Light and focused ion beam microscopy workflow for resin-embedded tissues
Although the automated image acquisition with the focused ion beam scanning electron microscope (FIB-SEM) provides volume reconstructions, volume analysis of large samples remains challenging. Here, we present a workflow that combines a modified sample protocol of the classical transmission electron...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-01-01
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| Series: | Frontiers in Cell and Developmental Biology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2023.1076736/full |
| _version_ | 1797943607627350016 |
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| author | Horacio Merchant-Larios David M. Giraldo-Gomez Adriana Castro-Dominguez Alejandro Marmolejo-Valencia |
| author_facet | Horacio Merchant-Larios David M. Giraldo-Gomez Adriana Castro-Dominguez Alejandro Marmolejo-Valencia |
| author_sort | Horacio Merchant-Larios |
| collection | DOAJ |
| description | Although the automated image acquisition with the focused ion beam scanning electron microscope (FIB-SEM) provides volume reconstructions, volume analysis of large samples remains challenging. Here, we present a workflow that combines a modified sample protocol of the classical transmission electron microscope with FIB-SEM volume imaging. The proposed workflow enables efficient 3D structural surveys of rabbit ovaries collected at consecutive developmental stages. The precise trimming of the region of interest adds the time dimension to the volume, constructing a virtual 4D electron microscopy. We found filopodia-like processes emitted by oocyte cysts allowing contact between oocytes not previously observed. |
| first_indexed | 2024-04-10T20:26:51Z |
| format | Article |
| id | doaj.art-d517ab34860d4f588ff7f966083a253b |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-04-10T20:26:51Z |
| publishDate | 2023-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-d517ab34860d4f588ff7f966083a253b2023-01-25T10:49:02ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2023-01-011110.3389/fcell.2023.10767361076736Light and focused ion beam microscopy workflow for resin-embedded tissuesHoracio Merchant-Larios0David M. Giraldo-Gomez1Adriana Castro-Dominguez2Alejandro Marmolejo-Valencia3Instituto de Investigaciones Biomédicas Departamento de Biología Celular y Fisiología, Universidad Nacional Autónoma de México (UNAM), México City, MéxicoCarl Zeiss de México S. A. de C. V., Research Microscopy Solutions, , México City, MexicoInstituto de Investigaciones Biomédicas Departamento de Biología Celular y Fisiología, Universidad Nacional Autónoma de México (UNAM), México City, MéxicoInstituto de Investigaciones Biomédicas Departamento de Biología Celular y Fisiología, Universidad Nacional Autónoma de México (UNAM), México City, MéxicoAlthough the automated image acquisition with the focused ion beam scanning electron microscope (FIB-SEM) provides volume reconstructions, volume analysis of large samples remains challenging. Here, we present a workflow that combines a modified sample protocol of the classical transmission electron microscope with FIB-SEM volume imaging. The proposed workflow enables efficient 3D structural surveys of rabbit ovaries collected at consecutive developmental stages. The precise trimming of the region of interest adds the time dimension to the volume, constructing a virtual 4D electron microscopy. We found filopodia-like processes emitted by oocyte cysts allowing contact between oocytes not previously observed.https://www.frontiersin.org/articles/10.3389/fcell.2023.1076736/fullfocused ion beam3D tissue ultrastructureresin embeddedlarge samplesdeveloping mammalian ovaries |
| spellingShingle | Horacio Merchant-Larios David M. Giraldo-Gomez Adriana Castro-Dominguez Alejandro Marmolejo-Valencia Light and focused ion beam microscopy workflow for resin-embedded tissues Frontiers in Cell and Developmental Biology focused ion beam 3D tissue ultrastructure resin embedded large samples developing mammalian ovaries |
| title | Light and focused ion beam microscopy workflow for resin-embedded tissues |
| title_full | Light and focused ion beam microscopy workflow for resin-embedded tissues |
| title_fullStr | Light and focused ion beam microscopy workflow for resin-embedded tissues |
| title_full_unstemmed | Light and focused ion beam microscopy workflow for resin-embedded tissues |
| title_short | Light and focused ion beam microscopy workflow for resin-embedded tissues |
| title_sort | light and focused ion beam microscopy workflow for resin embedded tissues |
| topic | focused ion beam 3D tissue ultrastructure resin embedded large samples developing mammalian ovaries |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2023.1076736/full |
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