Picosecond lasers have emerged as the leading technology for tattoo removal due to their shorter pulse lengths. To clarify the features of picosecond lasers, we compared picosecond and nanosecond lasers in their ability to remove multi-colored tattoo in an animal model. We first compared a nanosecond quality-switched Nd:YAG laser with picosecond Alexandrite and quality-switched Nd:YAG lasers and then the picosecond quality-switched Nd:YAG laser with the picosecond Alexandrite laser, using a guinea pig model. The colors in the tattoos included red, orange, yellow, green, blue, and black. Guinea pigs were treated for one session with each type of laser. The clearance of pigmentation and local reactions were evaluated based on clinical photographic assessment, quantitative assessment using a colorimeter, histopathology, and electron microscopic examination before laser treatment, immediately after, and at 3 weeks after the treatment. Regardless of pulse duration, a 532-nm laser was the most effective in clearing red, orange, and yellow pigments, although the overall effect and safety was better with the picosecond 532 nm laser. A picosecond 755 nm laser demonstrated excellent efficacy in removing only green and blue pigments. a picosecond 1064 nm laser demonstrated some effects on non-black colored tattoos. In terms of safety, picosecond lasers produced less tissue injury than nanosecond lasers. Conclusively, picosecond lasers are more effective and safer than nanosecond lasers.