Numerical Calculations of Single-Cell Electroporation with an Electrolyte-Filled Capillary

Abstract

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.

Single-Cell Transfection by Electroporation Using an Electrolyte/Plasmid-Filled Capillary

“…The pEGFP-C2 plasmid has a sequence length of 4.7 kb, and our buffer is similar to the one reported, so we expect that electroosmotic flux is dominant inside the capillary. We did a simulation based on the measured capillary geometry to pursue the electric field distribution using a model similar to the one we described elsewhere 22. The simulation results reveal the electric field at the tip can be as high as ∼ 6000 kV/m, decaying rapidly in the solution outside the tip (Figure 4.…”

Electroosmotic perfusion of tissue: sampling the extracellular space and quantitative assessment of membrane-bound enzyme activity in organotypic hippocampal slice cultures

“…For our work, a concern is the effect of the electric current or field on the tissue. From earlier work on electroporation [75, 117–120] we know that an electric field of approximately 6.7×10 3 V m −1 is required to electroporate a cell of typical dimensions, approximately 20–30 µm in diameter. This is in line with the observation that a cell can be electroporated if it experiences a transmembrane potential difference of approximately 200 mV.…”

Recent Trends on Micro/Nanofluidic Single Cell Electroporation

“…The permeabilization area can be controlled with the pulse amplitude and the degree of permeabilization can be controlled with the duration of pulses, numbers of pulses, where longer pulses provide a larger perturbation area in the cell membrane [34,35]. In earlier studies of micro/nanofluidic based single cell electroporation, authors analyze cellular content and cellular properties [36][37][38][39], transfection of cells [17,[40][41][42] and inactivating cells [43][44][45] with the use of micro-channel based electroporation [46][47][48][49], micro-capillary based electroporation [50][51][52], electroporation with solid microelectrode [36,[53][54][55], membrane sandwich based microfluidic electroporation [56,57], microarray single cell electroporation [58], optofluidic based microfluidic devices [59][60][61][62][63][64][65], etc. Table 1 describes in detail micro/nanofluidic based single cell transfection, cell lysis, cell type with species, potential difference, pulse duration, etc.…”