Cervical cancer screening has reduced cervical cancer-related mortality by over 70% in countries that have achieved high coverage. However, there are significant geographic disparities in access to screening. In Ghana, although cervical cancer is the second most common cancer in women, there is no national-level cervical cancer screening program, and only 2 to 4% of eligible Ghanaian women have ever been screened for cervical cancer. This study used an exploratory, sequential mixed-methods approach to examine barriers and facilitators to cervical cancer screening from women and healthcare workers perspectives, guided by the Dynamic Sustainability Framework. Two convenience samples of 215 women and 17 healthcare personnel were recruited for this study. All participants were from one of three selected clinics (Ejisu Government Hospital, Kumasi South Hospital, and the Suntreso Government Hospital) in the Ashanti region of Ghana. Descriptive analyses were used to group the data by practice setting and ecological system. Statistical differences in means and proportions were used to evaluate women’s barriers to cervical cancer screening. Quantitative findings from the women’s survey informed qualitative, in-depth interviews with the healthcare workers and analyzed using an inductive thematic analysis. The median age of women and healthcare workers was 37.0 years and 38.0 years respectively. Most women (n=194, 90.2%) reported never having been screened. Women who had not been screened were more likely to have no college or university education. Ecologic factors identified were lack of knowledge about available services, distance to a clinic and requiring a spouse’s permission prior to scheduling. Practice setting barriers included long clinic wait times and culturally sensitive issue. The quantitative and qualitative data were integrated in the data collection stage, results, and subsequent discussion. These findings highlight the need for non-clinician-based culturally sensitive tool options for screening such as self-collected HPV tests to increase screening participation in Ghana.