporal differences of consecutive images and a combination of thresholding, blurring, and morphological operations. We tested the algorithm on images of four cell types acquired by two different microscopes, evaluated the precision of segmentation against manual segmentation performed by a human operator, and finally provided comparison with other freely available methods. We propose a new, fully automated method for measuring the cell growth rate based on fitting a coverage curve with the Verhulst population model. The algorithm is fast and shows accuracy comparable with manual segmentation. Most notably it can correctly separate live from dead cells.
authors.affiliation: 1Institute of Complex Systems FFPW, CENAKVA,University of South Bohemia,Zámek 136,CZ-373 33 Nové Hrady,Czech Republic.
authors.forename: Jindřich
authors.initials: J
authors.lastname: Soukup
authors.affiliation: 1Institute of Complex Systems FFPW, CENAKVA,University of South Bohemia,Zámek 136,CZ-373 33 Nové Hrady,Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Císař
authors.affiliation: 3Department of Image Processing,Institute of Information Theory and Automation of the ASCR,Pod vodárenskou věží 4,CZ-182 08 Prague 8,Czech Republic.
authors.forename: Filip
authors.initials: F
authors.lastname: Šroubek
mesh_terms: D000465:Algorithms; D000818:Animals; D016477:Artifacts; D003584:Cytological Techniques / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D008853:Microscopy; D010363:Pattern Recognition, Automated; D059008:Time-Lapse Imaging
keywords: biocompatibility assessment; cytotoxicity testing; phase contrast microscopy; segmentation; time-lapse