Needlestick injuries (NSI) pose a significant health hazard among healthcare personnel (HCP). The aim of our prospective observational study was to evaluate the psychological impact of NSI and assess measures to prevent NSI. The target group was the medical staff and students of Frankfurt University Hospital who had experienced a NSI (n = 370) during the 12-month study period. Data were retrieved from accident insurance reports, occupational follow-up examinations and a standardized anonymous questionnaire sent to the affected HCP. Analysis of the completed questionnaires (232/370) revealed that stress (48.3 %) and tiredness (36.6 %) were common factors contributing to the NSI and that >80 % of the respondents were concerned about the consequences of the NSI. Higher levels of anxiety were reported when the patient was known to have a chronic virus infection. Stressful working conditions, lack of adequate protective medical/technical equipment and poor work routines were suggested as factors contributing to NSI.