10.17867/10000110a
Ex vivo live cell tracking in kidney organoids using light sheet fluorescence microscopy
Marie Held
0000-0003-0118-5898
Institute of Integrative Biology, University of Liverpool
University of Dundee
2017
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Renal re-aggregation, organoid, light sheet fluorescence microscopy, nephron, differentiation, glomeruli, podocytes, live imaging, cell tracking
Ilaria Santeramo
Institute of Translational Medicine, University of Liverpool
Bettina Wilm
0000-0002-9245-993X
Institute of Translational Medicine, University of Liverpool
Patricia Murray
0000-0002-9245-993X
Institute of Translational Medicine, University of Liverpool
Raphaël Lévy
0000-0001-5728-0531
Institute of Integrative Biology, University of Liverpool
2017-11-08
We have adapted the mouse kidney rudiment assay to generate renal organoids. To demonstrate that the organoid culture method of re-aggregated kidney rudiments in PDMS discs resulted in 3D kidney organoids with organotypic structures, we stained fixed six day old organoids for various developmental markers and imaged them with a light sheet fluorescent microscope. We have stained for the following markers: Megalin, Laminin, Cytokeratin, Pax2, Six2, Wt1, Synaptopodin and Nephrin and have also used the lectin stain PNA (peanut agglutinin). The spheroids were usually imaged as z stacks from five angles.
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