Protocol for the three-dimensional analysis of rodent skeletal muscle
Summary: Confocal imaging is a powerful tool capable of analyzing cellular spatial data within a given tissue. Here, we present a protocol for preparing optically cleared extensor digitorum longus (EDL) skeletal muscle samples suitable for confocal imaging/computational analysis. We describe steps f...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724007147 |
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| author | Smrithi Karthikeyan Yoko Asakura Mayank Verma Atsushi Asakura |
| author_facet | Smrithi Karthikeyan Yoko Asakura Mayank Verma Atsushi Asakura |
| author_sort | Smrithi Karthikeyan |
| collection | DOAJ |
| description | Summary: Confocal imaging is a powerful tool capable of analyzing cellular spatial data within a given tissue. Here, we present a protocol for preparing optically cleared extensor digitorum longus (EDL) skeletal muscle samples suitable for confocal imaging/computational analysis. We describe steps for sample preparation (including perfusion fixation and tissue clearing of muscle samples), image acquisition, and computational analysis, with sample segmentation/3D rendering outlined. This protocol can be applied to characterize various cell types, including muscle satellite cells (muscle stem cells) and capillary endothelial cells within rodent skeletal muscle.For complete details on the use and execution of this protocol, please refer to Verma et al.,1 Verma et al.,2 Karthikeyan et al.,3 and Karthikeyan et al.4 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-c897e8b5d04c4d138c489c1918d8bbd4 |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-c897e8b5d04c4d138c489c1918d8bbd42025-01-11T06:41:59ZengElsevierSTAR Protocols2666-16672025-03-0161103549Protocol for the three-dimensional analysis of rodent skeletal muscleSmrithi Karthikeyan0Yoko Asakura1Mayank Verma2Atsushi Asakura3Stem Cell Institute, University of Minnesota Medical School, Minneapolis, MN, USA; Paul & Sheila Wellstone Muscular Dystrophy Center, University of Minnesota Medical School, Minneapolis, MN, USA; Department of Neurology, University of Minnesota Medical School, Minneapolis, MN, USAStem Cell Institute, University of Minnesota Medical School, Minneapolis, MN, USA; Paul & Sheila Wellstone Muscular Dystrophy Center, University of Minnesota Medical School, Minneapolis, MN, USA; Department of Neurology, University of Minnesota Medical School, Minneapolis, MN, USADepartment of Pediatrics & Neurology, Division of Pediatric Neurology, The University of Texas Southwestern Medical Center, Dallas, TX, USAStem Cell Institute, University of Minnesota Medical School, Minneapolis, MN, USA; Paul & Sheila Wellstone Muscular Dystrophy Center, University of Minnesota Medical School, Minneapolis, MN, USA; Department of Neurology, University of Minnesota Medical School, Minneapolis, MN, USA; Corresponding authorSummary: Confocal imaging is a powerful tool capable of analyzing cellular spatial data within a given tissue. Here, we present a protocol for preparing optically cleared extensor digitorum longus (EDL) skeletal muscle samples suitable for confocal imaging/computational analysis. We describe steps for sample preparation (including perfusion fixation and tissue clearing of muscle samples), image acquisition, and computational analysis, with sample segmentation/3D rendering outlined. This protocol can be applied to characterize various cell types, including muscle satellite cells (muscle stem cells) and capillary endothelial cells within rodent skeletal muscle.For complete details on the use and execution of this protocol, please refer to Verma et al.,1 Verma et al.,2 Karthikeyan et al.,3 and Karthikeyan et al.4 : 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/S2666166724007147classification Description: cell biologydevelopmental biologymodel organismsmolecular biologystem cellscell differentiation |
| spellingShingle | Smrithi Karthikeyan Yoko Asakura Mayank Verma Atsushi Asakura Protocol for the three-dimensional analysis of rodent skeletal muscle STAR Protocols classification Description: cell biology developmental biology model organisms molecular biology stem cells cell differentiation |
| title | Protocol for the three-dimensional analysis of rodent skeletal muscle |
| title_full | Protocol for the three-dimensional analysis of rodent skeletal muscle |
| title_fullStr | Protocol for the three-dimensional analysis of rodent skeletal muscle |
| title_full_unstemmed | Protocol for the three-dimensional analysis of rodent skeletal muscle |
| title_short | Protocol for the three-dimensional analysis of rodent skeletal muscle |
| title_sort | protocol for the three dimensional analysis of rodent skeletal muscle |
| topic | classification Description: cell biology developmental biology model organisms molecular biology stem cells cell differentiation |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724007147 |
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