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Description: Emotion differentiation (ED) — the tendency to experience one’s emotions with specificity — is a well-established predictor of adaptive responses to daily life stress. Yet, there is little research testing the role of ED in self-reported and physiological responses to an acute stressor. In the current study, we investigate the effects of negative emotion differentiation (NED) and positive emotion differentiation (PED) on participants’ self-reported emotions and cardiac-mediated sympathetic nervous system reactivity (i.e., pre-ejection period) in response to a stressful task. Healthy young adults enrolled in a two-session study. At an initial session, participants completed a modified experience sampling procedure (i.e., the Day Reconstruction Method). At session 2, 195 completed the Trier Social Stress Test while cardiac impedance was acquired throughout. Linear regressions demonstrated that higher NED, but not PED, was associated with experiencing less intense self-reported negative, high arousal emotions (e.g., irritated, panicky) during the stressor (β =  − .15, p < .05) although people with higher NED also exhibited greater sympathetic reactivity (β = .16, p < .05). In exploratory analyses, we tested whether the effect of NED on self-reported stress was mediated by the tendency to make internally focus (or self-focused) attributions about performance on the task but did not find a significant indirect effect (p = .085). These results both complement prior work and provide a more complex picture of the role of NED in adaptive responses to stressful life events, suggesting that people with higher NED may experience their emotions as more manageable regardless of their level of physiological arousal.