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...

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Detalles Bibliográficos
Autores: Wilmerding, Axelle, España-Bonilla, Paula, Giakoumakis, Nikolaos-Nikiforos, Saade, Murielle
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2023
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/335570
Acceso en línea:http://hdl.handle.net/10261/335570
Access Level:acceso abierto
Palabra clave:Cell biology
Developmental Biology
Microscopy
Descripción
Sumario: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.