NG-CANCAN Remote Targeting Electroporation: Impact of Burst Number Variation on Permeabilization Distribution in Confluent Cell Monolayers

Experiments were conducted using a four-electrode array with an inter-electrode distance of 10.0 mm. The study focused on optimizing the CANCAN protocol and investigating the effect of varying burst numbers on permeabilization distribution across confluent cell monolayers. The CANCAN protocols utilized canceling pulses to minimize cell damage near the electrodes while targeting cells in the center of the electrode array. Each single pulse in the protocol had a duration of 600 ns. The protocol consisted of nine packets of pulses delivered at a frequency of 0.2 MHz and was repeated 1, 2, 4, or 6 times at a frequency of 1 Hz. Cell monolayer integrity was assessed using Hoechst staining, while membrane permeability was evaluated using YoPro-1. This work was partially supported by NIH grant 1R21EY034258.