The aim of this research was to study the influence of both the emotional content and the physical characteristics of affective stimuli on the psychophysiological, behavioral and cognitive indexes of the emotional response. We selected 54 pictures from the IAPS, depicting unpleasant, neutral, and pleasant contents, and used two picture sizes as experimental conditions (120 x 90 cm and 52 x 42 cm). Sixty-one subjects were randomly assigned to each experimental condition. We recorded the startle blink reflex, skin conductance response, heart rate, free viewing time, and picture valence and arousal ratings. In line with previous research (e.g., Bradley, Codispoti, Cuthbert, and Lang, 2001), our data showed an effect of the affective content on all the measurements recorded. Importantly, effects of the size of the affective pictures on emotional responses were not found, indicating that the emotional content is more important than the formal properties of the stimuli in evoking the emotional response.