EZ Clear for Simple, Rapid, and Robust Mouse Whole Organ Clearing

Authors

Chih-Wei Hsu
Juan Cerda
Jason M Kirk
Williamson D Turner
Tara L Rasmussen
Carlos P Flores Suarez
Mary E Dickinson
Joshua D Wythe

Publication Date

10-11-2022

Journal

ELife

DOI

10.7554/eLife.77419

PMID

36218247

PMCID

PMC9555867

PubMedCentral® Posted Date

10-11-2022

PubMedCentral® Full Text Version

Post-print

Published Open-Access

yes

Keywords

Animals, Brain, Coloring Agents, Imaging, Three-Dimensional, Mice, Solvents, Staining and Labeling

Abstract

Tissue clearing for whole organ cell profiling has revolutionized biology and imaging for exploration of organs in three-dimensional space without compromising tissue architecture. But complicated, laborious procedures, or expensive equipment, as well as the use of hazardous, organic solvents prevent the widespread adoption of these methods. Here, we report a simple and rapid tissue clearing method, EZ Clear, that can clear whole adult mouse organs in 48 hr in just three simple steps. Samples stay at room temperature and remain hydrated throughout the clearing process, preserving endogenous and synthetic fluorescence, without altering sample size. After wholemount clearing and imaging, samples processed with EZ Clear can be subjected to downstream applications, such as tissue embedding and cryosectioning followed by standard histology or immunofluorescent staining without loss of fluorescence signal from endogenous or synthetic reporters. Furthermore, we demonstrate that wholemount adult mouse brains processed with EZ Clear can be successfully immunolabeled for fluorescent imaging while still retaining signal from endogenous fluorescent reporters. Overall, the simplicity, speed, and flexibility of EZ Clear make it easy to adapt and implement in diverse imaging modalities in biomedical research.

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