Background: Professional autonomy is a multidimensional concept and considered a main nursing professional priority in expanding the scope of the nursing practice, so nurses require greater autonomy and participation in decision making in an organization. The current phenomenological study will offer an opportunity to improve the understanding and application of autonomy as a concept in different healthcare settings. Aim: to describe the lived experience of nursing staff about challenges and opportunities regarding their professional autonomy. Design: A phenomenological qualitative research design was used. Sample: A Purposive sample of (n=23) nursing staff was recruited for this study. Setting: General New Cairo Hospital which is affiliated to Ministry of Health. Tools: Two tools were used for data collection: Personal characteristic data sheet and semi-structured interview using digital voice recording. Results: four themes were identified as challenges were faced by nursing staff to practice professional autonomy; professional, individual, organizational and interpersonal relationship-related challenges with 34 sub-themes. Data added that professional challenges had six subthemes; individual challenges had eight subthemes, while the organizational challenges had seventeen subthemes and three subthemes are emerged under interpersonal relationship challenges. Regarding opportunities, only nine participants reported different opportunities that can enhance nurses’ autonomy such as: continuous training, available needed resources, nursing administration support unit manager and staff nurses, mutual nurse-physician respect and nurses' participation in patient care decision making. Conclusion: there are four challenges faced nursing staff to practice autonomously such as professional, individual, organizational and interpersonal relationship challenges and it was concluded that there are limited available opportunities. Recommendations: developing professional guidelines based on the lived experience of nursing staff and developing and offering training program for newly hired nurses about professional autonomy.