Background Tinea unguium is a dermatophyte infection of the nails. Itraconazole is a broad-spectrum antifungal drug used to treat this disease. It works by interfering with the synthesis of ergosterol in the cell membrane, thus causing changes that can be seen under scanning electron microscopy (SEM). AimMethodsTo observe under SEM the in vivo ultrastructural changes of tinea unguium caused by Trichophyton rubrum before and after itraconazole (ICZ) therapy, and to explore the in vivo antifungal effect of ICZ. Ten outpatients diagnosed by mycology and SEM as having with tinea unguium caused by T. rubrum were recruited. They received ICZ pulse therapy (200 mg twice/day for 1 week, stop for 3 weeks, repeated for 3-4 months). Clinical change, mycology and SEM results were obtained after therapy. ResultsConclusionAfter ICZ therapy, 3 of the 10 cases were positive by both mycology and SEM; the latter showed completely dry, shrivelled, curved and folded hyphae with a relatively smooth surface or local destruction. Several hyphae remained intact with only some local bumps or crimps. The other cases were negative by SEM, but two were positive by mycology. In vivo ultrastructural changes of hyphae of T. rubrum in tinea unguium after ICZ treatment could be seen under SEM, demonstrating that ICZ fungistasis was effective and that SEM is a good tool for evaluating the effectiveness of ICZ in vivo. To our knowledge, this study is the first to show by SEM ICZ permeates the affected nail plate via the nail bed capillaries.