Cortical spreading depression (CSD) is a wave of neuronal and glial depolarization that propagates across the cortex at a rate of 2–5mm/min accompanied by reversible electroencephalogram (EEG) suppression, a negative shift of direct current (DC) potential, and change of optical intrinsic signals (OIS). Propagation velocity of CSD is an important parameter used to study this phenomenon. It is commonly determined in an electrophysiological way that measures the time required for a CSD wave to pass along two electrodes. Since the electrophysiology technique fails to reveal the spreading pattern of CSD, velocity calculated in this manner might be inaccurate. In this study, we combined the electrophysiological recording and OIS imaging (OISI) for detecting changes in DC potential and OIS during CSD simultaneously. An optical method based on OISI to determine the CSD velocity, which is measured by generating a series of regions of interest (ROI) perpendicular to the advancing wavefront along propagation direction of CSD at different time points and then dividing by the distance between ROIs over time, is presented. Comparison of the accuracy of the two approaches in determining the CSD velocity is made as well. The average rate of 33 CSDs is 3.52 ± 0.87mm/min by use of the optical method and 4.36 ± 1.65mm/min by use of the electrophysiological method. Because of the information about spreading pattern of CSD provided optically, the velocity determined by OISI is of smaller deviation and higher accuracy.