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Objective: Many refugees experience prolonged separation from family members, which research suggests has adverse effects on mental health and post-displacement outcomes in refugee populations. We examine mental health differences in refugees separated and not separated from their families, and key post-migration factors and cultural mechanisms that may underlie this impact. Methods: A sample of 1085 refugees resettled in Australia, of which 23.3% were separated from all of their immediate family, took part in an online battery of survey measures indexing pre- and post-migration refugee experiences, mental health symptoms, disability and individualistic/collectivistic self-identity. Family separation was used as a predictor of mental health outcomes in a series of linear regressions, and the separated and non-separated groups were compared in multigroup path analysis models to examine group-specific indirect effects. Results: The separated group reported greater exposure to pre-migration potentially traumatic events and higher levels of post-migration living difficulties compared to the non-separated group. Family separation predicted higher post-traumatic stress and depression symptoms, but not disability, after controlling for potentially traumatic event exposure, age and sex. Path analyses revealed distinct indirect effects for separated and non-separated groups. Principally, higher collectivistic self-identity was associated with elevated post-traumatic stress, depression and disability symptoms via social-related post-migration living difficulties such as isolation and loneliness in the separated group; whereas collectivism was linked with increased depression symptoms via economic-related post-migration living difficulties in the non-separated group. Conclusion: These findings indicate that family separation powerfully influences mental health outcomes, but that its effect may be mediated by the type of post-migration stress experienced in the settlement environment and culturally bound differences in how the sense of self is interconnected with family. [ABSTRACT FROM AUTHOR]