Urea-based amino sugar agent clears murine liver and preserves protein fluorescence and lipophilic dyes

Five established clearing protocols were compared with a modified and simplified method to determine an optimal clearing reagent for three-dimensionally visualizing fluorophores in the murine liver, a challenging organ to clear. We report successful clearing of whole liver lobes by modification of a...

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Veröffentlicht in:BioTechniques. - 1991. - 70(2021), 2 vom: 25. Feb., Seite 72-80
1. Verfasser: Hough, Michelle (VerfasserIn)
Weitere Verfasser: Fenlon, Michael, Glazier, Alison, Short, Celia, Fernandez, Gerardo Esteban, Xu, Jiabo, Mahdi, Elaa, Asahina, Kinji, Wang, Kasper S
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2021
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte:Journal Article Research Support, N.I.H., Extramural 3D imaging bile duct ligation computational imaging fluorescence microscopy lipophilic tracers liver clearing optical clearing agents prominin-1 mehr... Amino Sugars Fluorescent Dyes Urea 8W8T17847W
Beschreibung
Zusammenfassung:Five established clearing protocols were compared with a modified and simplified method to determine an optimal clearing reagent for three-dimensionally visualizing fluorophores in the murine liver, a challenging organ to clear. We report successful clearing of whole liver lobes by modification of an established protocol (UbasM) using only Ub-1, a urea-based amino sugar reagent, in a simpler protocol that requires only a 24-h processing time. With Ub-1 alone, we observed sufficiently preserved liver tissue structure in three dimensions along with excellent preservation of fluorophore emissions from endogenous protein reporters and lipophilic tracer dyes. This streamlined technique can be used for 3D cell lineage tracing and fluoroprobe-based reporter gene expression to compare various experimental conditions
Beschreibung:Date Completed 06.12.2021
Date Revised 14.12.2021
published: Print-Electronic
Citation Status MEDLINE
ISSN:1940-9818
DOI:10.2144/btn-2020-0063