Expansion microscopy of the chick embryo neural tube to overcome molecular crowding at the centrosomes-cilia

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, a...

Descripción completa

Detalles Bibliográficos
Autores principales: Wilmerding, Axelle, Espana-Bonilla, Paula, Giakoumakis, Nikolaos-Nikiforos, Saade, Murielle
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9850183/
https://www.ncbi.nlm.nih.gov/pubmed/36609151
http://dx.doi.org/10.1016/j.xpro.2022.101997
_version_ 1784872126340136960
author Wilmerding, Axelle
Espana-Bonilla, Paula
Giakoumakis, Nikolaos-Nikiforos
Saade, Murielle
author_facet Wilmerding, Axelle
Espana-Bonilla, Paula
Giakoumakis, Nikolaos-Nikiforos
Saade, Murielle
author_sort Wilmerding, Axelle
collection PubMed
description 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.
format Online
Article
Text
id pubmed-9850183
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-98501832023-01-20 Expansion microscopy of the chick embryo neural tube to overcome molecular crowding at the centrosomes-cilia Wilmerding, Axelle Espana-Bonilla, Paula Giakoumakis, Nikolaos-Nikiforos Saade, Murielle STAR Protoc Protocol 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. Elsevier 2023-01-05 /pmc/articles/PMC9850183/ /pubmed/36609151 http://dx.doi.org/10.1016/j.xpro.2022.101997 Text en © 2023 The Authors. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Wilmerding, Axelle
Espana-Bonilla, Paula
Giakoumakis, Nikolaos-Nikiforos
Saade, Murielle
Expansion microscopy of the chick embryo neural tube to overcome molecular crowding at the centrosomes-cilia
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 Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9850183/
https://www.ncbi.nlm.nih.gov/pubmed/36609151
http://dx.doi.org/10.1016/j.xpro.2022.101997
work_keys_str_mv AT wilmerdingaxelle expansionmicroscopyofthechickembryoneuraltubetoovercomemolecularcrowdingatthecentrosomescilia
AT espanabonillapaula expansionmicroscopyofthechickembryoneuraltubetoovercomemolecularcrowdingatthecentrosomescilia
AT giakoumakisnikolaosnikiforos expansionmicroscopyofthechickembryoneuraltubetoovercomemolecularcrowdingatthecentrosomescilia
AT saademurielle expansionmicroscopyofthechickembryoneuraltubetoovercomemolecularcrowdingatthecentrosomescilia

Ejemplares similares