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.