Novel Fixed z-direction (FiZD) Kidney Primordia and an Organoid Culture System for Time-Lapse Confocal Imaging

Development. 2017 Mar 15;144(6):1113-1117. doi: 10.1242/dev.142950. Epub 2017 Feb 20.

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 fixed z-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 limited z-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 morphogenesis ex vivo at the level of the single constituent cells of a complex mammalian organogenesis model system.

Keywords: Kidney; Organ culture; Organoid, Imaging; Time-lapse.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Imaging, Three-Dimensional
  • Kidney / embryology*
  • Mice
  • Microscopy, Confocal / methods*
  • Morphogenesis
  • Organoids / embryology*
  • Time-Lapse Imaging / methods*
  • Tissue Culture Techniques / methods*