Modern scanning electron imaging (SEM) has become an essential tool in various fields. In order to obtain high‐quality images with minimal vibration and drift, the multi‐frame stacking technique is commonly used. However, one aspect that has been overlooked in this process is the electron beam‐induced temperature effect. This study aims to explore the impact of the temperature effect on SEM contrast for a single spherical nanoparticle, specifically a gold nanosphere on a carbon substrate. Through Monte‐Carlo simulation, we investigated how the number of frames in multi‐frame stacking mode affects two types of electron signals: secondary electrons (SE) and backscattering electrons (BSE). Our findings revealed that the SEM contrast is indeed sensitive to the number of frames used in the stacking process. It is observed that the SE contrast decreases while the BSE contrast increases compared to cases without considering temperature effects. It is also found that when using the large primary electron beam energy, the impact of temperature effect was less significant. The electron‐solid interaction theory was utilized to systematically explain the underlying mechanism. This study not only sheds light on how temperature effects can influence SEM imaging but also provides valuable insights for optimizing image acquisition techniques.