The incidence of foodborne illness is higher in older adults because of their increased susceptibility; therefore, food safety practices are important. However, inadequate knowledge and negative attitudes toward food safety have been reported, which may increase use of unsafe food handling practices. Data on the actual food safety behaviors of older adults are lacking. In this study, food safety practices of older adults were observed and linked to microbiological analysis of kitchen surfaces to identify suspected routes of contamination. Older adults (≥60 years, n = 100) prepared a set meal in a model domestic kitchen sanitized according to a validated protocol to ensure minimal and consistent microbiological loads. Food safety behaviors were observed using ceiling-mounted cameras and recorded using a predetermined behavioral checklist. Surface microbiological contamination also was determined after food preparation. Overall, older adults frequently implemented unsafe food handling practices; 90% failed to implement adequate hand decontamination immediately after handling raw chicken. For older adults who used a larger number of adequate hand decontamination attempts, microbiological contamination levels in the kitchen following the food preparation session were significantly lower (P < 0.001). The novel utilization of behavioral observation in conjunction with microbiological analysis facilitated identification of potentially unsafe food handling practices as suspected routes of microbiological cross-contamination in a model domestic kitchen. Findings indicate the potential impact on domestic food safety of unsafe food handling practices used by older adult consumers. This innovative approach revealed that a large proportion of older adults implement behaviors resulting in microbiological cross-contamination that may increase the risk of foodborne illness in the home.