We newly developed an infrared video-oculographic system for on-line tracking of the eye position in awake and head-fixed mice, with high temporal resolution (240 Hz). The system consists of a commercially available high-speed CCD camera and an image processing software written in LabVIEW run on IBM-PC with a plug-in video grabber board. This software calculates the center and area of the pupil by fitting circular function to the pupil boundary, and allows robust and stable tracking of the eye position in small animals like mice. On-line calculation is performed to obtain reasonable circular fitting of the pupil boundary even if a part of the pupil is covered with shadows or occluded by eyelids or corneal reflections. The pupil position in the 2-D video plane is converted to the rotation angle of the eyeball by estimating its rotation center based on the anatomical eyeball model. By this recording system, it is possible to perform quantitative analysis of rapid eye movements such as saccades in mice. This will provide a powerful tool for analyzing molecular basis of oculomotor and cognitive functions by using various lines of mutant mice. (C) 2004 Elsevier Ireland Ltd and The Japan Neuroscience Society. All rights reserved.