Objectives. Religiosity and spirituality (R/S) are associated with many dimensions of human life and could contribute to one’s self-esteem; however, there is no certainty that this is also applicable to non-religious countries. Therefore, the aim of the study was to explore the association of different aspects of R/S with self-esteem in a secular environment. Participants and setting. An online sample of 464 Czech respondents aged 15 and over (mean age 30.7; SD=12.63; 27.2% men) participated in the survey. Self-esteem, religiosity, religious attendance, frequency of prayer, negative religious coping, image of God and spirituality were measured. Results. Regular prayer, spirituality (per standard deviation, SD), a low level of religious struggles (per SD) and a positive God image (per SD) were associated with higher self-esteem, with odds ratios ranging from 1.28 to 2.16 (p˂0.05 to p˂0.001). In contrast, compared to non-religious respondents, religious respondents had an approximately 60% lower chance of having a high level of self-esteem (p˂0.05). However, a combination of R and S showed that while religious/spiritual respondents did not differ significantly from non-religious respondents, religious/non-spiritual respondents had approximately 79% lower chance of having good self-esteem (p˂0.001). Study limitations. The main limitation of this study is that it did not reach a representative sample, which limits the generalizability of the findings to the whole population. This is also the first study using this kind of research approach, which, however, limits the interpretation of results. Moreover, it is a cross-sectional study, so any conclusions on causality cannot be made, and the questionnaire used only self-report measures, which could be influenced by a social desirability bias.