Objective: Findings of existing functional MRI (fMRI) studies on the neural mechanisms that mediate effects of acupuncture analgesia are inconsistent. This study analyzes the effects of manual acupuncture on pain ratings and brain activation in response to experimental, electrical pain stimuli. Design: Fourteen healthy volunteers were examined by using a 1.5-T MRI scanner. The intensity of pain stimuli was adjusted to individual pain ratings on a numeric rating scale. Baseline fMRI was performed during electrical pain stimulation in a blocked design. For the second session, manual acupuncture with repeated stimulation was performed on contralateral acupoints-large intestine 4, liver 3, and stomach 36-before imaging. After imaging, subjective pain ratings and ratings of the de qi sensation were assessed. Results: Compared with baseline, volunteers showed modulated brain activity under pain conditions in the cingulate gyrus, insula, primary somatosensory cortex, and prefrontal areas after the acupuncture session. In accordance with the literature, anterior insular and prefrontal activity seemed to be correlated with acupuncture treatment. Conclusion: This study supports the existence of analgesic acupuncture effects that outlast the needling period. Pain-associated brain areas were modulated in direct response to a preceding acupuncture treatment.