Various methodologies have been utilized in hand- hygiene (HH) research to measure the quality and compliance rates of hand washing. Some notable examples are direct observation, self-report, image quantification of fluorescence, microbial sampling, automated systems, and electronically assisted devices. While direct observation is considered the gold standard of HH monitoring systems, its methodological limitations (e.g., high staffing demands, participant reactivity, and undersampling) have yet to be overcome. As a result, there is renewed interest in developing technologies or methods of assessment that are cost-effective, accurate, and not intrusive. This article provides a brief review of HH monitoring systems while presenting a less resource-intensive methodology utilizing image analysis of fluorescence to assess hand washing. Results indicate that the proposed HH protocol could be used to replace human visual analysis of fluorescence, as well as provide a less resource-intensive option to assess HH under controlled conditions. Future implications and the need for additional research, such as cross-validating the results in a real-world clinical setting, are discussed.