Publication Date

6-21-2024

Journal

STAR Protocols

DOI

10.1016/j.xpro.2024.103053

PMID

38704833

PMCID

PMC11089399

PubMedCentral® Posted Date

5-4-2024

PubMedCentral® Full Text Version

Post-print

Published Open-Access

yes

Keywords

Animals, Mice, Imaging, Three-Dimensional, Fluorescent Dyes, Optical Imaging, Developmental biology, microscopy, Model Organisms, Neuroscience

Abstract

Tissue clearing is an essential prerequisite for 3D volumetric imaging of larger tissues or organs. Here, we present a detailed protocol for optical, aqueous-based clearing of adult murine tissues using EZ Clear. We describe steps to ensure successful perfusion and fixation of organs from the adult mouse and supply guidelines for optimal lipid removal, refractive index matching, and tissue clearing. Finally, we provide imaging parameters for visualizing both exogenous perfused fluorescent dyes and endogenous fluorescence reporters in the adult mouse. For complete details on the use and execution of this protocol, please refer to Hsu et al.

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