A system and method is provided for calibrating and normalizing eye data of one or more subjects prior to and/or during emotional testing of the subjects. In particular, initially performing one or more calibration or normalization operations prior to an emotional test of a subject may result in accurate evaluations of emotional responses based on measurements of eye data. Additionally, further calibration or normalization performed during the emotional test may be used to refine the initial calibration or normalization, further increasing the accuracy of the evaluated emotional responses.