Protocol for optical, aqueous-based clearing of murine tissues using EZ Clear
Taeyong Ahn, Gabrielle Largoza, Julia Younis, Mary E. Dickinson, Chih‐Wei Hsu, Joshua D. Wythe
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.1
Topics & Concepts
FluorescenceClearingFixation (population genetics)Protocol (science)Biomedical engineeringEndogenyBiophysicsChemistryComputer scienceCell biologyBiologyPathologyBiochemistryMedicineOpticsAlternative medicinePhysicsGeneFinanceEconomicsAdvanced Fluorescence Microscopy TechniquesCell Image Analysis TechniquesSingle-cell and spatial transcriptomics