In this article, a new method is introduced for estimating the motion of the heart due to respiration in gated cardiac SPECT using a rigid-body model with rotation parametrized by a unit quaternion. The method is based on minimizing the sum of squared errors between the reference and the deformed frames resulting from the usual optical flow constraint by using an optimized conjugate gradient routine. This method does not require any user-defined parameters or penalty terms, which simplifies its use in a clinical setting. Using a mathematical phantom, the method was quantitatively compared to the principal axis method, as well as an iterative method in which the rotation matrix was represented by Euler angles. The quaternion-based method was shown to be substantially more accurate and robust across a wide range of extramyocardial activity levels than the principal axis method. Compared with the Euler angle representation, the quaternion-based method resulted in similar accuracy but a significant reduction in computation times. Finally, the quaternion-based method was investigated using a respiratory-gated cardiac SPECT acquisition of a human subject. The motion-corrected image has increased sharpness and myocardial uniformity compared to the uncorrected image.