Urbanization is a major driver of the global loss of biodiversity; to mitigate its adverse effects, it is essential to understand what drives species' patterns of habitat use within the urban matrix. While many animal species are known to exhibit sex differences in habitat use, adaptability to the urban landscape is commonly examined at the species level, without consideration of intraspecific differences. The high energetic demands of pregnancy and lactation in female mammals can lead to sexual differences in habitat use, but little is known of how this might affect their response to urbanization. We predicted that female Pipistrellus pygmaeus would show greater selectivity of forging locations within urban woodland in comparison to males at both a local and landscape scale. In line with these predictions, we found there was a lower probability of finding females within woodlands which were poorly connected, highly cluttered, with a higher edge : interior ratio and fewer mature trees. By contrast, habitat quality and the composition of the surrounding landscape were less of a limiting factor in determining male distributions. These results indicate strong sexual differences in the habitat use of fragmented urban woodland, and this has important implications for our understanding of the adaptability of bats and mammals more generally to urbanization.