Background: Many studies have investigated the application of micro-insulated needles with radio frequency (RF) to treat acne in humans; however, the use of a micro-insulated needle RF applicator has not yet been studied in an animal model. Objectives: The purpose of this study was to evaluate the effectiveness of a micro-insulated needle RF applicator in rabbit ear acne (REA) model. Methods: We investigated the effect of selectively destroying the sebaceous glands using a micro-insulated needle RF applicator on the formation of comedones induced by application of 50% oleic acid and intradermal injection of Propionibacterium acnes in the orifices of the external auditory canals of rabbits. The effects of the device were evaluated using regular digital photography, 3D Primos® imaging evaluation, skin Visiometer® microscopic photography and histologic analyses. Results: Use of the applicator resulted in successful selective destruction of the sebaceous glands and attenuated TNF-alpha release in an REA model. The mechanisms may involve inhibition of comedone formation, triggering of the wound healing process, and destruction of the sebaceous glands and papules. Conclusion: The use of micro-insulated needles with RF applicators provides a safe and effective method for improving the appearance of symptoms in an REA model. The current in vivo study confirms that the micro-insulated needle RF applicator is an effective alternative treatment for acne.