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Abstract

Biology

A High-Throughput Platform for Culture and 3D Imaging of Organoids

Published: October 14th, 2022

DOI:

10.3791/64405

1Mechanobiology Institute (MBI), National University of Singapore, 2Biomedical Engineering Department, National University of Singapore, 3Department of Biological Sciences, National University of Singapore, 4IRL 3639 CNRS, 5Department of Microbiology & Immunology, Immunology Translational Research Program, Yong Loo Lin School of Medicine, National University of Singapore

Abstract

The characterization of a large number of three-dimensional (3D) organotypic cultures (organoids) at different resolution scales is currently limited by standard imaging approaches. This protocol describes a way to prepare microfabricated organoid culture chips, which enable multiscale, 3D live imaging on a user-friendly instrument requiring minimal manipulations and capable of up to 300 organoids/h imaging throughput. These culture chips are compatible with both air and immersion objectives (air, water, oil, and silicone) and a wide range of common microscopes (e.g., spinning disk, point scanner confocal, wide field, and brightfield). Moreover, they can be used with light-sheet modalities such as the single-objective, single-plane illumination microscopy (SPIM) technology (soSPIM).

The protocol described here gives detailed steps for the preparation of the microfabricated culture chips and the culture and staining of organoids. Only a short length of time is required to become familiar with, and consumables and equipment can be easily found in normal biolabs. Here, the 3D imaging capabilities will be demonstrated only with commercial standard microscopes (e.g., spinning disk for 3D reconstruction and wide field microscopy for routine monitoring).

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Keywords High throughput

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