Objetive. Human teeth have been commonly used for in vitro and in situ studies. Cu­rrently, other animals’ teeth have been purposed for dental research to overcome human teeth’ problematic availability. This study aimed to investigate the enamel and dentin from human, bovine, and ovine teeth concerning the microhardness, organic, and inorganic contents via micro-Raman spectroscopy. Methods. Human, bovine, and ovine teeth were divided according to their type and age into seven groups: Ovine; Bovine-12 months; Bovine-24 months; Bovine-36 months; Bovine-48 months; Bovine-+60 months; Human (control). The enamel’s microhardness (superficial and deep) and dentin (superficial, middle, and deep) were analyzed. The calcium/phosphate ratio and amide contents were determined by micro-Raman spectroscopy. Results. Overall, the microhardness of human enamel was superior to the other species. De...