The feasibility of fenestration operation in middle ear bone with pulsed infrared laser is evaluated. Healthy male New Zealand rabbits in vivo are used in the experiment. Middle ear mastoid bone of animal model is completely exposed with conventional methods, and then a pulsed CO<sub>2</sub> laser (10.6 \mu m) and an Er:YAG laser (2.94 \mu m) are used to perform the fenestration operation. Diamond drill is also used as a control group. The total operation time and light irradiation time are recorded and the opening efficiency is assessed. The morphological changes and thermal damage around the opening window on the middle ear bone are examined. It is shown that both laser systems are suitable for the fenestration operation in middle ear bone, and this no-touch technique has a lot of benefits compared with traditional methods. The bleeding during operation has an important effect on operation time and thermal injury and needs to be controlled efficiently in further study.