Introduction: An online survey was conducted on 16 National Institute of Technical Education Certificate (NITEC) Dental Surgery Assistant (DSA) trainees in National Dental Centre Singapore to find out their perceptions and understanding of mental well-being. Methods: The 43-item survey included (i) Psychological General Well-being Index (PGWB) with 22 items based on 6 domains (anxiety, depression, positive well-being, self-control, general health, and vitality); (ii) 5 items on mental health knowledge; (iii) 4 items on lifestyle; and (iv) 12 items on perceptions of mental well-being and working in the dental clinic. Responses were collated for descriptive analysis and Cronbach’s alpha analysis was done for internal consistency for Likert scale items. Results: The average PGWB score was 61.5 (range 18-89). Fourteen trainees (75%) indicated they were feeling depressed and 31.2% of trainees felt under stress in the past month. The majority (81.3%) of trainees recognised the role of exercise in maintaining mental health and 75% of trainees were able to differentiate between sadness and depression. However, 56.3% and 87.5% of the trainees incorrectly answered that mental and psychological disorders are not preventable conditions and mental disorders are caused by a wrong way of thinking respectively. The Cronbach’s alpha on PGWB ( = 0.87) and trainees’ perception of working in the clinic ( = 0.76) revealed good internal consistency. Conclusion: The PGWB scores and survey analysis indicate there is a knowledge gap about mental health and the need to improve mental well-being in this cohort of NITEC DSA trainees. Keywords: Mental Well-being, Dental Surgery Assistant Trainee, Psychological General Well-being Index