Objectives: To evaluate the pupillary light reflex responses in patients with unilateral strabismic and anisometropic amblyopia using dynamic pupillometry. Materials and Methods: A total of 102 eyes of 51 patients with unilateral amblyopia were included in this cross-sectional study. Of the 51 patients, 37 (72.5%) had strabismic amblyopia and 14 (27.5%) had anisometropic amblyopia. All patients underwent complete ophthalmological examination, and pupillary light reflex responses were measured using a computerized dynamic pupillometry system (MonPack One; Metrovision, France). Initial pupil diameter; the amplitude, latency, duration, and velocity of pupil contraction; and the latency, duration, and velocity of pupil dilation were recorded. Results obtained from the patients' amblyopic and normal fellow eyes were compared using paired-samples t-test and Wilcoxon signed rank test. Results: The mean age of the patients was 11.9±6.0 years. Amblyopic eyes had longer contraction latency (p=0.009), shorter contraction duration (p=0.002), and higher dilation velocity (p=0.033) compared to fellow eyes, while other parameters did not show significant differences. In subgroup analysis, eyes with strabismic amblyopia had longer contraction latency (p=0.006) and shorter contraction duration (p=0.017), while eyes with anisometropic amblyopia had shorter contraction duration (p=0.030) when compared with fellow eyes. Conclusion: In this study, the objective records obtained by dynamic pupillometry showed that pupillary light reflex responses are affected in amblyopic eyes. This finding may shed light on unclear aspects of the pathophysiology of amblyopia.