Protocol for immunofluorescent staining, high-resolution imaging, and spatial projection of Schlemm’s canal in mouse whole-mount corneal limbus tissue
Summary: Schlemm’s canal (SC) is an endothelium-lined conduit with uniquely combined features of blood vessels and lymphatic vessels required for aqueous humor drainage from the eye. Here, we present a protocol for immunofluorescent staining of blood and lymphatic endothelial markers in mouse whole-...
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Elsevier
2025-09-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725003120 |
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| author | Weifa Wang Peiyao Peng Wenfo Hu Xuyang Liu Liming Yang Rong Ju Feng Zhang |
| author_facet | Weifa Wang Peiyao Peng Wenfo Hu Xuyang Liu Liming Yang Rong Ju Feng Zhang |
| author_sort | Weifa Wang |
| collection | DOAJ |
| description | Summary: Schlemm’s canal (SC) is an endothelium-lined conduit with uniquely combined features of blood vessels and lymphatic vessels required for aqueous humor drainage from the eye. Here, we present a protocol for immunofluorescent staining of blood and lymphatic endothelial markers in mouse whole-mount anterior eye cup at different postnatal ages. We describe steps for tissue harvesting, fixation, antibody incubation, mounting, and high-resolution confocal microscopy. This protocol facilitates the understanding of SC formation and functionality at the cellular level.For complete details on the use and execution of this protocol, please refer to Gu et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-695a16f8b8f34d65adacbb4df4b79dd6 |
| institution | OA Journals |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-695a16f8b8f34d65adacbb4df4b79dd62025-08-20T02:10:03ZengElsevierSTAR Protocols2666-16672025-09-016310390610.1016/j.xpro.2025.103906Protocol for immunofluorescent staining, high-resolution imaging, and spatial projection of Schlemm’s canal in mouse whole-mount corneal limbus tissueWeifa Wang0Peiyao Peng1Wenfo Hu2Xuyang Liu3Liming Yang4Rong Ju5Feng Zhang6State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, ChinaState Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, ChinaSchool of Basic Medicine, Haerbin Medical University, Haerbin 150081, ChinaState Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, ChinaSchool of Basic Medicine, Haerbin Medical University, Haerbin 150081, ChinaState Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, ChinaState Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China; Corresponding authorSummary: Schlemm’s canal (SC) is an endothelium-lined conduit with uniquely combined features of blood vessels and lymphatic vessels required for aqueous humor drainage from the eye. Here, we present a protocol for immunofluorescent staining of blood and lymphatic endothelial markers in mouse whole-mount anterior eye cup at different postnatal ages. We describe steps for tissue harvesting, fixation, antibody incubation, mounting, and high-resolution confocal microscopy. This protocol facilitates the understanding of SC formation and functionality at the cellular level.For complete details on the use and execution of this protocol, please refer to Gu et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725003120metabolismmicroscopymolecular biologymolecular/chemical probes |
| spellingShingle | Weifa Wang Peiyao Peng Wenfo Hu Xuyang Liu Liming Yang Rong Ju Feng Zhang Protocol for immunofluorescent staining, high-resolution imaging, and spatial projection of Schlemm’s canal in mouse whole-mount corneal limbus tissue STAR Protocols metabolism microscopy molecular biology molecular/chemical probes |
| title | Protocol for immunofluorescent staining, high-resolution imaging, and spatial projection of Schlemm’s canal in mouse whole-mount corneal limbus tissue |
| title_full | Protocol for immunofluorescent staining, high-resolution imaging, and spatial projection of Schlemm’s canal in mouse whole-mount corneal limbus tissue |
| title_fullStr | Protocol for immunofluorescent staining, high-resolution imaging, and spatial projection of Schlemm’s canal in mouse whole-mount corneal limbus tissue |
| title_full_unstemmed | Protocol for immunofluorescent staining, high-resolution imaging, and spatial projection of Schlemm’s canal in mouse whole-mount corneal limbus tissue |
| title_short | Protocol for immunofluorescent staining, high-resolution imaging, and spatial projection of Schlemm’s canal in mouse whole-mount corneal limbus tissue |
| title_sort | protocol for immunofluorescent staining high resolution imaging and spatial projection of schlemm s canal in mouse whole mount corneal limbus tissue |
| topic | metabolism microscopy molecular biology molecular/chemical probes |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725003120 |
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