About the Applicability of Bone and Cartilage Acid-free Staining for Anura
The article describes the first use of acid-free alcian-alizarin staining for tailless amphibians in comparison with the standard method of acid staining on the example of the moor frog. The acid-free protocol is applicable for Anura and it has a number of advantages, such as the absence of decalcification, safety for the researcher and others. However, this method also has limitations, such as the binding of alcian blue with soft tissues. ; The authors thank Dmitriy Berzin and Natalia Ivanova for their help with growing tadpoles, Maxim Kovrizhin and Julia Sanaeva for their help with the chemical calculations, Yulia Kruzhkova for advice on the acid method of staining, Artem A. Minin, Artem S. Minin, Victor Sapronov, Ivan Sitnikov and Olga Zagainova for their help with the equipment and reagents and Vladimir Vershinin for his helpful comments and moral support. ; The work was performed within the frameworks of state contract with the Institute of Plant and Animal Ecology, Ural Branch, Russian Academy of Sciences, and partly supported by complex program of UBRAS (project № 18-4-4-28) and by Act 211, Government of the Russian Federation, contract № 02.A03.21.0006.