The purpose of this study was to formulate topically effective controlled release ophthalmic fluconazole liposomal formulations using the reverse-phase evaporation technique. Soya bean phosphatidylcholine (PC) and cholesterol (Ch) in specific weight ratios were used. Selected formulations were tested for their in-vivo ocular antifungal effect. These included the neutral, the positively (using stearyl amine) and the negatively (using dicetyl phosphate) charged liposomes. A reproducible model of Candida keratitis in rabbits was performed and the effects of the prepared liposomes were better than a solution of fluconazole. The order of fluconazole liposomal formulations according to the time to achieve complete healing is arranged in a descending order: negatively charged liposomes > positively charged liposomes > neutral liposomes (7:4) > neutral liposomes (5:5) > fluconazole solution. The frequency of instillation was decreased; also, the time of ulcer healing was decreased. It was concluded that the use of liposomes as a drug delivery system could contribute to the enhancement of the effect of fluconazole in the eye.