Background <p>Today, scales are needed that define the cultural leadership characteristics of nurse managers and its dimensions.</p> Objective <p>The aim of this study is to develop a valid and reliable assessment tool to determine the cultural leadership levels of nurse managers based on the evaluation of nurses.</p> Methods <p>This methodological study included 50 nurses for the pilot study, 355 nurses for the principal component analysis, 258 nurses for the confirmatory factor analysis, and 50 nurses for the re-test. The data were collected using the Cultural Leadership Scale in Nursing (CLS-N) and the Value-Based Leadership Scale (VBLS).</p> Results <p>The content validity index of the draft CLS-N was 0.77. According to the principal component analysis, the scale was formed with 23 items and four subscales, and the subscales accounted for 77.07% of the total variance of the scale. The fit indices of the CFA had a good or acceptable level for this construct. The Cronbach’s Alpha values for the overall scale and its subscales ranged from 0.863 to 0.952, while the item-total correlation values ranged from 0.571 to 0.743. The re-test indicated that the scale had temporal stability and the correlation values related to similar scale validity were high and significant.</p> Conclusion <p>The Cultural Leadership Scale in Nursing is a valid and reliable tool to assess the cultural leadership levels of nurse managers.</p> Clinical trial number <p>Not applicable.</p>