This study presents the Dielectrophoresis (DEP) technique for the detection of Enterobacter Aerogenes (EA) pathogen by their velocity bacteria trajectory. The DEP particle manipulation was used as non uniform AC electric field. It was capable to manipulate and separate the EA pathogen from their medium deionised (DI) water. It was important to determining the DEP force (FDEP) and the unique crossover frequency range (COFu) of EA pathogen. The FDEP and Claussius-Mossotti Factor (CMF) or polarization factor of EA were analysed and simulated by finite element method (FEM) first. The parameters of DEP experimental were fixed at 10 Volt peak to peak (Vpp) with frequency range starting from 300 kHz to 15000 kHz. The DEP microelectrode with one dropped of 0.2µL EA suspension was monitored and recorded by eyepiece camera for observed the EA movement. The DEP experimental was successfully conducted. The EA species displayed one peak for PDEP and one peak for NDEP. The highest PDEP velocity was 80 μm/s at 1100 kHz and the highest NDEP velocity was 40 μm/s at range frequency of 8000 kHz until 10000 kHz. Thus, the COFu was ranged from 1200 kHz until 1300 kHz.