Early diagnosis of wound infections are crucial as they have been shown to increase patient morbidity and mortality. We evaluated the use of MolecuLight i:X to identify infections in acute open wounds in hand trauma. Data were collected from patients who attended the hand trauma unit over a 4 week period prior to having surgery. Wounds were inspected for clinical signs of infection and autofluorescence images were taken using the MolecuLight i:X device. Wound swabs were taken and results interpreted according to report by microbiologist. Autofluorescence images were interpreted by a clinician blinded to the microbiology results. 31 patients were included and data collected from 35 wounds. 3 wounds (8.6%) showed positive clinical signs of infection, 3 (8.6%) were positive on autofluorescence imaging and 2 (5.7%) of wound swab samples were positive for significant infection. Autofluorescence imaging correlated with clinical signs and wound swab results for 34 wounds (97.1%). In one case, the clinical assessment and autofluorescence imaging showed positive signs of infection but the wound swabs were negative. Autofluorescence imaging (MolecuLight i:X) in acute open wounds may be useful to provide real-time confirmation of bacterial infection and therefore guide management.