This work has developed and evaluated a microemulsion-based drilling fluid formulation with characteristics to be applied in oil wells. The microemulsion was formulated with a solution of water/glycerol, pine oil, and Tween 80, a nonionic and biodegradable surfactant. The physical and chemical properties of the drilling fluid obtained in this work were investigated through rheology and filtration analysis, solids content, aging, lubricity, toxicity, and thermal degradation. A non-toxic microemulsion-based drilling fluid oil-in-water (O/W) with high lubricity (0.07638) and thermal stability was obtained with suitable viscosity, gel strength and low fluid loss (4.0 mL), low solids content (6%), stability in a wide range of salinity conditions, and the possibility of high water content (above 85% in mass fraction). The fluid presented a pseudoplastic behavior, and statistically significant Herschel–Bulkley parameters were obtained.