Harmony is regarded as important for well-being in many cultures. However, (cultural) differences in the meanings and manifestations of harmony may impact the equivalence of measures of harmony in life, as well as the associations between harmony and other well-being constructs. This study aimed to investigate the factorial, convergent and divergent validity, and measurement invariance of the Harmony in Life Scale (HILS) in South African and Ghanaian samples. Confirmatory factor analysis was applied to data from three South African samples (two multicultural samples completed the HILS in English; and a Setswana-speaking sample completed the HILS in Setswana) and one Ghanaian sample (completed the HILS in English). Sample sizes ranged between n = 400 and n = 427. Good fit indices were obtained for all samples, except for the Setswana-speaking sample from South Africa. In all instances the HILS showed good internal consistency reliability and convergent and divergent validity. Full scalar invariance was supported for the two multicultural South African samples, but only partial scalar invariance when data from the Ghanaian sample were added to the analysis. The HILS shows potential for future use in all samples, except the Setswana-speaking sample. Findings emphasise the importance of considering cultural and/or contextual and linguistic differences and how these may influence the measurement of psychological constructs. Future research should qualitatively explore the meanings and manifestations of harmony in various African and other global contexts in local languages.Contribution: This study is the first to investigate the psychometric properties of the original English version of the HILS in South African and Ghanaian samples, as well as a Setswana translation of the scale. The study contributes to the understanding of harmony in life and the measurement thereof in diverse contexts, in this case specifically focused on African samples, and may, in turn, inform interventions and evaluation of interventions.