Urea-Based Amino Sugar Agent Clears Murine Liver And Preserves Protein Fluorescence And Lipophilic Dyes

METHOD SUMMARYThis study presents a simplified protocol for optically clearing murine liver tissue in only 24 h using one simple urea-based amino sugar solution and a single incubation. This method preserves fluorescence of transgenically expressed proteins and lipophilic tracer dyes within the context of native spatial morphology.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.