Previous studies over recent years have revealed the presence of a resident bacterial population in the human skin throughout the entire body. However, the localization and composition of the bacteria within the epidermis and the skin appendages have not been fully investigated. Using differential tape stripping, cyanoacrylate skin surface biopsies and mapping of hair follicles, bacteria on the forearms of study participants were isolated, mapped, cultured and identified with respect to their origin within the epidermis and the hair follicles. Our studies showed that 85% of the bacteria were found within the first 6 corneocyte layers and roughly 25% of the cutaneous bacterial population were localized within the hair follicles. The microbial flora of the skin between individuals is subject to considerable fluctuations. Micrococcaceae represent the biggest fraction of hair-follicle-associated bacteria. The techniques developed for this study allowed us to selectively investigate the bacterial population within the hair follicles. Our results point out the role of skin appendages as potential microbial reservoirs and the need to develop new antiseptic formulations that sufficiently penetrate into the hair follicles.