2021 article

Phenotyping Intact Mouse Bones Using Bone CLARITY

SKELETAL DEVELOPMENT AND REPAIR, 2 EDITION, Vol. 2230, pp. 217–230.

By: J. Treweek*, A. Beres n, N. Johnson  n & A. Greenbaum n

co-author countries: United States of America πŸ‡ΊπŸ‡Έ
author keywords: Tissue Clearing; Bone CLARITY; Three-dimensional imaging; Light-sheet microscopy; Fluorescence microscopy
MeSH headings : Animals; Bone and Bones / diagnostic imaging; Bone and Bones / physiology; Heme / chemistry; Humans; Hydrogels / pharmacology; Imaging, Three-Dimensional / methods; Mice; Optical Imaging / methods; Phenotype; Skull / diagnostic imaging; Skull / physiology; Staining and Labeling / methods
Source: Web Of Science
Added: August 9, 2021

The bone is typically studied using traditional histology techniques, that is, serial sectioning and staining. While effective, these techniques are laborious and destructive, as the native 3D environment of the bone is not maintained. Presented here is a bone-clearing methodology, termed Bone CLARITY, which combines published techniques for clearing soft tissues, including delipidation for the removal of light-scattering membranes, hydrogel-embedding for the stabilization of fragile epitopes, heme elution for the reduction of blood-based autofluorescence; as well as specialized steps, including decalcification and progressive refractive index matching, for addressing the unique challenges posed by osseous tissue. This method renders the bone transparent and enables the detailed visualization of an intact tissue specimen at multiple spatial scales.