Ambivalence, the activation of both positive and negative thoughts and feelings regarding a single attitude object, plays a role in many domains in people's lives. For instance, people can be ambivalent about societal topics, health, politics, family, and even their partners. Recently, mouse tracking has been introduced as a novel and innovative way to examine ambivalence. Although initial findings showed that mouse tracking can indeed track ambivalence in designs comparing ambivalence to positivity or negativity (i.e., univalence), it is hitherto unclear whether mouse tracking can also differentiate between ambivalence and neutrality. This is important because a) ambivalence and neutrality have distinctly different patterns of motivational activation, and b) if mouse trajectories are the same for ambivalence and neutrality, mouse tracking might not track ambivalence, but rather difficulty or uncertainty. In this paper, we examine whether neutrality can be distinguished from ambivalence in mouse tracking paradigms. Two-hundred-fifty participants evaluated neutral, ambivalent, and univalent stimuli as either positive or negative, while their mouse movements were recorded. After this, they rated the same stimuli on different self-report measures of ambivalence. First, our findings show that mouse tracking can distinguish ambivalence from univalence, replicating previous findings. More importantly, we find distinctly different patterns for ambivalent vs. neutral stimuli, demonstrating that mouse tracking can also distinguish ambivalence from neutrality. Our findings further establish mouse tracking as an innovative tool that helps researchers further their understanding of ambivalence.