Automated high speed 3D imaging of organoid cultures with efficient multi-scale phenotypic quantification

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Beghin, Anne | Grenci, Gianluca | Sahni, Geetika | Guo, Su | Rajendiran, Harini | Delaire, Tom | Mohamad Raffi, Saburnisha Binte | Blanc, Damien | de Mets, Richard | Ong, Hui Ting | Galindo, Xareni | Monet, Anais | Acharya, Vidhyalakshmi | Racine, Victor | Levet, Florian | Galland, Remi | Sibarita, Jean-Baptiste | Viasnoff, Virgile

Edité par CCSD ; Nature Publishing Group -

International audience. Current imaging approaches limit the ability to perform multiscale characterization of 3D organotypic cultures (organoids) in large numbers. Here, we present an automated multiscale 3D imaging platform synergizing high-density organoid cultures with 3D live single objective light-sheet imaging. It is composed of disposable microfabricated organoid culture chips embedding optical components and a custom laser beam steering unit coupled to a commercial inverted microscope. It streamlines organoid culture and high content 3D imaging on a single user-friendly instrument with minimal manipulations and unprecedented throughput of 300 organoids per hour in 3D. Collecting large number of 3D stacks allowed training deep learning-based algorithms to quantify the organoids morphogenetic organizations at multi-scales, ranging from the sub-cellular scale to the whole organoid level. We validated the versatility and robustness of our approach on intestine, hepatic, neuroectoderm organoids and oncospheres.

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