Pupillary light reflex (PLR) is an involuntary response where the pupil size changes with luminance. Studies have shown that PLR response was altered in children with autism spectrum disorders (ASDs) and other neurological disorders. However, PLR in infants and toddlers is still understudied. We conducted a longitudinal study to investigate PLR in children of 6-24 months using a remote pupillography device. The participants are categorized into two groups. The 'high risk' (HR) group includes children with one or more siblings diagnosed with ASDs; whereas the 'low risk' (LR) group includes children without an ASD diagnosis in the family history. The participants' PLR was measured every six months until the age of 24 months. The results indicated a significant age effect in multiple PLR parameters including resting pupil radius, minimal pupil radius, relative constriction, latency, and response time. In addition, the HR group had a significantly larger resting and minimal pupil size than the LR group. The experimental data acquired in this study revealed not only general age-related PLR changes in infants and toddlers, but also different PLRs in children with a higher risk of ASD. Autism Spectrum Disorders (ASDs) are complicated disorders that are marked by persistent deficits in social communication and interactions and by restricted, repetitive patterns of behavior, interests or activities 1. Initially chronicled 75 years ago 2 , ASDs now affect about 2.47% children and adolescents in USA alone 3. Although the etiology of ASD is still not fully understood, our understanding of this disorder has since been significantly improved owing to a large amount of physiological, psychological, and neurological studies. Evidence supports that the outcome in children with ASDs can be greatly improved by using early behavioral intervention 4,5. Unfortunately, most children do not receive an ASD diagnosis until after the age of four 6 , although early signs may appear as young as 12 months of age 7. Therefore, there is a great interest in finding effective biological markers for early screening of risk of autism and assessing responses to interventions. Pupillary light reflex (PLR) is the involuntary and nearly instantaneous pupil size change that occurs as a response to the luminous intensity of light that falls on the retina. The pupil size is controlled by the dilator and sphincter muscles innervated primarily by the sympathetic and parasympathetic branches of the autonomic nervous system (ANS), respectively 8. In 1961, Rubin observed that the pupils in 7 to 12 years old children with ASD constricted slower in responses to light adaption compared to typically developing children 9. Using a computerized pupillography system, Fan et al. discovered that pupils of children with ASD took a greater amount of time to respond to short (0.1 s) light stimuli and constricted less and more slowly than those with typical development 10. Similar atypical PLR responses were also reported in subsequent studies in children with ASD of diffe...