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To compare objective measures of needle manipulation between students and teachers of acupuncture using motion video analysis technology, to help support instructional acupuncture education.A total of 30 teachers and 60 students participated in this study. Acupuncture needles were inserted at LI11 and motion videos were recorded for three subtypes of ‘lifting-thrusting’ manipulation: (1) ‘mild reinforcing-attenuating’; (2) ‘reinforcing’; and (3) ‘attenuating’. The videos were analysed using Simi Motion 3D software to acquire the movement parameters of four trace marks: ‘thumb tip’; ‘forefinger tip’; ‘forefinger middle joint’; and ‘forefinger base joint’. Differences between the two groups were compared using t-tests, X2 tests and/or rank-sum tests.Changes in the near-end interphalangeal joint were positively associated with a range of movement along the X axis. Motion parameters for the thumb tip, the proximal interphalangeal (PIP) joint of the forefinger and the X axis shaft swing near the end of the forefinger in the teacher group were higher than those in the student group. The teacher group featured smaller trough dispersion and smaller crest dispersion during ‘reinforcing’ and ‘attenuating’ manipulations, respectively.The ‘lifting-thrusting’ manipulation could be simplified as a fixed-axis rotation using metacarpophalangeal joints in the thumb and forefinger as the shaft centre. Teachers opened at a larger angular variation for the PIP during the lifting and thrusting processes with better spatial control. Temporal control was similar between groups and therefore appears easier to grasp. Repetitive training might be helpful for improving athletic and spatial stability during needle manipulation.