The novel coronavirus disease (COVID-19) pandemic has led to rising death tolls and stressed healthcare systems, resulting in an unprecedented psychological stress on healthcare workers worldwide. However, the majority of studies only accounted for frontline healthcare workers with direct patient exposure. This study aims to look at the psychological impact of COVID-19 in a specific, vulnerable and yet hidden group of healthcare workers, namely laboratory healthcare workers who are at high risk exposure to SARS-CoV-2 virus from handling infected patients' blood samples, in addition to a marked increase in workload. A multicentre study was conducted in Singapore via online questionnaire looking at psychological and physical impact of COVID-19 on laboratory healthcare workers. The Generalized Anxiety Disorder 7-item (GAD-7) scale, Zung Self-Rating Depression Scale (SDS) and Numeric rating scale on fear (NRS) were validated scores used in this study. Data analysis was performed using SPSS statistical software version 23 (IBM Corp). A total of 122 staffs participated and more than half of the cohort experienced mild to severe fear, anxiety and depression. Increase in depression score was also found to be associated with increased physical exhaustion (OR = 6.1, 95% CI 1.4-29.1, p = .02), loss of appetite (OR = 2.7, 95% CI 1.2-6.0, p = .02), poor sleep quality (OR = 7.5, 95% CI 2.9-19.4, p = .005), and the use of sedative (OR = 3.9, 95% CI 1.1-13.5, p = .03). Hence, it is imperative that prompt action needs to be taken to address the psychological needs of this vulnerable group of healthcare workers as the pandemic continues.