Traditional diagnostic testing for dermatophyte infection currently requires skin scraping for light microscopy and/or fungal culture or skin biopsy. Immunofluorescent microscopy can also be used with calcofluor stain. All of these tests can be time-consuming to perform, require a waiting period for results and are invasive. This study aimed to define the in vivo reflectance confocal microscopy (RCM) features of superficial cutaneous fungal infections and to analyse concordance with microscopic examination. Totally, 45 patients, who were diagnosed with superficial cutaneous fungal infections according to the positive result of microscopic examination, were enrolled in this study. We selected three typical lesions examined by RCM, and then recorded the results. In the patients with the tinea manus and pedis, mycelium in stratum corneum was found by the RCM in 14 of 22 patients (14/22; 63.64%). In the patients with the tinea cruris, mycelium in stratum corneum was found by the RCM in 19 of 23 patients (19/23; 82.61%). RCM seems to be useful for microscopic evaluation of mycelium features and may have a scientific value in study of superficial cutaneous fungal infections.