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The present study focuses on exploring the differences and relationship between well-being and experience of pastoral and psychological service of religious denomination based on religious affiliation during the first wave of the pandemic in Czechia. Our research has been focused on the investigation, comparison, and correlation between the level of well-being and pastoral and psychological service. The research sample (n = 1126) consisted of the Czech health population with age over 16 years, of which 42.4% were men (n = 478) and 57.5% were women (n = 648). From the perspective of religiosity, the study sample was divided in terms of religion into two groups—51.9% participants with religious affiliation (n = 584) and 48.1% participants without religious affiliation (n = 542). The level of well-being was identified by means of The Satisfaction with Life Scale (Diener, Emmons, Larsen, & Griffin, 1985). The level of experience with pastoral and psychological service was measured using our non-standardised questionnaire. The results confirmed the differences between the variables of well-being and positive experience with pastoral and psychological service based on religious affiliation. Moreover, we confirmed the hypothesis of a positive correlation between well-being and positive experience with pastoral and psychological service in Czechia.