On the hair surface, cells are covered with a thin lipid layer (LL) covalently bonded to hair proteins. This integral hair lipid is different from sebaceus lipid. We conducted this study to examine the lipid distribution in human hair follicle. Transmission electron microscopy was performed to observe the ultrastructure of the LL. Hair follicles were cut and observed longitudinally along the hair axis. For transmission electron microscopy, new fixative (Lee's fixative: composed of OsO4 and RuO4) was designed as the conventional fixatives such as OsO4 or RuO4 alone were not appropriate for staining for hair follicle lipid. In addition, we measured the chemical composition of integral hair lipid by high-performance thin-layer chromatography. From the above experimental procedure, it was discovered that the lipid in the hair follicle was mainly distributed in hair cuticle and keratinized inner root sheath. A multitude of lamellar granule is observed in the vicinity of the above LL. The chemical composition of integral hair lipid was different from those of epidermal or sebaceous lipids. It is assumed that the LL in the hair follicle is similar to the epidermal LL playing an important role as a skin barrier in the stratum corneum. We proposed here the new terminology "hair barrier" from this point of view.