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Updated: May 17, 2026

Undecalcified Bone Preparation for Histology, Histomorphometry and Fluorochrome Analysis
Published on: January 8, 2010
Skeletal examination by double staining for ossified bone and cartilaginous tissue
1BASF SE, Ludwigshafen, Germany. steffen.schneider@basf.com
Abstract:
The assessment of developmental toxicology data is a critical aspect of hazard evaluation for pharmaceuticals and environmental chemicals. Skeletal examination is an essential part of prenatal developmental toxicity studies of chemicals as well as pesticides and comprises evaluation of both cartilaginous and ossified skeletal components. Various techniques are published in the literature to process and double-stain skeletons of common laboratory animals which are all based on staining of the cartilage with Alcian Blue and staining of ossified bones with Alizarin Red S along with maceration (clearing) of the surrounding soft tissue. The staining of the cartilage allows the examination to assess ossified structures and their underlying cartilage in a single step.
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