Objective To observe the efficiency and range of collateral tissue damage in ablation of rat bladder epithelial tissues by femtosecond laser. Methods Bladder epithelial tissues of SD rats were ablated by femtosecond laser with different working powers (1, 2, 5, 10 and 20 times of threshold power and equivalent frequencies). Sections with HE staining were prepared for the ablated tissues. The depth and range of collateral tissue damage of bladder epithelial tissues ablated by femtosecond laser were observed by light microscopy, and the ablation efficiency was calculated. Results When the working powers of femtosecond laser were 5 to 20 times of threshold power and the equivalent frequencies were 62.5 Hz to 1 000 Hz, the ablation efficiency varied from 10 to 120 μm/s and the collateral tissue damage ranged from 10 to 100 μm. Conclusion Femtosecond laser can precisely ablate the rat bladder epithelial tissues with minimal range of collateral tissue damage, and may be a potential alternative for treatment of urinary stricture.