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Abstract BackgroundInsects have evolved complex visual systems and display an astonishing range of adaptations for diverse ecological niches. Species ofDrosophila melanogastersubgroup exhibit extensive intra- and interspecific differences in compound eye size. These differences provide an excellent opportunity to better understand variation in insect eye structure and the impact on vision. Here we further explored the difference in eye size betweenD. mauritianaand its sibling speciesD. simulans. ResultsWe confirmed thatD. mauritianahave rapidly evolved larger eyes as a result of more and wider ommatidia thanD. simulanssince they recently diverged approximately 240,000 years ago. The functional impact of eye size, and specifically ommatidia size, is often only estimated based on the rigid surface morphology of the compound eye. Therefore, we used 3D synchrotron radiation tomography to measure optical parameters in 3D, predict optical capacity, and compare the modelled vision to in vivo optomotor responses. Our optical models predicted higher contrast sensitivity forD. mauritiana, which we verified by presenting sinusoidal gratings to tethered flies in a flight arena. Similarly, we confirmed the higher spatial acuity predicted forDrosophila simulanswith smaller ommatidia and found evidence for higher temporal resolution. ConclusionsOur study demonstrates that even subtle differences in ommatidia size between closely relatedDrosophilaspecies can impact the vision of these insects. Therefore, further comparative studies of intra- and interspecific variation in eye morphology and the consequences for vision among otherDrosophilaspecies, other dipterans and other insects are needed to better understand compound eye structure–function and how the diversification of eye size, shape, and function has helped insects to adapt to the vast range of ecological niches.