Background: Patient safety is an important aspect for quality of healthcare. Despite significant advances in medical knowledge, there are still many adverse events affecting patient safety. Therefore, the aim of this study was to assess patient safety culture and identify associated factors among healthcare professionals at public hospitals in Bahir Dar City, Northwest Ethiopia. Objective: The main objective of this study was to assess Patient safety culture and associated factors among healthcare professionals working at public hospitals in Bahir Dar City, Northwest Ethiopia, 2021. Methods: Cross sectional study design triangulated with qualitative approaches was conducted from November 10 to December 10, 2021. Simple random sampling technique was used to select 420 study participants by using lottery method. Purposive sampling technique was used for qualitative study. A standardized hospital survey on patient safety culture tool was used for data collection. Bi-variable and multivariable linear regression analyses were performed using SPSS version 23. Content analysis was used for qualitative study. Significance level was obtained at p-value <0.05 and 95%CI. Results: The overall positive patient safety culture score was 47.6% (95% CI: 42.7%, 52.5%). Increasing in age (β=1.961, CI: 1.154,2.767; p=0.000), taking patient safety training (β=0.168, CI: 0.040, 0.297; p=0.010), working in pediatric ward (β=0.236, CI: 0.099, 0.370; p=0.001), and resources availability (β=0.346, CI: 0.220, 0.473; p=0.000) were factors significantly associated with patient safety culture .The in-depth interview revealed that infrastructure, communication barrier, lack of management support, lack of good governance, healthcare professionals knowledge, skill & attitude, and patient involvement during course of treatment were identified as factors affecting patient safety culture. Conclusions and Recommendations: The overall positive patient safety culture score was low. Age of participants, taking patient safety training, working in pediatric ward, and resource availability were significantly associated with patient safety culture. Hospital managers have to avail necessary materials, work on capacity building and create good communication system. Federal Ministry of Health must develop good governance system, allocate budget for patient safety initiatives and ensuring they are adequately resourced. Keywords: Patient Safety Culture, Public Hospitals, Healthcare Professionals