title: Novel fixedz-direction (FiZD) kidney primordia and an organoid culture system for time-lapse confocal imaging.
abstract: Tissue, organ and organoid cultures provide suitable models for developmental studies, but our understanding of how the organs are assembled at the single-cell level still remains unclear. We describe here a novel fixedz-direction (FiZD) culture setup that permits high-resolution confocal imaging of organoids and embryonic tissues. In a FiZD culture a permeable membrane compresses the tissues onto a glass coverslip and the spacers adjust the thickness, enabling the tissue to grow for up to 12 days. Thus, the kidney rudiment and the organoids can adjust to the limitedz-directional space and yet advance the process of kidney morphogenesis, enabling long-term time-lapse and high-resolution confocal imaging. As the data quality achieved was sufficient for computer-assisted cell segmentation and analysis, the method can be used for studying morphogenesisex vivoat the level of the single constituent cells of a complex mammalian organogenesis model system.
authors.affiliation: Faculty of Biochemistry and Molecular Medicine, University of Oulu, 90220 Oulu, Finland.|Laboratory of Developmental Biology, Biocenter Oulu and InfoTech, 90220 Oulu, Finland.|Department of Medical Biochemistry and Molecular Medicine, Oulu Center for Cell Matrix Research, 90220 Oulu, Finland.
authors.forename: Ulla
authors.identifier: 0000-0002-9410-6548
authors.initials: U
authors.lastname: Saarela
authors.affiliation: Laboratory of Developmental Biology, Biocenter Oulu and InfoTech, 90220 Oulu, Finland.|Center for Machine Vision Research, Department of Computer Science and Engineering, University of Oulu, 90014 Oulu, Finland.
authors.forename: Saad Ullah
authors.initials: SU
authors.lastname: Akram
authors.affiliation: Sorbonne Universités, UPMC Univ Paris 06, IBPS - UMR7622 Developmental Biology, Paris F-75005, France.|Institut de Biologie Paris-Seine (IBPS) - CNRS UMR7622 Developmental Biology, F-75005 Paris, France.
authors.forename: Audrey
authors.identifier: 0000-0001-9716-755X
authors.initials: A
authors.lastname: Desgrange
authors.affiliation: Faculty of Biochemistry and Molecular Medicine, University of Oulu, 90220 Oulu, Finland.|Laboratory of Developmental Biology, Biocenter Oulu and InfoTech, 90220 Oulu, Finland.|Department of Medical Biochemistry and Molecular Medicine, Oulu Center for Cell Matrix Research, 90220 Oulu, Finland.
authors.forename: Aleksandra
authors.identifier: 0000-0002-2399-4410
authors.initials: A
authors.lastname: Rak-Raszewska
authors.affiliation: Faculty of Biochemistry and Molecular Medicine, University of Oulu, 90220 Oulu, Finland.|Laboratory of Developmental Biology, Biocenter Oulu and InfoTech, 90220 Oulu, Finland.|Department of Medical Biochemistry and Molecular Medicine, Oulu Center for Cell Matrix Research, 90220 Oulu, Finland.
authors.forename: Jingdong
authors.initials: J
authors.lastname: Shan
authors.affiliation: Sorbonne Universités, UPMC Univ Paris 06, IBPS - UMR7622 Developmental Biology, Paris F-75005, France.|Institut de Biologie Paris-Seine (IBPS) - CNRS UMR7622 Developmental Biology, F-75005 Paris, France.
authors.forename: Silvia
authors.initials: S
authors.lastname: Cereghini
authors.affiliation: Biocenter Oulu, University of Oulu, 90220 Oulu, Finland.
authors.forename: Veli-Pekka
authors.identifier: 0000-0003-3050-4579
authors.initials: VP
authors.lastname: Ronkainen
authors.affiliation: Center for Machine Vision Research, Department of Computer Science and Engineering, University of Oulu, 90014 Oulu, Finland.
authors.forename: Janne
authors.initials: J
authors.lastname: Heikkilä
authors.affiliation: Faculty of Biochemistry and Molecular Medicine, University of Oulu, 90220 Oulu, Finland ilya.skovorodkin@oulu.fi seppo.vainio@oulu.fi.|Laboratory of Developmental Biology, Biocenter Oulu and InfoTech, 90220 Oulu, Finland.|Department of Medical Biochemistry and Molecular Medicine, Oulu Center for Cell Matrix Research, 90220 Oulu, Finland.
authors.forename: Ilya
authors.identifier: 0000-0003-1182-1130
authors.initials: I
authors.lastname: Skovorodkin
authors.affiliation: Faculty of Biochemistry and Molecular Medicine, University of Oulu, 90220 Oulu, Finland ilya.skovorodkin@oulu.fi seppo.vainio@oulu.fi.|Laboratory of Developmental Biology, Biocenter Oulu and InfoTech, 90220 Oulu, Finland.|Department of Medical Biochemistry and Molecular Medicine, Oulu Center for Cell Matrix Research, 90220 Oulu, Finland.
authors.forename: Seppo J
authors.identifier: 0000-0001-9319-3566
authors.initials: SJ
authors.lastname: Vainio
mesh_terms: D000818:Animals; D021621:Imaging, Three-Dimensional; D007668:Kidney / Q000196:embryology*; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods*; D009024:Morphogenesis; D009940:Organoids / Q000196:embryology*; D059008:Time-Lapse Imaging / Q000379:methods*; D046509:Tissue Culture Techniques / Q000379:methods*
keywords: Kidney; Organ culture; Organoid, Imaging; Time-lapse