Early stationary phase cells of Staphylococcus aureus were inoculated into a model food, simulated milk ultrafiltrate (SMUF) and subjected to 16, 32, and 64 pulses at electric field intensities of 20, 40 and 60 kV/cm at 13 °C. In addition temperatures of 20, 25 and 30 °C were also tested with 32 pulses and an electric field of 60 kV/cm. The temperature of the SMUF increased by 1-2 ° C at the end of the 64 pulses. Cells subjected to 64 pulses at 20, 40 and 60 kV/cm were observed for ultrastructural changes using scanning and transmission electron microscopy techniques. The cell surface was rough after treatment with electric field when observed by scanning electron microscopy (SEM). The cell wall was broken, and the cytoplasmic contents were leaking out of the cell after exposure to 64 pulses at 60 kV/cm when observed by transmission electron microscopy (TEM). The breaking of the cell wall is an indication of electro-mechanical breakdown of the cell. The increase in inactivation with an increase in the electric field strength can be related to the increase in the damage to the cells. Cells subjected to 32 pulses at 60 kV/cm and 13, 20 or 25 °C were compared microscopically with the untreated control cells. Cells subjected to heat treat ment (10 min, at 66 °C) were compared with electric field-treated and untreated control cells. Although important changes were observed in the protoplast, no cell wall breakdown was observed in heat-treated cells when compared to the electric field-treated cells. This result indi cates a different mechanism of inactivation of cells with heat treatment.
Direct comparison of off-axis holography and differential phase contrast for the mapping of electric fields in semiconductors by transmission electron microscopy.
