Hanne Verswyvel

Chapter 2 │ Page 43 based on voltage and current waveform measurements, was calculated to be 1.8 mJ/pulse, as described in our previous report [32]. Therefore, the total NTP treatment energy was calculated to be 0.1, 0.3, 0.4, 0.9, 1.8, and 2.7 J, corresponding to the treatment times of 1, 3, 4, 10, 20, and 30 seconds. Table 1. Electrical & Operating Parameters of NTP Treatment Electrical Parameters Pulse voltage amplitude 30 kV Pulse rise time 1-1.5 µs Pulse width 2 µs Pulse frequency 50 Hz Operating Parameters Application distance 1 mm Energy per pulse 1.8 ± 0.5 mJ/pulse Treatment time 1, 3, 4, 10, 20, and 30 s NTP treatment energy 0.1, 0.3, 0.4, 0.9, 1.8, and 2.7 J The microsecond-pulsed DBD system was used to treat cells in 96-well plates once 75% confluence was reached. For experiments testing the e ect of ambient light, cells were taken out of the incubator and exposed to the environmental light in the cell culture hood (MSC-Advantage 1.8, ThermoFisher Scientific) for 4 hours prior to the NTP treatment (Figure 2). Cell culture medium was removed immediately before treatment and the DBD electrode was lowered into the well using a zpositioner. The electrode was fixed at 1 mm above the cells for NTP treatment, and fresh cell culture medium was added into the well right after treatment. Following treatment of all wells, the plate was incubated at 37oC and 5% CO2 until further analysis.

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