A Protocol for Aging Anurans Using Skeletochronology

Age distribution information can be an important part of understanding the biology of any population. Age estimates collected from the annual growth rings found in tooth and bone cross sections, often referred to as Lines of Arrested Growth (LAGs), have been used in the study of various animals. In this manual, we describe in detail all necessary steps required to obtain estimates of age from anuran bone cross sections via skeletochronological assessment. We include comprehensive descriptions of how to fix and decalcify toe specimens (phalanges), process a phalange prior to embedding, embed the phalange in paraffin, section the phalange using a microtome, stain and mount the cross sections of the phalange and read the LAGs to obtain age estimates.