Animals are exposed to different visual stimuli that influence how they perceive and interact with their environment. Visual information such as shape and colour can help the animal detect, discriminate and make appropriate behavioural decisions for mate selection, communication, camouflage, and foraging. In all major vertebrate groups, it has been shown that certain species can discriminate and prefer certain colours and that colours may increase the response to a stimulus. However, since colour is often studied together with other potentially confounding factors, it is still unclear what role colour plays in the perception of and attention to biologically relevant and familiar stimuli versus novel stimuli. In this study, we test whether colour influences the response that an animal has to an object, and if this influence – if any – changes depending on the object shown to the animal. We therefore assessed the response of three gecko species Correlophus ciliatus, Eublepharis macularius, and Phelsuma laticauda to familiar and novel objects presented as colour or grayscale images. Our results indicate that colour did not change the animal’s response to any of the objects. Specifically, we found that while all species responded more often to the novel than to the familiar images, colour did not influence the response. We also found that the duration of interaction with images was significantly longer for the diurnal species, P. laticauda, than for the two nocturnal species, but this was again independent from colouration. Finally, no differences among sexes were observed within or across species. Our results indicate that geckos discriminate between 2D images of different content independent of colouration, suggesting that colouration may not increase detectability or intensity of the response. These results are essential for uncovering which visual stimuli produce a response in animals