Objective To establish a simple reliable and accurately method of non-invasive intratracheal intubation in mice. Methods Female C57BL/ 6 mice were used. Mice were anesthetized with pentobarbital sodium, then suspended via the upper incisors using surgical thread. An epidural anesthesia catheter, 1 mL syringe, 10 μL pipet tip, were assembled as intratracheal intubation equipment. Using the non-dominant hand to feel the trachea, the dominant hand inserted the equipment into the mouth cavity to deliver the liquid. The mice were administered trypan blue or PBS. After intratracheal intubation, mice that received trypan blue were killed and the lungs were checked, while the mice that received PBS remained under observation for monitoring of survival. Results The lungs of mice that received trypan blue were dyed blue, while the stomach was not dyed. The mice that received PBS remained alive after recovering from the anesthesia. Conclusions Equipment comprising an epidural anesthesia catheter, a 1 mL syringe, a 10 μL pipet tip, and ophthalmic forceps enable simple and accurate non-invasive intratracheal intubation.