In order to determine the utility of flow cytometry for detecting metallo-beta-lactamase (MBL)-producing Pseudomonas aeruginosa, we used genotypically characterized P. aeruginosa isolates from the Molecular Epidemiology and Genetics Laboratory of the Universidad Nacional Mayor de San Marcos. A total of 29 isolates (17 MBL-producing and 12 non-MBL-producing) were analyzed with the FACSCalibur (Becton Dickinson) cell viability kit. Two treatments were used, one with meropenem and the other with meropenem-EDTA. A significant difference between MBL and non-MBL-producing P. aeruginosa was demonstrated using the fluorescence ratio in non-living cells, considering a cut-off point of >1.6. We determined a sensitivity of 94.1% and a specificity of 100%. Flow cytometry represents an alternative for the detection of MBL-producing P. aeruginosa.