The application of nanofluid in enhancing oil recovery (EOR) is eliciting considerable interest from researchers. The present study investigates the impact of a novel environmentally friendly nanofluid, composed of rhamnolipid and SiO 2 nanoparticles, on oil-water distributions in micro/nano pore spaces, suggesting the potential in EOR from low-permeability reservoirs. We conduct comprehensive experiments, including contact angle measurement, micromodel tests, and nuclear magnetic resonance (NMR) core displacement experiments, to research the synergistic effects of biosurfactant and SiO 2 nanoparticles on oil-water distribution changes. Firstly, the contact angle of a liquid droplet on rock surfaces provides a quantitative characterization of water phase spreading on pore walls. The results indicate that the nanofluid injection is advantageous in enhancing the swept volume of the water phase. Secondly, the visualization of fluid mobilization and migration in micro/nano pores is demonstrated by micromodel test. Residual oil is mobilized from stagnant zones with nanofluid injection, and subsequently recovered by waterflooding. Moreover, the micromodel observations exhibit that the nanofluid can disperse the trapped oil into small oil droplets and displace them. Finally, the NMR core displacement experiments clarify the main EOR mechanisms in microporous spaces using nanofluid. As compared to brine and rhamnolipid solution, nanofluid is more likely to motivate trapped oil, thereby increasing the swept volume of the injection fluids. Besides, the distribution changes of residual oil are also influenced by the reservoir's permeability. As the permeability of experimental cores decreases from 48.5 mD to 0.1 mD, the swept volume of nanofluid gradually expands from larger to smaller pores (from >1000 nm to 1−10 nm). The nanofluid decreases the residual oil saturation of three cores by 3.97%, 4.97%, and −1.65%, respectively, compared to the rhamnolipid-only solution. The increase in residual oil saturation observed in the core sample with 0.1 mD suggests that the re-aggregate of activated oil induces changes in oilwater distribution.