The existing mazes are mainly used to study the learning and memory of animals. However, there is still a lack of corresponding maze and method in the aspect of the observation and test of aquatic animal robots. For this purpose, the authors have developed a three-dimensional water maze combined with bilayer multi-channel which equips with stratified lines and tick marks. This device is a rectangular structure composed of one square bottom and four rectangular side walls, and the channels of every side wall are composed of one upper channel and two lower channels. The center of the upper channels is in the vertical center line of every side wall, and the two uper channels of adjacent side walls are at 90° degrees with each other, and the two lower channels of adjacent side walls are at 45° degrees with each other. There are stratified lines and tick marks on the side wall to test the spatial location and movement trajectories of aquatic animals. The carp robot was put into the water maze for the underwater experimental detection. The success rates of left and right steering at 135, 90 and 45 degrees as well as forward motion of the carp robots (n = 10) were over 60%. This study showed that the device could be used to observe and test the motion of the carp robot.