Single-cell electroporation was performed using electrolyte-filled capillaries on fluorescently labeled A549 cells. Cells were exposed to brief pulses (50-300 ms) at various cell-capillary tip distances. Cell viability and electroporation success were measured. In order to understand the variability in single-cell electroporation, logistic regression was used to determine whether the probabilities of cell survival and electroporation depend on experimental conditions and cell properties. Both experimental conditions and cell properties (size and shape) have a significant effect on the outcome. Finite element simulations were used to compare bulk electroporation to single-cell electroporation in terms of cell size and shape. Cells are more readily permeabilized and are more likely to survive if they are large and hemispherical as opposed to small and ellipsoidal with a high aspect ratio. The dependence of the maximum transmembrane potential across the cell membrane on cell size is much weaker than it is for bulk electroporation. Observed survival probabilities are related to the calculated fraction of the cell's surface area that is electroporated. Observed success of electroporation is related to the maximum transmembrane potential achieved.

An electric field is focused on one cell in single-cell electroporation. This enables selective electroporation treatment of the targeted cell without affecting its neighbors. While factors that lead to membrane permeation are the same as in bulk electroporation, quantitative description of the single-cell experiments is more complicated. This is due to the fact that the potential distribution cannot be solved analytically. We present single-cell electroporation with an electrolyte-filled capillary modeled with a finite element method. Potential is calculated in the capillary, the solution surrounding the cell, and the cell. The model enables calculation of the transmembrane potential and the fraction of the cell membrane that is above the critical electroporation potential. Electroporation at several cell-to-tip distances of human lung carcinoma cells (A549) stained with ThioGlo-1 demonstrated membrane permeation at distances shorter than approximately 7.0 microm. This agrees well with the model's prediction that a critical transmembrane potential of 250 mV is achieved when the capillary is approximately 6.5 microm or closer to the cell. Simulations predict that at short cell-to-tip distances, the transmembrane potential increases significantly while the total area of the cell above the critical potential increases only moderately.