Single-Cell Resolution Three-Dimensional Imaging of Intact Organoids
| Autor: | Ravian L. van Ineveld, Hendrikus C R Ariese, Ellen J. Wehrens, Anne C. Rios, Johanna F. Dekkers |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
General Chemical Engineering Cell Mouse Mammary Gland Biology General Biochemistry Genetics and Molecular Biology law.invention 03 medical and health sciences Mice 0302 clinical medicine Imaging Three-Dimensional Confocal microscopy law Optical clearing medicine Fluorescence microscope Organoid Animals Humans General Immunology and Microbiology General Neuroscience Resolution (electron density) Cell biology Organoids 030104 developmental biology medicine.anatomical_structure Three dimensional imaging 030217 neurology & neurosurgery |
| Zdroj: | Journal of visualized experiments : JoVE. (160) |
| ISSN: | 1940-087X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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