The chemistry of fish ear bones (otoliths) is used to address fundamental questions in fish ecology and fisheries science. It is assumed that strontium (Sr), the most important element used in otolith chemistry research, is bound within the aragonitic calcium carbonate lattice of otoliths via random chemical replacement of calcium; however, this has never been tested and three other alternatives exist with regard to how Sr may be incorporated. If any variation in the mode of incorporation occurs, otolith chemistry data may be misinterpreted, impacting how fish and fisheries are understood and managed. Using X-ray absorption spectroscopy (specifically, analysis of extended X-ray absorption fine structure or EXAFS), we investigated how Sr is incorporated within fish otoliths from seven species collected from a range of aquatic environments. For comparison, aragonitic structures from other aquatic taxa (cephalopods and coral) were also analyzed. The results consistently indicated for all samples that Sr randomly replaces Ca within the aragonite lattice. This research explicitly shows how Sr is bound within otoliths and validates a fundamental and long-held assumption in aquatic research. ; Zoë A. Doubleday, Hugh H. Harris, Christopher Izzo, and Bronwyn M. Gillanders