Open AccessUrea-based amino sugar agent clears murine liver and preserves protein fluorescence and lipophilic dyes
Author(s) -
Michelle Hough,
Michael Fenlon,
Alison Glazier,
Celia Short,
G. Esteban Fernández,
Jiabo Xu,
Elaa Mahdi,
Kinji Asahina,
Kasper S. Wang
Publication year - 2021
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2020-0063
Subject(s) - reagent , fluorophore , urea , fluorescence , fluorescent protein , biochemistry , sugar , chemistry , endogeny , labelling , amino acid , reporter gene , biology , chromatography , green fluorescent protein , gene , gene expression , organic chemistry , physics , quantum mechanics
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.