Optical coherence tomography (OCT) is restricted by its low scanning speed and limited resolution. High-definition raster-scanning OCT (HD-OCT) was used to evaluate changes in retinal microstructure in patients with acute central serous chorioretinopathy (CSCR) and to find new morphological features. Eighteen patients with subretinal fluid accumulation caused by acute CSCR were imaged in a cross-sectional study design. High-speed frequency-domain HD-OCT was used to image an area of 6 x 6 mm in the macular retina. Three-dimensional analyses were performed using en-face imaging and section analysis of single HD-OCT scans. Detailed information about fluid accumulation can be obtained in all compartments. Discrete changes in reflectivity are visualized within the outer nuclear or plexiform layers in > 90% of patients. Subretinal fluid appears as a dome-shaped pool of fluid and is not associated with a loss of photoreceptor layer integrity. Deposits are demarcated beneath the outer cone segments. Multiple small pigment epithelial detachments are present in > 60% of patients. High-definition OCT provides extensive information regarding precise topographic and layer-specific localization of discrete morphological changes. Along with well-known changes in the retinal microstructure, hyper-reflectivity can be imaged in the outer nuclear or plexiform layers and may represent intraretinal changes indicating the presence of subretinal pathologies or retinal maladjustment caused by the underlying pathology.