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Background Well-being is a key aspect for healthy ageing and there is an established association between physical activity and well-being in ageing adults. Despite the recent interest in physical activity as part of a 24-h continuum also including sedentary behaviour and sleep, there is a lack of studies examining the link between these 24-h behaviours and well-being in older adults. Therefore, the aim of this study was twofold: (1) to describe 24-h behaviours and their associations with mental well-being in community dwelling adults aged 55+ years; and (2) to examine the theoretical changes in mental well-being if time were reallocated from one behaviour to another. Methods This was a cross-sectional study (n = 410). Daily time spent in sedentary behaviour, light and moderate-vigorous physical activity, and sleep was assessed using wrist-worn accelerometers during 6 days. Mental well-being was assessed using the 14-item Warwick-Edinburgh Mental Well-being Scale (WEMWBS). The association of 24-h behaviours and well-being was examined using crude and adjusted linear regression models with compositional data analysis procedures (aim 1). Associations between reallocations of five-minute intervals from five to 60 min between these behaviours and well-being were modelled using compositional isotemporal substitutions (aim 2). Results Mean age (SD) was 71.3 (6.3) years and 71% were female. In 24-h, participants spent 5.66 h asleep, 13.88 h sedentary, 2.58 h in light intensity and 1,89 h in moderate-vigorous intensity physical activity. There were no statistically significant associations between 24-h behaviours and mental well-being in fully adjusted models (aim 1). Reallocations up to 30 min were not significantly associated with changes in well-being (aim 2). There were some statistically significant theoretical changes of up to 2 points on the WEMWBS with substitutions larger than 30 min. Discussion 24-h behaviours and time reallocations between behaviours were not associated with better or worse well-being in community dwelling adults aged 55+ years. Considering well-being as a key aspect for healthy ageing, future public health research should continue to examine physical activity (both light and moderate-to-vigorous), sedentary behaviour, as well as sleep as a part of the 24-h continuum and its associations with mental health outcomes in older adults. Trial registration This trial was registered at ClinicalTrials.gov on 3th July, 2018 (Trial registration NCT03576209)