Decalcifying odontocete ears following a routine protocol with RDO®

Abstract

The study of the organ of Corti is essential to assess the impact of underwater noise on cetaceans. While classical histology techniques (including EDTA decalcification) have been previously considered, the process is time consuming. Independently from the histological technique, one of the challenging steps after extraction and fixation of the samples is to decalcify the bone envelope to access the cochlea without damaging the soft tissues. Here, we propose to use a fast commercial decalcifier (RDO®). 93 ears from 11 different odontocetes species stranded in the Mediterranean, Spanish North Atlantic and North Sea were used to precisely determine the decalcification time. Depending on the tympanic–periotic volume of the species, the decalcification time ranged from several hours to a few days, allowing a subsequently faster observation of the cochlear structures through routine microscope techniques.