Protocol for assessing regional pathology in the rodent optic nerve using longitudinal cryosections and cross-sectional electron microscopy
Summary: Visual system pathology has been established as a prominent feature in demyelinating diseases such as multiple sclerosis (MS). Here, we present a protocol for assessing regional pathology in the rodent optic nerve using longitudinal cryosections and cross-sectional electron microscopy. We d...
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| Language: | English |
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Elsevier
2025-06-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725002667 |
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| author | Gabrielle M. Mey John Shelestak Muhammad Irfan Tara M. DeSilva |
| author_facet | Gabrielle M. Mey John Shelestak Muhammad Irfan Tara M. DeSilva |
| author_sort | Gabrielle M. Mey |
| collection | DOAJ |
| description | Summary: Visual system pathology has been established as a prominent feature in demyelinating diseases such as multiple sclerosis (MS). Here, we present a protocol for assessing regional pathology in the rodent optic nerve using longitudinal cryosections and cross-sectional electron microscopy. We describe steps for optic nerve dissection, post-fixation, myelin detection, slide scanning of myelin basic protein, and the analysis of demyelination. We then detail procedures for fluorescent antibody staining, analysis for transmission electron microscopy (TEM), and the analysis of myelinated and unmyelinated axons.For complete details on the use and execution of this protocol, please refer to Mey et al.1,2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-33fbe366dbc74133aa93a7c0863c1df4 |
| institution | DOAJ |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-33fbe366dbc74133aa93a7c0863c1df42025-08-20T03:07:51ZengElsevierSTAR Protocols2666-16672025-06-016210386010.1016/j.xpro.2025.103860Protocol for assessing regional pathology in the rodent optic nerve using longitudinal cryosections and cross-sectional electron microscopyGabrielle M. Mey0John Shelestak1Muhammad Irfan2Tara M. DeSilva3Department of Ophthalmology and Visual Sciences, University of Michigan, Michigan Medicine, Ann Arbor, MI 48105, USA; Michigan Neuroscience Institute, University of Michigan, Michigan Medicine, Ann Arbor, MI 48105, USADepartment of Neurosciences, Cleveland Clinic Lerner College of Medicine and Case Western Reserve University, Cleveland, OH 44195, USADepartment of Neurosciences, Cleveland Clinic Lerner College of Medicine and Case Western Reserve University, Cleveland, OH 44195, USADepartment of Neurosciences, Cleveland Clinic Lerner College of Medicine and Case Western Reserve University, Cleveland, OH 44195, USA; Corresponding authorSummary: Visual system pathology has been established as a prominent feature in demyelinating diseases such as multiple sclerosis (MS). Here, we present a protocol for assessing regional pathology in the rodent optic nerve using longitudinal cryosections and cross-sectional electron microscopy. We describe steps for optic nerve dissection, post-fixation, myelin detection, slide scanning of myelin basic protein, and the analysis of demyelination. We then detail procedures for fluorescent antibody staining, analysis for transmission electron microscopy (TEM), and the analysis of myelinated and unmyelinated axons.For complete details on the use and execution of this protocol, please refer to Mey et al.1,2 : 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/S2666166725002667Cell biologyMicroscopyNeuroscience |
| spellingShingle | Gabrielle M. Mey John Shelestak Muhammad Irfan Tara M. DeSilva Protocol for assessing regional pathology in the rodent optic nerve using longitudinal cryosections and cross-sectional electron microscopy STAR Protocols Cell biology Microscopy Neuroscience |
| title | Protocol for assessing regional pathology in the rodent optic nerve using longitudinal cryosections and cross-sectional electron microscopy |
| title_full | Protocol for assessing regional pathology in the rodent optic nerve using longitudinal cryosections and cross-sectional electron microscopy |
| title_fullStr | Protocol for assessing regional pathology in the rodent optic nerve using longitudinal cryosections and cross-sectional electron microscopy |
| title_full_unstemmed | Protocol for assessing regional pathology in the rodent optic nerve using longitudinal cryosections and cross-sectional electron microscopy |
| title_short | Protocol for assessing regional pathology in the rodent optic nerve using longitudinal cryosections and cross-sectional electron microscopy |
| title_sort | protocol for assessing regional pathology in the rodent optic nerve using longitudinal cryosections and cross sectional electron microscopy |
| topic | Cell biology Microscopy Neuroscience |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725002667 |
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