Cerebral Organoid Arrays for Batch Phenotypic Analysis in Sections and Three Dimensions
Organoids can recapitulate human-specific phenotypes and functions in vivo and have great potential for research in development, disease modeling, and drug screening. Due to the inherent variability among organoids, experiments often require a large sample size. Embedding, staining, and imaging each...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-09-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/18/13903 |
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| author | Juan Chen Haihua Ma Zhiyu Deng Qingming Luo Hui Gong Ben Long Xiangning Li |
| author_facet | Juan Chen Haihua Ma Zhiyu Deng Qingming Luo Hui Gong Ben Long Xiangning Li |
| author_sort | Juan Chen |
| collection | DOAJ |
| description | Organoids can recapitulate human-specific phenotypes and functions in vivo and have great potential for research in development, disease modeling, and drug screening. Due to the inherent variability among organoids, experiments often require a large sample size. Embedding, staining, and imaging each organoid individually require a lot of reagents and time. Hence, there is an urgent need for fast and efficient methods for analyzing the phenotypic changes in organoids in batches. Here, we provide a comprehensive strategy for array embedding, staining, and imaging of cerebral organoids in both agarose sections and in 3D to analyze the spatial distribution of biomarkers in organoids in situ. We constructed several disease models, particularly an aging model, as examples to demonstrate our strategy for the investigation of the phenotypic analysis of organoids. We fabricated an array mold to produce agarose support with microwells, which hold organoids in place for live/dead imaging. We performed staining and imaging of sectioned organoids embedded in agarose and 3D imaging to examine phenotypic changes in organoids using fluorescence micro-optical sectioning tomography (fMOST) and whole-mount immunostaining. Parallel studies of organoids in arrays using the same staining and imaging parameters enabled easy and reliable comparison among different groups. We were able to track all the data points obtained from every organoid in an embedded array. This strategy could help us study the phenotypic changes in organoids in disease models and drug screening. |
| first_indexed | 2024-03-10T22:39:56Z |
| format | Article |
| id | doaj.art-0ff24311ae0045df987a6bf44aee81b0 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T22:39:56Z |
| publishDate | 2023-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-0ff24311ae0045df987a6bf44aee81b02023-11-19T11:04:52ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-09-0124181390310.3390/ijms241813903Cerebral Organoid Arrays for Batch Phenotypic Analysis in Sections and Three DimensionsJuan Chen0Haihua Ma1Zhiyu Deng2Qingming Luo3Hui Gong4Ben Long5Xiangning Li6Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan 430074, ChinaBritton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan 430074, ChinaKey Laboratory of Biomedical Engineering of Hainan Province, School of Biomedical Engineering, Hainan University, Haikou 570228, ChinaKey Laboratory of Biomedical Engineering of Hainan Province, School of Biomedical Engineering, Hainan University, Haikou 570228, ChinaBritton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan 430074, ChinaKey Laboratory of Biomedical Engineering of Hainan Province, School of Biomedical Engineering, Hainan University, Haikou 570228, ChinaKey Laboratory of Biomedical Engineering of Hainan Province, School of Biomedical Engineering, Hainan University, Haikou 570228, ChinaOrganoids can recapitulate human-specific phenotypes and functions in vivo and have great potential for research in development, disease modeling, and drug screening. Due to the inherent variability among organoids, experiments often require a large sample size. Embedding, staining, and imaging each organoid individually require a lot of reagents and time. Hence, there is an urgent need for fast and efficient methods for analyzing the phenotypic changes in organoids in batches. Here, we provide a comprehensive strategy for array embedding, staining, and imaging of cerebral organoids in both agarose sections and in 3D to analyze the spatial distribution of biomarkers in organoids in situ. We constructed several disease models, particularly an aging model, as examples to demonstrate our strategy for the investigation of the phenotypic analysis of organoids. We fabricated an array mold to produce agarose support with microwells, which hold organoids in place for live/dead imaging. We performed staining and imaging of sectioned organoids embedded in agarose and 3D imaging to examine phenotypic changes in organoids using fluorescence micro-optical sectioning tomography (fMOST) and whole-mount immunostaining. Parallel studies of organoids in arrays using the same staining and imaging parameters enabled easy and reliable comparison among different groups. We were able to track all the data points obtained from every organoid in an embedded array. This strategy could help us study the phenotypic changes in organoids in disease models and drug screening.https://www.mdpi.com/1422-0067/24/18/13903cerebral organoidsarray imagingsingle-cell phenotypic analysisaging |
| spellingShingle | Juan Chen Haihua Ma Zhiyu Deng Qingming Luo Hui Gong Ben Long Xiangning Li Cerebral Organoid Arrays for Batch Phenotypic Analysis in Sections and Three Dimensions International Journal of Molecular Sciences cerebral organoids array imaging single-cell phenotypic analysis aging |
| title | Cerebral Organoid Arrays for Batch Phenotypic Analysis in Sections and Three Dimensions |
| title_full | Cerebral Organoid Arrays for Batch Phenotypic Analysis in Sections and Three Dimensions |
| title_fullStr | Cerebral Organoid Arrays for Batch Phenotypic Analysis in Sections and Three Dimensions |
| title_full_unstemmed | Cerebral Organoid Arrays for Batch Phenotypic Analysis in Sections and Three Dimensions |
| title_short | Cerebral Organoid Arrays for Batch Phenotypic Analysis in Sections and Three Dimensions |
| title_sort | cerebral organoid arrays for batch phenotypic analysis in sections and three dimensions |
| topic | cerebral organoids array imaging single-cell phenotypic analysis aging |
| url | https://www.mdpi.com/1422-0067/24/18/13903 |
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