Background The diagnosis of central serous chorioretinopathy (CSC) is mainly dependent onfluorescine fundus angiography (FFA). However, the combination of optical coherence topography (OCT) with FFA offers a new approach to the research of the pathogenesis of CSC. Objective This clinical study was designed to study the combined application of the FFA and OCT for the research of the pathogenesis of central serous chorioretinopathy (CSC). Methods Forty-four eyes of 44 patients with CSC were included in this study with 36 cases of males and 8 cases of female. The patients were aged 39.3 ± 5.3 years and the visual acuity was 0. 64 ±0. 27. FFA and OCT examinations were performed in all patients and the FFA images were imported into the Topcon 3D OCT 1000 device to locate the conformity of OCT lesions with the leakages of FFA. The neuroepithelial layer thickness at the fovea and the height of the neuroepithelial layer detachment were measured using 3-D OCT. Results OCT showed serous REP detachment in 34 eyes (77.3%) and rough surfaces of RPE in 10 eyes (22. 7% ). In thirtyfour eyes with RPE detachment, the OCT lesions and FFA leakage spots conformed to the same locations in 31 eyes, but the other three eyes did not. The mean foveal neuroepithelial thickness was (138.5±19.4) μm in CSC eyes and that of normal eyes was ( 131.35±5. 01 ) μm ,showing a significant difference between them( t=0. 39 ,P＞0. 05 ). The mean height of neuroepithelial detachment was (263.3 ± 126.7 ) μm in CSC eyes. Conclusion RPE detachment occurs in CSC eyes and further induces macular neuroepithelial detachment. Leakage lesion of fluorescine corresponds to RPE detachment. CSC without RPE detachment may be related to the increase in RPE permeability. OCT can accurately measure the thickness of the macular neuroepithelial layer and the height of the neuroepithelial detachment.