Expansion microscopy of the chick embryo neural tube to overcome molecular crowding at the centrosomes-cilia
Summary: We describe an optimized protocol for application of expansion microscopy (ExM) on chick neural tube (NT) which enables different oriented nanoscale resolution imaging of the centrosomes/cilia. We explain embryo NT transversal sections and open-book preparations, immunohistochemistry for la...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722008772 |
| _version_ | 1828070228036157440 |
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| author | Axelle Wilmerding Paula Espana-Bonilla Nikolaos-Nikiforos Giakoumakis Murielle Saade |
| author_facet | Axelle Wilmerding Paula Espana-Bonilla Nikolaos-Nikiforos Giakoumakis Murielle Saade |
| author_sort | Axelle Wilmerding |
| collection | DOAJ |
| description | Summary: We describe an optimized protocol for application of expansion microscopy (ExM) on chick neural tube (NT) which enables different oriented nanoscale resolution imaging of the centrosomes/cilia. We explain embryo NT transversal sections and open-book preparations, immunohistochemistry for labeling, and sample preparation for 5-fold tissue expansion. Further, we detail sample orientation and Fast Airyscan confocal acquisition and show that NT-ExM retains fluorescence signals and overcomes biomolecules crowding in structural features that to date were only imaged with electron microscopy on tissues. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-11T00:33:33Z |
| format | Article |
| id | doaj.art-285db0b8b893400ca980f6f37e3237c4 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-11T00:33:33Z |
| publishDate | 2023-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-285db0b8b893400ca980f6f37e3237c42023-01-07T04:17:24ZengElsevierSTAR Protocols2666-16672023-03-0141101997Expansion microscopy of the chick embryo neural tube to overcome molecular crowding at the centrosomes-ciliaAxelle Wilmerding0Paula Espana-Bonilla1Nikolaos-Nikiforos Giakoumakis2Murielle Saade3Department of Cells and Tissues, Instituto de Biología Molecular de Barcelona, Consejo Superior de Investigaciones Científicas (IBMB-CSIC), Parc Científic de Barcelona, C/Baldiri i Reixac 20, 08028 Barcelona, SpainDepartment of Cells and Tissues, Instituto de Biología Molecular de Barcelona, Consejo Superior de Investigaciones Científicas (IBMB-CSIC), Parc Científic de Barcelona, C/Baldiri i Reixac 20, 08028 Barcelona, SpainAdvanced Digital Microscopy Facility Institute for Research in Biomedicine – IRB, Parc Científic de Barcelona, C/Baldiri Reixac 10, 08028 Barcelona, Spain; Corresponding authorDepartment of Cells and Tissues, Instituto de Biología Molecular de Barcelona, Consejo Superior de Investigaciones Científicas (IBMB-CSIC), Parc Científic de Barcelona, C/Baldiri i Reixac 20, 08028 Barcelona, Spain; Corresponding authorSummary: We describe an optimized protocol for application of expansion microscopy (ExM) on chick neural tube (NT) which enables different oriented nanoscale resolution imaging of the centrosomes/cilia. We explain embryo NT transversal sections and open-book preparations, immunohistochemistry for labeling, and sample preparation for 5-fold tissue expansion. Further, we detail sample orientation and Fast Airyscan confocal acquisition and show that NT-ExM retains fluorescence signals and overcomes biomolecules crowding in structural features that to date were only imaged with electron microscopy on tissues. : 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/S2666166722008772Cell BiologyDevelopmental BiologyMicroscopy |
| spellingShingle | Axelle Wilmerding Paula Espana-Bonilla Nikolaos-Nikiforos Giakoumakis Murielle Saade Expansion microscopy of the chick embryo neural tube to overcome molecular crowding at the centrosomes-cilia STAR Protocols Cell Biology Developmental Biology Microscopy |
| title | Expansion microscopy of the chick embryo neural tube to overcome molecular crowding at the centrosomes-cilia |
| title_full | Expansion microscopy of the chick embryo neural tube to overcome molecular crowding at the centrosomes-cilia |
| title_fullStr | Expansion microscopy of the chick embryo neural tube to overcome molecular crowding at the centrosomes-cilia |
| title_full_unstemmed | Expansion microscopy of the chick embryo neural tube to overcome molecular crowding at the centrosomes-cilia |
| title_short | Expansion microscopy of the chick embryo neural tube to overcome molecular crowding at the centrosomes-cilia |
| title_sort | expansion microscopy of the chick embryo neural tube to overcome molecular crowding at the centrosomes cilia |
| topic | Cell Biology Developmental Biology Microscopy |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722008772 |
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