Multiplexed volumetric CLEM enabled by scFvs provides insights into the cytology of cerebellar cortex.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
05 Aug 2024
05 Aug 2024
Historique:
received:
28
06
2023
accepted:
01
07
2024
medline:
6
8
2024
pubmed:
6
8
2024
entrez:
5
8
2024
Statut:
epublish
Résumé
Mapping neuronal networks is a central focus in neuroscience. While volume electron microscopy (vEM) can reveal the fine structure of neuronal networks (connectomics), it does not provide molecular information to identify cell types or functions. We developed an approach that uses fluorescent single-chain variable fragments (scFvs) to perform multiplexed detergent-free immunolabeling and volumetric-correlated-light-and-electron-microscopy on the same sample. We generated eight fluorescent scFvs targeting brain markers. Six fluorescent probes were imaged in the cerebellum of a female mouse, using confocal microscopy with spectral unmixing, followed by vEM of the same sample. The results provide excellent ultrastructure superimposed with multiple fluorescence channels. Using this approach, we documented a poorly described cell type, two types of mossy fiber terminals, and the subcellular localization of one type of ion channel. Because scFvs can be derived from existing monoclonal antibodies, hundreds of such probes can be generated to enable molecular overlays for connectomic studies.
Identifiants
pubmed: 39103318
doi: 10.1038/s41467-024-50411-z
pii: 10.1038/s41467-024-50411-z
doi:
Substances chimiques
Fluorescent Dyes
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
6648Subventions
Organisme : NINDS NIH HHS
ID : U19 NS104653
Pays : United States
Organisme : NIMH NIH HHS
ID : UG3 MH123386
Pays : United States
Organisme : NIMH NIH HHS
ID : P50 MH094271
Pays : United States
Informations de copyright
© 2024. The Author(s).
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