Journal article
High-resolution 3D imaging of fixed and cleared organoids
JF Dekkers, M Alieva, LM Wellens, HCR Ariese, PR Jamieson, AM Vonk, GD Amatngalim, H Hu, KC Oost, HJG Snippert, JM Beekman, EJ Wehrens, JE Visvader, H Clevers, AC Rios
Nature Protocols | NATURE PUBLISHING GROUP | Published : 2019
Abstract
In vitro 3D organoid systems have revolutionized the modeling of organ development and diseases in a dish. Fluorescence microscopy has contributed to the characterization of the cellular composition of organoids and demonstrated organoids’ phenotypic resemblance to their original tissues. Here, we provide a detailed protocol for performing high-resolution 3D imaging of entire organoids harboring fluorescence reporters and upon immunolabeling. This method is applicable to a wide range of organoids of differing origins and of various sizes and shapes. We have successfully used it on human airway, colon, kidney, liver and breast tumor organoids, as well as on mouse mammary gland organoids. It i..
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Funding Acknowledgements
We are very grateful for the technical support from the Princess Maxima Center for Pediatric Oncology and to the Hubrecht Institute and Zeiss for imaging support and collaborations. All the imaging was performed at the Princess Maxima imaging center. This work was financially supported by the Princess Maxima Center for Pediatric Oncology. J.F.D. was supported by a Marie Curie Global Fellowship and a VENI grant from the Netherlands Organisation for Scientific Research (NWO). J.E.V. was supported by the Australian National Health and Medical Research Council (NHMRC).