Present study was carried out to develop and evaluate olive oil based nano-emulsion for transdermal delivery of fluconazole, a bistriazole based antifungal agent with poor water solubility and lipophilicity. Olive oil, a natural non-irritating, non-toxic proposed permeation enhancer, is known to have some antifungal activity as well. Screening of common emulsifiers like Tweens (Tween 20, tween 60, tween 80), Spans (span 60, span 80), brij 35, puronic 127, and poloxamer 188 were done based on solubility of fluconazole in these surfactants followed by their efficiency to emulsify olive oil in water. Co-emulsifiers such as glycols (polyethylene glycol 200, polyethylene glycol 400, propylene glycol), and short chain alcohols (ethanol, propanol, butanol and octanol) were also screened similarly. Tween 80 and butanol were selected as emulsifier and co-emulsifier respectively to formulate nanoemulsion by aqueous titration method. However, separation was observed after 24 hours. Therefore, span 80 was added as an auxiliary emulsifier to improve emulsification efficiency. Finally, a blend of tween 80, span 80 and butanol was optimized as emulsifier (56 % wt/wt) to emulsify 9 % wt/wt of olive oil in 33 % wt/wt water. Pseudoternary phase diagram was employed to identify and optimize the components. Optimized formulation based on phase separation and thermokinetic stability was characterized for globule size, size distribution, zeta potential, viscosity, refractive index and pH. Globule size analysis by zetasizer nano ZS was further confirmed by transmission electron microscopy. Permeation flux of fluconazole from optimized formulation through artificial skin was approximately three fold higher than the control. In conclusion, developed olive oil based nano-emulsion of fluconazole demonstrated promising solubility, permeability and stability. Keywords: Fluconazole, olive oil, nano-emulsion, transdermal permeation.