Objective To summarize the experience on tracheal intubation techniques for laboratory animals. Methods To properly choose appliance of tracheal intubation, implement appropriate approach of tracheal intubation, and prevent tracheal tube syndrome. Results The success rate of tracheal intubation reached to 100%. Conclusions A well understanding of anatomy of the larynx and trachea is the premise of a successful tracheal intubation. The technique used by us is easy to operate with a high success rate, allow to perform during large open-operation in laboratory animals, and deserves practical application.