Background. Enamel microabrasion can eliminate enamel irregularities and discoloration. This study evaluated the staining susceptibility of enamel after microabrasion using different clinical protocols. Methods. A total of 210 extracted bovine incisors were used in this study. The teeth were randomly divided into five groups of 42 teeth each (n=42), including group 1: control (no treatment), group 2: microabrasion, group 3: microabrasion + fluoride polishing, group 4: macroabrasion (fine-grit diamond bur) + microabrasion, and group 5: macroabrasion (finegrit diamond bur) + microabrasion + fluoride polishing. The groups were then randomly divided into two subgroups for discoloration procedures: coffee and distilled water (n=21). A spectrophotometric evaluation was carried out at baseline and on the 1st, 7th, 14th, and 28th days of the staining period. Statistical analyses were performed using repeated-measures ANOVA and the post hoc Bonferroni test at a significance level of 0.05. Results. The greatest color change was observed in group 2 specimens, which were immersed in coffee solutions. The color change values for groups 3, 4, and 5 specimens, which were immersed in distilled water, were higher than those in group 1 specimens (P<0.05). The fluoride polishing + enamel microabrasion treatment groups (groups 3 and 5) exhibited greater resistance to color changes than the matched non-fluoride-polished groups (groups 2 and 4) (P<0.05). Conclusions. The teeth that underwent enamel microabrasion treatment and were polished with fluoride gel became more resistant to color changes. Our findings confirm that enamel microabrasion treatment is a conservative method for localized discoloration.