Significance The involvement of the subthalamic nucleus (STN) in affective processing has been suggested with the appearance of neuropsychiatric side effects of deep brain stimulation in Parkinson’s disease (PD), but direct evidence has been lacking. In our study, we recorded single-neuron activity from the STN during affective picture presentation to PD patients intraoperatively. We discovered two spatially distinct populations of “affective” neurons responding to the emotional dimensions of the stimuli: valence (pleasantness-unpleasantness) and arousal (intensity). As previously believed, neural circuits underlying these two affective dimensions are functionally segregated. Here we observed separated emotional processing even at the single neuron level. These results extend our knowledge regarding the emotional role of the STN and the neural basis of emotions.