@article{7fdc220f47304870bacc43cdb68846ab,
title = "Single-Cell Resolution Three-Dimensional Imaging of Intact Organoids",
abstract = "Organoid technology, in vitro 3D culturing of miniature tissue, has opened a new experimental window for cellular processes that govern organ development and function as well as disease. Fluorescence microscopy has played a major role in characterizing their cellular composition in detail and demonstrating their similarity to the tissue they originate from. In this article, we present a comprehensive protocol for high-resolution 3D imaging of whole organoids upon immunofluorescent labeling. This method is widely applicable for imaging of organoids differing in origin, size and shape. Thus far we have applied the method to airway, colon, kidney, and liver organoids derived from healthy human tissue, as well as human breast tumor organoids and mouse mammary gland organoids. We use an optical clearing agent, FUnGI, which enables the acquisition of whole 3D organoids with the opportunity for single-cell quantification of markers. This three-day protocol from organoid harvesting to image analysis is optimized for 3D imaging using confocal microscopy.",
keywords = "Animals, Humans, Imaging, Three-Dimensional/methods, Mice, Organoids/diagnostic imaging, 3D imaging, Optical clearing, Confocal microscopy, Organoid, Biology, Single-cell resolution, Immunolabelling, Issue 160",
author = "\{van Ineveld\}, \{Ravian L\} and Ariese, \{Hendrikus C R\} and Wehrens, \{Ellen J\} and Dekkers, \{Johanna F\} and Rios, \{Anne C\}",
note = "Funding Information: We are very grateful for the technical support from the Princess M{\'a}xima Center for Pediatric Oncology and to the Hubrecht Institute and Zeiss for imaging support and collaborations. All the imaging was performed at the Princess M{\'a}xima imaging center. This work was financially supported by the Princess M{\'a}xima Center for Pediatric Oncology. JFD was supported by a Marie Curie Global Fellowship and a VENI grant from the Netherlands Organisation for Scientific Research (NWO). ACR was supported by a European Council (ERC) starting grant. Funding Information: We are very grateful for the technical support from the Princess M?xima Center for Pediatric Oncology and to the Hubrecht Institute and Zeiss for imaging support and collaborations. All the imaging was performed at the Princess M?xima imaging center. This work was financially supported by the Princess M?xima Center for Pediatric Oncology. JFD was supported by a Marie Curie Global Fellowship and a VENI grant from the Netherlands Organisation for Scientific Research (NWO). ACR was supported by a European Council (ERC) starting grant. Publisher Copyright: {\textcopyright} 2020 Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License.",
year = "2020",
month = jun,
day = "5",
doi = "10.3791/60709",
language = "English",
volume = "2020",
pages = "1--8",
journal = "Journal of visualized experiments : JoVE",
issn = "1940-087X",
publisher = "MYJoVE Corporation",
number = "160",
}