A number of studies have shown that the presence of simple images of eyes in the environment increases prosocial behaviour in humans. However, questions remain about the robustness of the effect, its explanation and the factors promoting it. In particular, it is not yet clear whether this effect is restricted to contexts where there is a normative requirement to behave prosocially and thus where punishment is a likely consequence of failing to do so. In an 11-wk field experiment in a supermarket, we displayed either eye images or control images on charity collection buckets. There was no normative requirement to donate in this setting, and most people did not do so. However, the presence of eye images increased donations by 48% relative to control images. The effect of eye images was significantly stronger at times when the supermarket was quiet rather than busy. Results are consistent with models of the evolution of prosociality through reputation-based partner choice and have potential practical benefits for those involved in charitable fundraising.