Clotrimazole is an antifungal agent that has been shown to have excellent in vitro activity against most strains of Acanthamoeba. We encountered four patients who developed Acanthamoeba keratitis while wearing contact lenses that had been stored in homemade saline. Their medical treatment regimens included the use of topical 1% clotrimazole. In two patients in whom conventional therapy failed, clotrimazole was successful in controlling recurrent infection following penetrating keratoplasty. Two other patients were treated with clotrimazole as well as propamidine isethionate and neomycin sulfate-polymyxin B sulfate-gramicidin from the outset, and had an excellent response to medical therapy. In those patients who found the commercially available cream uncomfortable, a 1% clotrimazole suspension formulated in artificial tears was used and found to be well tolerated.