BACKGROUND: Acupuncture sensation and manipulation have been considered to be an important component of acupuncture in traditional Asian medicine. However, there has been limited research as to whether acupuncture sensation is associated with therapeutic benefit. This study investigated the relationship between acupuncture sensation and analgesic effect according to acupuncture manipulation. METHOD: Fifty-three healthy volunteers received three different forms of acupuncture in a single-blinded crossover design: superficial needling (0.3 cm), deep needling (2 cm) and needling with bi-directional rotation. The effects of acupuncture were evaluated by using the pressure pain threshold. Acupuncture sensation measurement was done in two ways. RESULTS: Both total acupuncture sensation and increase of the pressure pain threshold were maximum in needling with rotation, followed by deep needling and superficial needling. Repeated-measure analysis of variance (ANOVA) analysis was carried out to assess whether there was a significant difference; both showed significant difference (p = 0.000, 0.003). A paired sample t-test was carried out, which revealed that needling with rotation showed significant difference from both superficial needling and deep needling. Further, the correlation between the total acupuncture sensation and changes in pressure pain threshold were calculated using Pearson correlation; there was a significant correlation (p = 0.002, p = 0.013). CONCLUSION: Acupuncture sensation and pressure pain threshold increase according to the depth and rotation of acupuncture. Especially, both display significant increase with needle rotation. Further, there is a significant correlation between acupuncture needling sensation and increase in pressure pain threshold. It seems that needle rotation and acupuncture sensation play an important role in verifying the effect of acupuncture.