Midlife women frequently experience stress and menopausal symptoms. Mindfulness is thought to mitigate stress by avoiding emotional reactivity and ruminative thinking. We sought to assess the association of mindfulness and stress on menopausal symptoms among midlife women. In this cross-sectional study, women aged 40-65 years completed questionnaires, including the Menopause Rating Scale (MRS), the Perceived Stress Scale-4 (PSS-4), and the Mindfulness Attention Awareness Scale (MAAS). Linear regression was used to assess the impact of mindfulness and stress on menopausal symptoms with use of univariate and multivariable analyses, adjusting for patient characteristics. In this cohort of 1744 midlife women, higher mindfulness (MAAS) and lower stress (PSS-4) scores correlated independently with lower menopausal symptom (MRS) scores. On multivariable analysis, a significant interaction effect was observed between the MAAS and PSS-4 on the MRS, such that with higher PSS-4 scores, the magnitude of association between the MAAS and lower MRS scores was larger. Among midlife women, higher mindfulness and lower stress correlated with lower menopausal symptom scores independently. Among women experiencing more stress, the magnitude of association between mindfulness and lower menopausal symptom scores was greater, largely driven by psychological subdomain scores. Mindfulness may mitigate menopausal symptoms among midlife women.