Plasma source ion implantation (PSII) with negative high voltage pulses has been applied to the sterilization process as a technique suitable for sterilization of three-dimensional work pieces. Pulsed high negative voltage (0-10 mus pulse width, 900 pulses/s, -9 to -16 kV) was applied to the electrode in this process at a gas pressure of 2-7 Pa of N-2. This process,has been found to be capable of generating glow discharge plasma around a stainless electrode, on which quartz glass samples with biological materials are placed. We found that the PSII process reduced the numbers of active Bacillus pumilus cell's using N-2 gas plasma generated by pulsed dc voltages. The number of bacteria survivors was reduced by 10(5) x with 5-10 min exposure. The state of cells on quartz glass was observed by scanning electron microscopy with and without exposure. We found that the ion energy is the most important processing parameter. The technique is demonstrated to be an effective means of low-temperature surface sterilization, with very little damage to the target. (C) 2003 American Vacuum Society.