title: Construction of three quadruple-fluorescent MDA435 cell lines that enable monitoring of the whole chromosome segregation process in the living state.
abstract: Mitotic events from prophase to telophase are defined by morphology or movement of chromatin, nuclear envelope, centrosomes and spindles. Live-cell imaging is useful for characterizing the whole chromosome segregation process in the living state. In this study, we constructed three quadruple-fluorescent MDA435 cell lines in which chromatin, kinetochores, nuclear envelope and either inner centromere, microtubules or centrosomes/spindles were differentially visualized with cyan, green, orange and red fluorescent proteins (ECFP, EGFP, mKO and DsRed). Each mitotic stage of the individual cells could be identified by capturing live-cell images without the requirement of fixing or staining steps. In addition, we obtained four-color time-lapse images of one cell line, MDA-Auro/imp/H3/AF, from prophase to metaphase and from early anaphase to telophase. These quadruple-fluorescent cell lines will be useful for precisely analyzing the mitotic events from prophase through to telophase in single cells in the future.
authors.affiliation: Live Cell Imaging Institute, Osaka Prefecture University, Sakai, Osaka 599-8531, Japan. sugimoto@bioinfo.osakafu-u.ac.jp
authors.forename: Kenji
authors.initials: K
authors.lastname: Sugimoto
authors.forename: Kaori
authors.initials: K
authors.lastname: Senda-Murata
authors.forename: Shigenori
authors.initials: S
authors.lastname: Oka
mesh_terms: D015415:Biomarkers / Q000032:analysis; D002460:Cell Line; D002843:Chromatin / Q000648:ultrastructure; D020090:Chromosome Segregation; D005453:Fluorescence; D006801:Humans; D018386:Kinetochores / Q000648:ultrastructure; D008938:Mitosis; D008957:Models, Genetic; D009685:Nuclear Envelope / Q000648:ultrastructure; D008941:Spindle Apparatus / Q000648:ultrastructure
chemical_list: D015415:Biomarkers; D002843:Chromatin