Introduction: Non-invasive measurement of blood oxygen saturation via spectroscopic imaging has facilitated insights into the development and progression of a variety of ocular conditions, including retinal vascular occlusion, diabetic retinopathy and glaucoma. Major developments since the late 90s have been enabled by advancements in imaging technology, computational image analysis, and experimental methods. Areas covered: We review the theory of spectroscopic oximetry, the ocular blood vessels targeted for oximetry, imaging systems used for oximetry, and oximetry validation methods. Important physiological and clinical insights provided by oximetry in the eye are detailed. Expert commentary: Oximetry has revealed physiological norms and auto-regulatory effects in the retina, choroid, episcleral, and bulbar conjunctival blood vessels. Retinal oximetry has provided crucial insight into the development of diabetic retinopathy and glaucoma, and has enhanced the evaluation and treatment of retinal vessel occlusion. Commercially available retinal oximetry systems have enabled oximetry in the clinic. The development more sophisticated phantoms that resemble in vivo environments has oximetry validation in diverse oximetry applications. New insights into ocular physiology and disease are likely to be gleaned from future studies.