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An improved optical landmark-based pose measurement and tracking system has been developed to provide 3D animal pose data for a single photon emission computed tomography (SPECT) imaging system for awake, unanesthetized, unrestrained laboratory animals. The six degree of freedom animal position and orientation measurement data are time synchronized with the SPECT list mode data to provide for motion correction after the scan and before reconstruction. The tracking system employs infrared (IR) markers placed on the animal's head along with synchronized, strobed IR LEDs to illuminate the reflectors and freeze motion while minimizing reflections. A new design trinocular stereo image acquisition system using IEEE 1394 CMOS cameras acquires images of the animal with markers contained within a transparent enclosure. The trinocular configuration provides improved accuracy, range of motion, and robustness over the binocular stereo previously used. Enhanced software detects obstructions, automatically segments the markers, rejects reflections, performs marker correspondence, and calculates the 3D pose of the animal's head using image data from three cameras. The new hardware design provides more compact camera positioning with enhanced animal viewing through the 360 degree SPECT scan. This system has been implemented on a commercial gantry and tested using live mice and has been shown to be more reliable with higher accuracy than the previous system. Experimental results showing the improved motion tracking results are given.