Abstract: Mixing a Triton X-114 solution at its critical micelles concentration with an equivalent volume of bacteria sample solution was used as an effective method to enhance the quality of matrix-assisted laser desorption/ionization time-of-flight(MALDI-TOF) spectra and increased the amount of peaks. A series of experiments were also conducted to further validate the conclusion. A new method in the identification of bacterium using MALDI-TOF mass spectrometry can be made.