Gravity-driven membrane (GDM) filtration is one of the promising membrane bioreactor (MBR) technologies. It operates at a low pressure by gravity, requiring a minimal energy. Thus, it exhibits a great potential for a decentralized system, conducting household in developing and transition countries. Biofouling is a universal problem in almost all membrane filtration applications, leading to the decrease in flux or the increase in transmembrane pressure depending on different operation mode. Air scoring or regular membrane cleaning has been utilized for fouling mitigation, which requires increased energy consumption as well as complicated operations. Besides, repeating cleaning will trigger the deterioration of membranes and shorten their lifetime, elevating cost expenditures accordingly. In this way, GDM filtration stands out from conventional MBR technologies in a long-term operation with relative stable flux, which has been observed in many studies. The objective of this study was to monitor the biofilm development on a flat sheet membrane submerged in a GDM reactor with constant gravitational pressure. Morphology of biofilm layer in a fixed position was acquired by an in-situ and on-line OCT (optical coherence tomography) scanning at regular intervals for both visual investigation and structure analysis. The calculated thickness and roughness were compared to the variation of flux, fouling resistance and permeate quality, showing expected consistency. At the end of experiment, the morphology of entire membrane surface was scanned and recorded by OCT. Membrane autopsy was carried out for biofilm composition analysis by total organic carbon (TOC) and liquid chromatography with organic carbon detection (LC-OCD). In addition, biomass concentration was obtained by flow cytometer and adenosine tri-phosphate (ATP) method. The data of biofilm components indicated a homogeneous biofilm structure formed after a long-term running of the GDM system, based on the morphology observation by OCT images. The superiority of GDM in both flux maintaining and long-term operation with production of high quality effluent was demonstrated, as well as the suitability of OCT for biofouling monitoring was emphasized.