The aim of this study was to assess the suitability of different vertebrae staining techniques for the visualization and counting of growth bands in tropical species of batoids (Narcine leoparda, Urotrygon aspidura, Hypanus longus, Potamotrygon magdalenae) and sharks (Alopias pelagicus, Carcharhinus falciformis, Sphyrna lewini, Sphyrna corona and Mustelus lunulatus). Different cutting thicknesses and staining protocols were tested, analysing the precision and bias of each combination to identify the most accurate technique for estimating age. Vertebral sections of 0.4 mm were more suitable for batoids, except for Narcine leoparda; for this species and for all the shark species assessed, sections of 0.5 mm are recommended. Different combinations of stain and exposure time were required to achieve the best visualizations of vertebral growth band pair for the shark and ray species. Intraspecific variation occurred among vertebrae size of batoids. Our results confirm the importance of defining a suitable species-specific protocol for sectioning and staining hard structures before carrying out an age and growth study to improve the reliability of the age estimates.