Urea-based amino sugar agent clears murine liver and preserves protein fluorescence and lipophilic dyes
Michelle Hough,
Michael Fenlon,
Alison Glazier,
Celia Short,
Gerardo Esteban Fernandez,
Jiabo Xu,
Elaa Mahdi,
Kinji Asahina,
Kasper S Wang
Affiliations
Michelle Hough
1Developmental Biology, Regenerative Medicine & Stem Cell Program, The Saban Research Institute, Children’s Hospital of Los Angeles, Los Angeles, CA 90027, USA
Michael Fenlon
1Developmental Biology, Regenerative Medicine & Stem Cell Program, The Saban Research Institute, Children’s Hospital of Los Angeles, Los Angeles, CA 90027, USA
Alison Glazier
1Developmental Biology, Regenerative Medicine & Stem Cell Program, The Saban Research Institute, Children’s Hospital of Los Angeles, Los Angeles, CA 90027, USA
Celia Short
1Developmental Biology, Regenerative Medicine & Stem Cell Program, The Saban Research Institute, Children’s Hospital of Los Angeles, Los Angeles, CA 90027, USA
Gerardo Esteban Fernandez
1Developmental Biology, Regenerative Medicine & Stem Cell Program, The Saban Research Institute, Children’s Hospital of Los Angeles, Los Angeles, CA 90027, USA
Jiabo Xu
1Developmental Biology, Regenerative Medicine & Stem Cell Program, The Saban Research Institute, Children’s Hospital of Los Angeles, Los Angeles, CA 90027, USA
Elaa Mahdi
1Developmental Biology, Regenerative Medicine & Stem Cell Program, The Saban Research Institute, Children’s Hospital of Los Angeles, Los Angeles, CA 90027, USA
Kinji Asahina
2Southern California Research Center for ALPD & Cirrhosis, Department of Pathology, Keck School of Medicine, University of Southern California, Los Angeles, CA 90089, USA
Kasper S Wang
1Developmental Biology, Regenerative Medicine & Stem Cell Program, The Saban Research Institute, Children’s Hospital of Los Angeles, Los Angeles, CA 90027, USA
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.
