The elemental composition of fish otoliths is of considerable interest to those who wish to reconstruct temperature, migration, or environmental histories of individual fish based on assays of the otolith growth sequence. However, reported differences in otolith elemental composition among studies may be due in part to performance differences among four of the most popular instruments for targeted elemental analysis: wavelength-dispersive electron microprobe (WD-EM), energy-dispersive electron microprobe (ED-EM), proton-induced X-ray emission (PIXE), and laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS). To rigorously compare the sensitivity, accuracy, and precision of these four analytical tools, the International Otolith Composition Experiment distributed blind-labelled real and artificial otoliths of known but varied elemental composition to eight laboratories for assay of 10 selected elements. No one instrument type was sensitive to each element, nor was any one instrument preferred for use in all assays. In general however, abundant elements such as Na and K could only be measured accurately with an electron microprobe, while the trace elements required PIXE or LA-ICPMS. Strontium could be measured with considerable accuracy and precision by WD-EM, PIXE, and LA-ICPMS. The presence of significant, and occasionally large, differences among laboratories suggests that comparisons among published studies should be made cautiously and only after appropriate calibration.