Abstract Background Nurses experience various health‐related issues due to the nature of their work. Aim This study aimed to examine the effect of a mindfulness‐based intervention on stress overload, depression and mindfulness among nurses. A secondary objective was to examine the role of the setting (i.e., in and out of the hospital as a natural setting) on mindfulness‐based intervention effectiveness. Methods A randomized controlled trial with three groups' pretest–posttest design was used in this study. A total of 195 nurses were recruited—65 intervention‐inside hospital, 60 intervention‐outside hospital, and 63 control. The mindfulness‐based intervention was delivered by a certified mindfulness practitioner over 4 weeks in Jordan. Data were collected using a demographics questionnaire, the Stress Overload Scale Short, the Center for Epidemiologic Studies Depression Scale‐Revised, and the Five Facet Mindfulness Questionnaire. Results Multivariate analysis showed a statistically significant effect of the intervention on the linear combination of the dependent variables V = 0.44, F [6, 368] = 17.56, p < .001. Follow‐up analyses showed that a mindfulness‐based intervention significantly decreased stress overload, depression, and increased mindfulness levels among nurses in intervention groups compared with the control group. In addition, conducting a mindfulness‐based intervention outside the hospital has a better effect on mindfulness levels than the inside hospital. Linking Evidence to Action The effect of a mindfulness‐based intervention on stress overload, depression, and mindfulness should be considered when planning for nurses' well‐being and the quality of care provided. Nurse managers are encouraged to use the study's findings to promote nurses' well‐being.