High repeatability of needle insertion experiments is essential to the needle-phantom interaction model validation. However, the influential factors governing the accuracy of the phantom and needle deformations have not been systematically studied. In this paper, the impact of influential factors, including phantom characteristic represented by the ratio of DMSO and thawing time (TT), needle properties represented by needle external diameter (NED) and operating factors such as needle insertion velocity (Ⅳ), insertion positions (IP) and repeated insertion times (RITs) are analyzed by orthogonal experiment design. The range calculation shows the most sensitive parameters to phantom deformations are RITs, Ⅳ and DMSO while the most sensitive parameters to needle deflection are DMSO, TT and NED. By variance analysis, the significant factors on maximum tissue deformation (MTD) are Ⅳ, followed by RITs, DMSO and IP. And NED and TT have nearly no significant impact on MTD. The significant sequence on maximum needle deflec-tion (MND) is as follows: DMSO, TT and NED. Results show that, among all impacting factors, phantom deformation is susceptible to both material properties and operative factors while the needle deflection is more susceptible to mate-rial properties of the phantom, which can help researchers in related fields to conduct experiments in a more precise manner and better understand the needle-phantom interaction mechanism.