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


Current imaging approaches limit the ability to perform multi-scale characterization of three-dimensional (3D) organotypic cultures (organoids) in large numbers. Here, we present an automated multi-scale 3D imaging platform synergizing high-density organoid cultures with rapid and live 3D single-objective light-sheet imaging. It is composed of disposable microfabricated organoid culture chips, termed JeWells, with embedded optical components and a laser beam-steering unit coupled to a commercial inverted microscope. It permits streamlining organoid culture and high-content 3D imaging on a single user-friendly instrument with minimal manipulations and a throughput of 300 organoids per hour. We demonstrate that the large number of 3D stacks that can be collected via our platform allows training deep learning-based algorithms to quantify morphogenetic organizations of organoids at multi-scales, ranging from the subcellular scale to the whole organoid level. We validated the versatility and robustness of our approach on intestine, hepatic, neuroectoderm organoids and oncospheres. © 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.

Authors Beghin A, Grenci G, Sahni G, Guo S, Rajendiran H, Delaire T, Mohamad Raffi SB, Blanc D, de Mets R, Ong HT, Galindo X, Monet A, Acharya V, Racine V, Levet F, Galland R, Sibarita JB, Viasnoff V
Journal Nature methods
Publication Date 2022 Jul;19(7):881-892
PubMed 35697835
DOI 10.1038/s41592-022-01508-0

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