Application of atmospheric plasma could be used for wound healing, skin rejuvenation, and wrinkle treatment. The authors explored the feasibility of atmospheric microplasma irradiation (AMI) for enhancement of percutaneous absorption of drugs as an alternative to hypodermic needles. Pig skin was used as a biological sample exposed to AMI and analyzed by attenuated total reflection-Fourier transform infrared spectroscopy. A tape-stripping test (an evaluation method for skin-barrier performance) was also conducted to compare with AMI. Transepidermal water loss was also measured and compared with and without AMI. Results showed that surface modification of the stratum corneum (outermost skin layer) was observed upon AMI. Small pores on sample skin were observed with plasma jet irradiation due to the collision of charged particles. Percutaneous absorption was confirmed without damage upon microplasma irradiation. Our data suggested that dye pathways through skin samples could be related to the dynamic behavior of intercellular lipid bilayers, suggesting that AMI could enhance percutaneous absorption.