Simple SummaryEye examination is crucial for therapeutic plans and rehabilitation of birds in wildlife rehabilitation centers. However, fundus examination using classical direct or indirect ophthalmoscopy techniques can be challenging in those species. The aim of the study was to assess the use of a smartphone-based retinal imaging system in birds. Fundus examination was feasible in most bird species examined in this study. The difficulties of carrying out the examination seem to be related to the form of the globe, the color of the iris, and the quality of pupil dilation. Further investigations are necessary to confirm these findings.Ophthalmic examination is essential in the avian triage process in order to apply prompt therapeutic plans and evaluate rehabilitation potential. Fundoscopy is traditionally performed by direct or indirect ophthalmoscopy. Recent technological developments have enabled the design of a small-sized and affordable retinal imaging system to examine the fundus. We investigate the use of a smartphone-based device to realize fundus examination through a prospective cross-sectional observational study. Seventy-seven eyes of 39 birds of 15 different species were evaluated using the smartphone-based device in a rescue wildlife center. Pupil dilation was achieved prior to examination via rocuronium topical application. Assessment of fundus by the smartphone was classified as satisfactory, moderately satisfactory, and unsatisfactory. Fundus examination was also performed with a 20D, 30D, or 78D lens for comparison. Pupillary dilation was satisfactory, moderately satisfactory, or absent in 17, 32, and 28 eyes, respectively. Fundus examination with the smartphone-based device was satisfactory, moderately satisfactory, or unsatisfactory in 44, 15, and 18 eyes, respectively. The feasibility of the fundus examination was affected by the form of the globe; by the quality of pupil dilation; by the color of the iris (images could not be obtained from species with an orange, bright iris); and by the species, with owls (Strigiformes) being the easiest to observe. Based on these findings, fundus examination was feasible in most bird species examined in this study.