Abstract Background Research on laterality in emotion suggests a dichotomy between the brain hemispheres. The present study aimed to investigate this further using a modulated startle reflex paradigm. Methods We examined the effects of left and the right ear stimulation on the modulated startle reflex (as indexed by eyeblink magnitude, measured from the right eye) employing short (2 min) film clips to elicit emotions in 16 right-handed healthy participants. The experiment consisted of two consecutive sessions on a single occasion. The acoustic startle probes were presented monaurally to one of the ears in each session, counterbalanced across order, during the viewing of film clips. Results The findings showed that eyeblink amplitude in relation to acoustic startle probes varied linearly, as expected, from pleasant through neutral to unpleasant film clips, but there was no interaction between monaural probe side and foreground valence. Conclusions Our data indicate the involvement of both hemispheres when affective states, and associated startle modulations, are produced, using materials with both audio and visual properties. From a methodological viewpoint, the robustness of film clip material including audio properties might compensate for the insufficient information reaching the ipsilateral hemisphere when using static pictures. From a theoretical viewpoint, a right ear advantage for verbal processing may account for the failure to detect the expected hemispheric difference. The verbal component of the clips would have activated the left hemisphere, possibly resulting in an increased role for the left hemisphere in both positive and negative affect generation.