The purpose of this study was to quantitatively assess the pupillary light reflex (PLR) in normal and anesthetized dogs using a pupillometer. Eleven dogs (20 eyes) of various breeds were included. PLRs were measured with a handheld pupillometer in dim light before and during anesthesia. Anesthesia was conducted with atropine, xylazine and ketamine. Parameters of pupillometry included neurological pupil index (NPi), pupil size, percent of change (%CH), latency (LAT), constriction velocity (CV), maximum constriction velocity (MCV) and dilation velocity (DV). NPi,%CH, CV and MCV were significantly decreased during anesthesia compared with the pre-anesthesia data. The results suggest that atropine-xylazine-ketamine combination anesthesia depresses the PLR. Additionally, this study demonstrates the feasibility of the use of a pupillometer in dogs.