The majority of teachers in the early care and education (ECE) workforce face disparities in physical and psychological well-being as well as working conditions. This study examined the working conditions and physical, psychological, and professional well-being of early childhood teachers and compared the results by the child age group they care for and by teachers' level of educational attainment. Two hundred sixty-two early childhood teachers completed a brief health assessment and a questionnaire on demographic information, well-being, and working conditions. A subsample of 40 teachers were observed for the quality of care they provided to the children. A substantial number of the teachers in the study reported physical and psychological challenges and suboptimal working conditions for well-being, a largely neglected workforce issue, for this working population. Several significant differences in teachers' well-being and working conditions were found by the child age group for which they care and by teachers' level of educational attainment. In particular, teachers with higher educational levels had greater resources for work and provided higher quality of care, however, they also reported poorer physical and professional well-being than teachers with less education. Overall findings indicate a need for policies and programs that will improve working conditions and health outcomes for early childhood teachers and the children in their care. These findings suggest that ECE teachers may have unique needs across the age groups with whom they work and teacher educational levels, regarding work-related resources and conditions.