The paper aims to research the influences of testing device and method conditions on the puncture resistance of film and analyze factors such as material and dimensional accuracy of the testing device and conditions of test. The size and tip structure of puncture needles of different batches and materials at home and abroad were compared by microscope, and the puncture force of uniform and stable films were tested in turn. Compared with puncture needles which diameter and radius deviations were less than ±0.05 mm, the puncture force of puncture needles 1#, 4#, 5#, 6#, 7# fluctuated obviously. The needle tips with poor arc had large standard deviation and poor repeatability. Compared with the puncture needles of alloy steel, the puncture force of puncture needles with high-speed steel had better parallelism, force value was not increasing and the needle did not bend. The parallelism of puncture force which using upper and lower fixed rings with different diameters was poor. By using a puncture needle with good arc, non-deformable and accurate needle size, and together with consistent upper and lower ring diameters and reasonable fixed ring for testing, the film puncture resistance can be reflected accurately.