Abstract: Objective To observe the histopathological features of piebaldism by confocal laser scanning microscopy （CLSM） in vivo and to investigate c-kit gene mutations in a family of piebaldism. Methods CLSM was applied to describe the histopathological features of piebaldism in a family, and DNA was extracted from the white blood cells of the family members to amplify the 21 coding exons of c-kit gene by PCR. Automated DNA sequencing was performed to determine the mutation sites. Results As shown by CLSM, almost no melanocytes were present in the basal layer of depigmented patches, otherwise, there was a focal or local aggregating of melanocytes at the basal layer and around the dermal papilla in the joint area of hypopigmented and hyperpigmented patches. In all patients, a T2362C mutation was found in the 17th exon of c-kit gene, which induced a change from TGT to CGT at codon 788, and resulted in a missense mutation of Cys788Arg at the highly conserved sites. However, such mutation was detected neither in the healthy family members nor in the normal controls. Conclusions CLSM is a new technique for real-time and non-invasive observation of histopathological features of skin, and can be used as an alternative option for the diagnosis of depigmented diseases. The Cys788Arg mutation may be the main cause of piebaldism in this family.