High-resolution imaging of fluorescent whole mouse brains using stabilised organic media (sDISCO).
EGFP
fluorescence stabilisation
light-sheet microscopy
mouse brain
optical tissue clearing
ultramicroscopy
whole-tissue imaging
Journal
Journal of biophotonics
ISSN: 1864-0648
Titre abrégé: J Biophotonics
Pays: Germany
ID NLM: 101318567
Informations de publication
Date de publication:
08 2019
08 2019
Historique:
received:
24
09
2018
revised:
26
03
2019
accepted:
28
03
2019
pubmed:
2
4
2019
medline:
15
9
2020
entrez:
2
4
2019
Statut:
ppublish
Résumé
Optical tissue clearing using dibenzyl ether (DBE) or BABB (1 part benzyl alcohol and 2 parts benzyl benzoate) is easy in application and allows deep-tissue imaging of a wide range of specimens. However, in both substances, optical clearing and storage times of enhanced green fluorescent protein (EGFP)-expressing specimens are limited due to the continuous formation of peroxides and aldehydes, which severely quench fluorescence. Stabilisation of purified DBE or BABB by addition of the antioxidant propyl gallate efficiently preserves fluorescence signals in EGFP-expressing samples for more than a year. This enables longer clearing times and improved tissue transparency with higher fluorescence signal intensity. The here introduced clearing protocol termed stabilised DISCO allows to image spines in a whole mouse brain and to detect faint changes in the activity-dependent expression pattern of tdTomato.
Identifiants
pubmed: 30932329
doi: 10.1002/jbio.201800368
doi:
Substances chimiques
enhanced green fluorescent protein
0
Green Fluorescent Proteins
147336-22-9
Types de publication
Editorial
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e201800368Subventions
Organisme : Austrian Science Fund FWF
ID : P 28338
Pays : Austria
Organisme : FWF
ID : P23102-N22
Pays : International
Informations de copyright
© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.