We developed an improved method for suspending and fixing rats with rat clothes in an acupuncture experiment. This protocol aims to provide detailed descriptions of its procedural steps, including important precautions, offering a reference for researchers conducting fundamental experiments in the field of acupuncture and moxibustion. In previously reported studies, researchers have often used anesthesia fixation, high platform fixation, binding fixation, or various other self-made fixators while performing acupuncture or electroacupuncture operations on rats.

Given the side effects of anesthesia on the body and the real clinical conditions, fixation in an awakened stage offers the better alternative. However, in the non-anesthetized stage, the mentioned fixations also possess limitations such as restricted acupoint exposure, requirement of multiple labels, non-adjustable fixator size, or forced posture. This method utilizes the rat's preference for burrowing to create rat jackets that fit their body shape.

It offers several advantages over other techniques. It is cost-effective, user-friendly, manpower-and space-saving, accommodates rats of varying weights, and can also be used for needle operation on most acupoints. In subsequent studies, we will focus on investigating the impact of the restraints and suspensions of rat clothing on physiological indicators and behavioral aspects related to emotional cognition in rats.

This will enable us to quantitatively assess its potential influence on research and refine the fixation method.