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Description: Examining the influence of culture on personality and its unbiased assessment is the main subject of cross-cultural personality research. Recent large-scale studies exploring personality differences across cultures share substantial methodological and psychometric shortcomings that render it difficult to differentiate between method and trait variance. One prominent example is the implicit assumption of cross-cultural measurement invariance in personality questionnaires. In the rare instances where measurement invariance across cultures was tested, scalar measurement invariance – which is required for unbiased mean-level comparisons of personality traits – did not hold. In this article, we present an item sampling procedure, Ant Colony Optimization, which can be used to select item sets that satisfy multiple psychometric requirements including model fit, reliability, and measurement invariance. We constructed short scales of the IPIP-NEO-300 for a group of countries that are culturally similar (USA, Australia, Canada, UK) as well as a group of countries with distinct cultures (USA, India, Singapore, Sweden). In addition to examining factor mean differences across countries, we provide recommendations for cross-cultural research in general. From a methodological perspective, we demonstrate ACO’s versatility and flexibility as an item sampling procedure to derive measurement invariant scales for cross-cultural research. The open data set used in this paper can be found at: https://osf.io/wxvth/.