The morphological and chemical changes in deciduous dentin produced by different conditioning protocols were evaluated in this in vitro study. Eighty primary dentin samples were divided into eight groups (n = 10): G1, acid etching; G2, self-etching adhesive; G3, G4, Er: YAG laser irradiation at 25.5 and 38.2 J cm , respectively; 10 Hz and spray irrigation. Groups 5, 6, 7, and 8 were irradiated at previous densities, and then phosphoric acid or self-etching adhesive conditioning was applied. Scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS) were used to evaluate chemical and morphological changes. Paired t-test and One-way ANOVA were used for statistical analysis (p ≤ 0.05). All samples showed different morphology with specific characteristics according to the conditioning protocol. Changing element concentration values are expressed in atomic percent (at %). After conditioning, there were statistically significant differences (p ≤ 0.05) for p at% and Ca/P in all groups; highlighting the following additional findings by group: G1, G7, and G8 showed changes in all elements studied, G2 presented a decrease in C at% and increased Ca at%, G3 and G4 exhibited at% changes in C, trace elements and Ca. Furthermore, G5 showed at% changes in O and trace elements; while G6 changes were observed on C at%, O at% and trace elements at%. Dentin morphology and chemical composition varied in accordance with the conditioning protocol, with characteristics specific for each one that could have clinical implications for the retention and bond strength performance of adhesive materials.