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Leaders in the healthcare setting are challenged with competing responsibilities as they seek to provide high-quality services, ensure the implementation of safety measures, and engage in workforce maintenance. Many researchers have described innovation as a strategic approach to organizational concerns and have noted a failure to implement innovative measures in healthcare. This study was an investigation of the impact of ambidexterity in healthcare leaders on innovation. The purpose of this quantitative study, guided by the ambidexterity theory of leadership for innovation, was to analyze the extent to which ambidextrous leadership characteristics of healthcare executives and chief nursing officers (CNOs) influence the innovative performance of CNOs in the healthcare setting. The research question addressed which ambidextrous leadership behaviors or combination of behaviors, including open and closed behaviors of healthcare executives and exploration and exploitation actions of CNOs in healthcare settings, explain a statistically significant portion of the variance in innovative performance of CNOs. The research design involved the administration of a cross-sectional quantitative survey to 126 CNOs from across the nation. Each participant was employed by an acute-care hospital, held an active nursing license in the state employed, and had been in the role of CNO or chief nursing executive (CNE) for over a year, reporting directly to executive leadership. A linear multiple regression analyzed the correlation between ambidextrous factors and the effect on performance innovation. Findings suggest that a combination of ambidextrous behaviors had more impact on innovation performance than each set of behaviors alone. The results of the study may be used for the development of nursing leadership and supporting efforts for overall improvement in healthcare.