3D fluorescence staining and confocal imaging of low amount of intestinal organoids (enteroids): Protocol accessible to all.
The emerging field of 3D organ modeling encounters several imaging issues in particular related to antigen retrieval and sample loss during staining processes. Due to their compact shape, several antibodies fail to penetrate intact organoids or spheroids. Histology of organoids can be approached by...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2025-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0315922 |
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| author | Ami Gloria Toulehohoun Caroline Bouzin Aurélie Daumerie Luca Maccioni Peter Stärkel |
| author_facet | Ami Gloria Toulehohoun Caroline Bouzin Aurélie Daumerie Luca Maccioni Peter Stärkel |
| author_sort | Ami Gloria Toulehohoun |
| collection | DOAJ |
| description | The emerging field of 3D organ modeling encounters several imaging issues in particular related to antigen retrieval and sample loss during staining processes. Due to their compact shape, several antibodies fail to penetrate intact organoids or spheroids. Histology of organoids can be approached by paraffin inclusion and sectioning at 5 μm as performed for biopsies. However, to fully understand organoid behavior, including cellular organization, extracellular matrix structure, and their response to treatments, 3D imaging is essential. Here we propose an easy workflow allowing (1) immunostaining with a HIER step, (2) preservation of the intact shape of the organoids, (3) sample immobilization in a focal plane reachable for high resolution/short working distance lenses, and (4) minimizing the risk of loss of precious material. |
| format | Article |
| id | doaj-art-0fa17b86b6534c20b5d3fed1df16b554 |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-0fa17b86b6534c20b5d3fed1df16b5542025-08-20T02:12:46ZengPublic Library of Science (PLoS)PLoS ONE1932-62032025-01-01201e031592210.1371/journal.pone.03159223D fluorescence staining and confocal imaging of low amount of intestinal organoids (enteroids): Protocol accessible to all.Ami Gloria ToulehohounCaroline BouzinAurélie DaumerieLuca MaccioniPeter StärkelThe emerging field of 3D organ modeling encounters several imaging issues in particular related to antigen retrieval and sample loss during staining processes. Due to their compact shape, several antibodies fail to penetrate intact organoids or spheroids. Histology of organoids can be approached by paraffin inclusion and sectioning at 5 μm as performed for biopsies. However, to fully understand organoid behavior, including cellular organization, extracellular matrix structure, and their response to treatments, 3D imaging is essential. Here we propose an easy workflow allowing (1) immunostaining with a HIER step, (2) preservation of the intact shape of the organoids, (3) sample immobilization in a focal plane reachable for high resolution/short working distance lenses, and (4) minimizing the risk of loss of precious material.https://doi.org/10.1371/journal.pone.0315922 |
| spellingShingle | Ami Gloria Toulehohoun Caroline Bouzin Aurélie Daumerie Luca Maccioni Peter Stärkel 3D fluorescence staining and confocal imaging of low amount of intestinal organoids (enteroids): Protocol accessible to all. PLoS ONE |
| title | 3D fluorescence staining and confocal imaging of low amount of intestinal organoids (enteroids): Protocol accessible to all. |
| title_full | 3D fluorescence staining and confocal imaging of low amount of intestinal organoids (enteroids): Protocol accessible to all. |
| title_fullStr | 3D fluorescence staining and confocal imaging of low amount of intestinal organoids (enteroids): Protocol accessible to all. |
| title_full_unstemmed | 3D fluorescence staining and confocal imaging of low amount of intestinal organoids (enteroids): Protocol accessible to all. |
| title_short | 3D fluorescence staining and confocal imaging of low amount of intestinal organoids (enteroids): Protocol accessible to all. |
| title_sort | 3d fluorescence staining and confocal imaging of low amount of intestinal organoids enteroids protocol accessible to all |
| url | https://doi.org/10.1371/journal.pone.0315922 |
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