The benchmark test for preftontal cortex (PFC)-mediated working memory in rodents is a delayed alternation task utilizing variations of T-maze or Figure-8 maze, which requires the animals to make specific arm entry responses for reward. In this task, however, manual procedures involved in shaping target behavior, imposing delays between trials and delivering rewards can potentially influence the animal's performance on the maze. Here, we report an automated Figure-8 maze which does not necessitate experimenter-subject interaction during shaping, training or testing. This system incorporates a computer vision system for tracking, motorized gates to impose delays, and automated reward delivery. The maze is controlled by custom software that records the animal's location and activates the gates according to the animal's behavior and a control algorithm. The program performs calculations of task accuracy, tracks movement sequence through the maze, and provides other dependent variables (such as running speed, time spent in different maze locations, activity level during delay). Testing in rats indicates that the performance accuracy is inversely proportional to the delay interval, decreases with PFC lesions, and that animals anticipate timing during long delays. Thus, our automated Figure-8 maze is effective at assessing working memory and provides novel behavioral measures in rodents. (c) 2006 Elsevier B.V. All rights reserved.