Abstract The retinal tissue is highly susceptible to inadequate oxygenation due to its high metabolic activity. Indeed, it is commonly accepted that retinal hypoxia plays a central role in the events leading to neuronal cell damage and death due to ischemia and implicated in visual impairments due to retinal vascular occlusions and diabetic retinopathy. Previous studies have investigated the use of hyperbaric oxygen treatment for retinal ischemia. However, the effectiveness of this treatment to supply adequate oxygen to the inner retina is limited and a standard of care does not exist. We address an unmet need for the development of therapeutic interventions to oxygenate the retinal tissue and prevent vision loss from an ischemic insult. We propose the concept of oxygenation of the inner retina with oxygen nanobubbles (ONBs) that can release oxygen over an extended period of time as a novel treatment modality for retinal ischemia. Our aims are to: develop a consistent formulation for the synthesis of ONBs and evaluate its physical and biochemical properties; identify the optimum time for administration of ONBs treatment to improve ischemia-induced impairments of retinal oxygen metrics and visual function; and determine the efficacy of ONBs treatment based on a series of comprehensive outcomes, namely, retinal oxygen metrics, visual function, retinal thickness, and biochemical pathways. We expect our findings will serve as strong preliminary evidence of an innovative therapeutic intervention to mitigate ischemic insult, preserve the retinal tissue from injury, and prevent vision loss.