IRL 1 article published in JoVE Biology A High-Throughput Platform for Culture and 3D Imaging of Organoids Gianluca Grenci1,2, Florian Dilasser1, Saburnisha Binte Mohamad Raffi1, Marion Marchand1, Mona Suryana1, Geetika Sahni1, Virgile Viasnoff1,3,4, Anne Beghin1,5 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 This paper presents a fabrication protocol for a new type of culture substrate with hundreds of microcontainers per mm2, in which organoids can be cultured and observed using high-resolution microscopy. The cell seeding and immunostaining protocols are also detailed.