In a treatment-analogue experiment, extinction of fear of spiders was investigated in a group of spider-anxious students. Two groups were created: in the single extinction group the extinction trials consisted of repeated presentations of a videotaped spider in one specific location of a house, whereas in the multiple extinction group the trials consisted of videotapes of the same spider in three different locations of a house. Also a control group was included that was exposed to videotapes of the location but without the spider. As reflected in skin conductance responses and self-report data, fear of spiders was significantly reduced in the two extinction groups compared to the control group. Moreover, when the extinction groups were confronted with the videotape of the spider in a new location, the single extinction group did not show generalisation of extinction, whereas the multiple extinction group did. These results corroborate the existing evidence for context dependence of extinction of fear and provide new evidence that the use of multiple contexts during extinction might improve the generalisability of extinction in humans. Implications for exposure therapy are discussed.