A Procedure for Staining Cartilage and Bone of Whole Vertebrate Larvae While Rendering All Other Tissues Transparent

A new procedure for clearing and differentially staining skeletons of vertebrate larvae is described. Body pigments of fixed specimens are bleached by treatment with alkaline H₂O₂. This process markedly enhances the transparency of alcian blue-alizarine red S stained whole animals without damaging the specimens. This procedure should be useful for analyzing skeletal structures in studies of embryology, comparative anatomy and teratology of small vertebrates.