The early care and education (ECE) workforce provides care for children aged zero to five. This critical segment of the workforce experiences significant burnout and turnover rates resulting from extensive demands, including job stress and poor overall well-being. Factors associated with well-being in these settings and the resulting impacts on burnout and turnover are understudied. The purpose of this study was to investigate associations between five well-being domains and burnout and turnover outcomes among a large sample of Head Start ECE staff in the United States. An 89-item survey based off the National Institutes of Occupational Safety and Health Worker Wellbeing Questionnaire (NIOSH WellBQ) was administered to ECE staff employed in five large urban and rural Head Start agencies. The WellBQ is made up of five domains intended to measure worker well-being as a holistic construct. We utilized linear mixed-effect modeling with random intercepts to investigate associations between sociodemographic characteristics, well-being domain sum scores and burnout and turnover. After controlling for sociodemographic variables, the well-being Domain 1 (Work Evaluation and Experience) (β = -.73, p < .05) and Domain 4 (Health Status) (β = -.30, p < .05) were significantly and negatively associated with burnout; the well-being Domain 1 (Work Evaluation and Experience) (β = -.21, p < .01) was significantly and negatively associated with turnover intent. These findings suggest that multi-level well-being promotion programs could be critical to mitigate ECE teacher stress and address individual-, interpersonal-, and organizational-level predictors of overall ECE workforce well-being.