Mock double staining

Transparent skeletal specimens are generally prepared by staining ossified bones with alizarin red and cartilage with alcian blue, or using a double-staining technique, followed by the clarification of soft tissues. The vividly stained skeleton can be observed in its original three-dimensional arrangement within the transparent flesh, overcoming the aforementioned difficulties. Alizarin red is used in biological research to stain calcium salt deposits in living organisms due to its ability to bind with metal ions. Alcian blue is a basic dye belonging to the phthalocyanine group, which forms ionic bonds with the carboxyl and sulfate groups of acidic mucopolysaccharides.