Objectives: The aim of this study was to suggest a new method to estimate the resistance by acupoint compositions by using the multi-frequency bioelectrical impedance analysis(MF-BIA) of 5 kHz, 50 kHz and 200 kHz within 2 cm of acupoints as a local segment. Moreover, we try to confirm the relation to between measured resistance at skin surface and measured resistance by various manual acupuncture needle insertion depth. Methods: Ten male subjects participated in this study. We measured the resistance at left/ right ST36 at skin surface and various manual acupuncture needle insertion depth(skin, 5 mm, 10 mm, 15 mm, 20 mm, 25 mm and 30 mm). Results: It was also observed that the all measured resistances were the highest at 5 kHz and the lowest at 200 kHz. There were significant differences at 5 kHz, 50 kHz and 200 kHz between measured resistance at skin surface and measured resistance by various manual acupuncture needle insertion depth(p<0.05). There was no significant difference in the left and right identical acupoints under the identical condition(p>0.05). Conclusions: We conclude that the measured resistance at skin surface has limitation as to reflect the information of tissue. However, the measured resistance at each frequency was changed as similar pattern by different insertion depth. Hence, we confirmed the possibility of assumption on information of tissue which was expected to locate an acupoint.