A SPECT / X-ray CT system configured at Johns Hopkins University to image the biodistribution of radiopharmaceuticals in unrestrained, un-anesthetized mice has been constructed and tested on awake mice. The system was built by Thomas Jefferson National Accelerator Facility and Oak Ridge National Laboratory. SPECT imaging is accomplished using two gamma cameras, 10 cm × 20 cm in size based on a 2 × 4 array of Hamamatsu H8500 flat panel position sensitive photomultiplier tubes. A real-time optical tracking system utilizing three infrared cameras provides time stamped pose data of an awake mouse head during a SPECT scan. The six degrees of freedom (three translational and three rotational) pose data are used for motion correction during 3-D tomographic list-mode iterative image reconstruction. SPECT reconstruction of awake, unrestrained mice with motion compensation for head movement has been accomplished.