Abstract: Polymer microspheres (PMs), such as polyacrylamide, have been widely applied for enhanced oil recovery (EOR), yet with environmental concerns. Here, we report a microfluid displacement technology containing a bio-based eco-friendly material, i.e., calcium alginate (CaAlg) microspheres for EOR. Two dominant mechanisms responsible for EOR over CaAlg fluid have been verified, including the microscopic oil displacement efficacy augmented by regulating capillary force (determined by the joint action of interfacial tension and wettability between different phases) and macroscopic sweep volume increment through profile control and mobility ratio reduction. This comprehensive effectiveness can be further impacted when the CaAlg microsphere is embellished ulteriorly by using appropriate amount of sodium dodecyl sulfonate (SDS). The core flooding and nuclear magnetic resonance (NMR) tests demonstrate that CaAlg-SDS microsphere can balance the interphase property regulation (wettability alteration and IFT reduction) and rheology properties, enabling simultaneous profile control and oil displacement. Excessive introduction of SDS will have a negative impact on rheological properties, which is not favored for EOR. Our results show that the involvement of 4-mM SDS will provide the best behavior, with an EOR rate of 34.38%. This cost-effective and environmentally-friendly bio-microsphere-based microfluidic displacement technology is expected to achieve “green” oil recovery in future oilfield exploitation.