Background and Purpose-Because the blood circulation system of retina and brain are closely related to each other, we examined whether stroke is associated with localized retinal nerve fiber layer defects (RNFLDs). Methods-Patients with acute ischemic stroke as part of a hospital-based study group were compared with the participants of the population-based group Beijing Eye Study. The retina was imaged by spectral-domain optical coherence tomography for the detection of localized RNFLDs. Results-The study included 154 patients with acute ischemic stroke and 2890 subjects from the Beijing Eye Study for whom optical coherence tomographic images of the retinal nerve fiber layer and data on a previous cerebral stroke were available. In logistic regression analysis, acute stroke was significantly associated with localized RNFLDs (P<0.001; odds ratio, 6.23; 95% confidence interval, 4.17-9.30) after adjusting for age, male sex, arterial hypertension, diabetes mellitus, and higher concentration of the C-reactive protein. In a similar manner, previous stroke was associated with localized RNFLDs (P=0.04; odds ratio, 1.48; 95% confidence interval, 1.02-2.16) in multivariate analysis. In a reverse manner, presence of localized RNFLDs was associated with cerebral stroke (P<0.001; odds ratio, 3.54; 95% confidence interval, 2.68-4.67) after adjusting for age, sex, and prevalence of diabetes mellitus. Conclusions-Localized RNFLDs showed a strong association with previous or acute cerebrovascular stroke and vice versa after adjustment for other systemic and ocular factors. Localized RNFLDs that can be assessed by noninvasive optical coherence tomographic imaging may be added to the panoply of retinal morphological features of stroke.