Novel fixed z-direction (FiZD) kidney primordia and an organoid culture system for time-lapse confocal imaging
Saarela, Ulla; Akram, Saad Ullah; Desgrange, Audrey; Rak-Raszewska, Aleksandra; Shan, Jingdong; Cereghini, Silvia; Ronkainen, Veli-Pekka; Heikkilä, Janne; Skovorodkin, Ilya; Vainio, Seppo J. (2017-03-01)
Ulla Saarela, Saad Ullah Akram, Audrey Desgrange, Aleksandra Rak-Raszewska, Jingdong Shan, Silvia Cereghini, Veli-Pekka Ronkainen, Janne Heikkilä, Ilya Skovorodkin, Seppo J. Vainio. Novel fixed z-direction (FiZD) kidney primordia and an organoid culture system for time-lapse confocal imaging. Development 2017 144: 1113-1117; doi: 10.1242/dev.142950
© 2017. Published by The Company of Biologists Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
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.
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