Efforts to improve the well-being of healthcare professionals include mindfulness-based interventions (MBIs). To understand the value of such initiatives, we conducted a systematic review and meta-analysis of empirical studies pertaining to the use of MBIs with healthcare professionals. Databases were reviewed from the start of records to January 2016 (PROSPERO registration number: CRD42016032899). Eligibility criteria included empirical analyses of well-being outcomes acquired in relation to MBIs. Forty-one papers met the eligibility criteria, consisting of a total of 2101 participants. Studies were examined for two broad classes of well-being outcomes: (a) “negative” mental health measures such as anxiety, depression, and stress; (b) “positive” indices of well-being, such as life satisfaction, together with outcomes associated with well-being, such as emotional intelligence. MBIs were generally associated with positive outcomes in relation to most measures (albeit with moderate effect sizes), and mindfulness does appear to improve the well-being of healthcare professionals. However, the quality of the studies was inconsistent, so further research is needed, particularly high-quality randomised control trials.