BACKGROUND: Anisocoria (unequal pupil size) has been defined using cut points ranging from greater than 0.3 mm to greater than 2.0 mm for absolute difference in pupil size. This study explored different pupil diameter cut points for assessing anisocoria as measured by quantitative pupillometry before and after light stimulus. METHODS: An exploratory descriptive study of international registry data was performed. The first observations in patients with paired left and right quantitative pupillometry measurements were included. Measurements of pupil size before and after stimulus with a fixed light source were used to calculate anisocoria. RESULTS: The sample included 5769 patients (mean [SD] age, 57.5 [17.6] years; female sex, 2558 patients [51.5%]; White race, 3669 patients [75.5%]). Anisocoria defined as pupil size difference of greater than 0.5 mm was present in 1624 patients (28.2%) before light stimulus; 645 of these patients (39.7%) also had anisocoria after light stimulus (P < .001). Anisocoria defined as pupil size difference of greater than 2.0 mm was present in 79 patients (1.4%) before light stimulus; 42 of these patients (53.2%) also had anisocoria after light stimulus (P < .001). DISCUSSION: The finding of anisocoria significantly differed before and after light stimulus and according to the cut point used. At most cut points, fewer than half of the patients who had anisocoria before light stimulus also had anisocoria after light stimulus. CONCLUSION: The profound difference in the number of patients adjudicated as having anisocoria using different cut points reinforces the need to develop a universal definition for anisocoria.