Nursing is a traditional female occupation; however, the high itinerant rate of the nursing staff has been an important issue on nursing practice all the time. The study adopted a cross-sectional correlation design with the cognitive view of the profession in order to explore the ‘career self-efficacy’ and ‘career barrier’ of female nurses and to investigate the relationships between these three variables by a structured questionnaire. The research participants were female nurses from two teaching hospitals in Chiayi and Pingtung Counties. A total of 516 responded questionnaires were collected (67% retrieved). The collected data was analyzed by using the SPSS for Windows with descriptive statistics, one-way ANOVA, Scheffe’s method, and Pearson’s product moment correlation analysis. The major research findings included: First, the ‘career self-efficacy’ of the female nursing staff was considered moderate to high degree, among which the ‘intensity of confidence’ obtained the highest score(s); ‘career barriers’ fell into moderate degree, among which ‘the pressure of multiple roles’ received the highest rating, and the ‘gender discrimination’ had the lowest score. Second, personal backgrounds such as age, marital status, service seniority, professional duty, working department, specialized certificates count, and in shifts or not, all had significant influence on the career self-efficacy and career barrier. Third, except for the ‘perceived difficulty’ in career self-efficacy and ‘gender discrimination’ in career barrier, the career self-efficacy had negative correlation with the career barrier, suggesting that the higher career self-efficacy the nurses had, the lower career barrier they perceived. According to the findings, the researcher suggested implementation of formal promotion mechanism and regular shift schedule to reduce career barriers in nursing profession.