A microchannel was fabricated with glass tubes to investigate the effect of viscosity, surface tension, and flow rate on the liquid-liquid two-phase flow regime. Water and gasoil were selected as aqueous and organic working fluids, respectively. The two fluids were injected into the microchannel and created either slug or parallel profile depending on the applied conditions. The range of Reynolds and capillary numbers was chosen in such a way that neither inertia nor interfacial tension forces were negligible. Xanthan gum was used to increase viscosity and Triton X-100 (TX-100) and Sodium Dodecyl Sulfate (SDS) were used to reduce the interfacial tension. The results demonstrated that higher value of viscosity and flow rate increased interfacial area, but slug flow regime remained unchanged. The two surfactants showed different effects on the flow regime and interfacial area. Addition of TX-100 did not change the slug flow but decreased the interfacial area. In contrast, addition of SDS increased interfacial area by decreasing the slug’s length in the low concentrations and by switching from slug to parallel regime at high concentrations.