Accurate segmentation of neurosensory retinal detachment (NRD) associated subretinal fluid in spectral domain optical coherence tomography (SD-OCT) is vital for the assessment of central serous chorioretinopathy (CSC). A novel two-stage segmentation algorithm was proposed, guided by Enface fundus imaging. In the first stage, Enface fundus image was segmented using thickness map prior to detecting the fluid-associated abnormalities with diffuse boundaries. In the second stage, the locations of the abnormalities were used to restrict the spatial extent of the fluid region, and a fuzzy level set method with a spatial smoothness constraint was applied to subretinal fluid segmentation in the SD-OCT scans. Experimental results from 31 retinal SD-OCT volumes with CSC demonstrate that our method can achieve a true positive volume fraction (TPVF), false positive volume fraction (FPVF), and positive predicative value (PPV) of 94.3%, 0.97%, and 93.6%, respectively, for NRD regions. Our approach can also discriminate NRD-associated subretinal fluid from subretinal pigment epithelium fluid associated with pigment epithelial detachment with a TPVF, FPVF, and PPV of 93.8%, 0.40%, and 90.5%, respectively. We report a fully automatic method for the segmentation of subretinal fluid. Our method shows the potential to improve clinical therapy for CSC.