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 2-4% 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's and healthcare workers' perspectives. These were further informed by the Dynamic Sustainability Framework (DSF), in particular, two domains, namely the practice setting and ecological system. 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. 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 were 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. Practice setting factors included long clinic wait times and distance to the clinic. Ecological system factors identified were population characteristics such as lack of knowledge about available services, shyness when undergoing a clinician-performed pelvic exam, and requiring a spouse's permission before scheduling. These findings highlight the need for non-clinician-based, culturally sensitive cervical cancer screening options such as self-collected HPV tests to increase screening participation in Ghana.