AIMS: This study aims to investigate how different wound microenvironmental factors (temperature, pH, and osmotic pressure) influence the autofluorescence of Staphylococcus aureus ( METHODS: We measured the average fluorescence intensity of RESULTS: Bacterial fluorescence intensity increased as the temperature ranged from 25°C to 43°C, with corresponding upregulation of CONCLUSIONS: Our findings indicate that temperature, pH, and osmotic pressure significantly affect the autofluorescence of