Optical coherence tomography (OCT) provides not only morphological information but also information about layer-specific optical intensities, which may represent the underlying tissue properties. The purpose of this study is to quantitatively investigate the optical intensity of each retinal layers in central retinal artery occlusion (CRAO). Twenty-nine CRAO cases at acute phase and 33 normal controls were included. Macula-centered 3D OCT images were segmented with a fully-automated Iowa Reference Algorithm into 10 layers. Layer-specific mean intensities were determined and compared between the patient and control groups using multiple regression analysis while adjusting for age and optical intensity of the entire region. The optical intensities were higher in CRAO than in controls in layers spanning from the retinal ganglion cell layer to outer plexiform layer (standardized beta = 0.657 to 0.777, all p < 0.001), possibly due to ischemia. Optical intensities were lower at the photoreceptor, retinal pigment epithelium (RPE), and choroid layers (standardized beta = −0.412 to −0.611, all p < 0.01), possibly due to shadowing effects. Among the intraretinal layers, the inner nuclear layer was identified as the best indicator of CRAO. Our study provides in vivo information of the optical intensity changes in each retinal layer in CRAO patients.