Background: Kindergarten education in Hong Kong serves children aged 32–68 months. However, there is no extant scale that measures kindergarten teachers’ perceived work environment, an important influence on their well-being. Aims: To develop a new instrument, the Teachers’ Perceived Work Environment (TPWE) scale, and to assess whether kindergarten teachers with higher TPWE ratings had higher scores for job satisfaction, self-esteem and mental health. Methods: A 25-item rating scale was developed and used with a sample of in-service kindergarten teachers. Their perceived work environment was represented by five factors (ergonomics, staffing, teaching space, work hours and social space). These teachers also completed three well-being inventories: the Job Satisfaction Survey, the Rosenberg Self-Esteem Inventory and the General Health Questionnaire-12. In a second stage, a new sample of in-service kindergarten teachers was used to cross-validate the findings from the earlier assessment. Results: In the first sample of 141 teachers and the second of 125, social space, staffing and work hours were associated with job satisfaction, while ergonomics was a significant negative predictor of mental health complaints. Conclusions: The TPWE exhibited satisfactory reliability and validity. Some factors were differentially associated with specific types of well-being. The results may inform future studies of the working conditions of kindergarten teachers.[Copyright © 2015 The Author.]