Probabilistic diffusion tractography was used to provide the first direct evidence for a subcortical pathway from the retina to the amygdala, via the superior colliculus and pulvinar, that transmits visual stimuli signaling threat. A bias to orient toward threat was measured in a temporal order judgement saccade decision task, under monocular viewing, in a group of 19 healthy participants who also underwent diffusion weighted MR imaging. On each trial of the behavioural task a picture depicting threat was presented in one visual field and a competing non-threatening stimulus in the other. The onset interval between the two pictures was randomly varied and participants made a saccade toward the stimulus that they judged to have appeared first. The bias to orient toward threat was stronger when the threatening stimulus was in the temporal visual hemifield, suggesting that afferents via the retinotectal tract contributed to the bias. Probabalistic tractography was used to virtually dissect connections between the superior colliculus and the amygdala traversing the pulvinar. Individual differences in microstructure (fractional anisotropy) of the streamline predicted the magnitude of the bias to orient toward threat, providing supporting evidence for a functional role of the subcortical SC-amygdala pathway in processing threat in healthy humans.