Scanning Electroporation of Selected Areas of Adherent Cell Cultures

Abstract

We present a computer-controlled scanning electroporation method. Adherent cells are electroporated using an electrolyte-filled capillary in contact with an electrode. The capillary can be scanned over a cell culture and locally deliver both an electric field and an electroporation agent to the target area without affecting surrounding cells. The instantaneous size of the targeted area is determined by the dimensions of the capillary. The size and shape of the total electroporated area are defined by these dimensions in combination with the scanning pattern. For example, striped and serpentine patterns of electroporated cells in confluent cultures can be formed. As it is easy to switch between different electroporation agents, the method is suitable for design of cell cultures with complex composition. Finite element method simulations were used to study the spatial distributions of the electric field and the concentration of an electroporation agent, as well as the fluid dynamics related to scanning and flow of electroporation agent from the capillary. The method was validated for transfection by introduction of a 9-base-pair-long randomized oligonucleotide into PC12 cells and a pmaxGFP plasmid coding for green fluorescent protein into CHO and WSS cells.

Method for Electric Parametric Characterization and Optimization of Electroporation on a Chip

“…Equipped with a computer-controlled scanning system, the capillary electrode was scanned over a cell culture to acquire specific patterns of electroporated cells. 28 Although it is an excellent method to generate a focused electric field pattern, their method is not suitable for electroporation parameter optimization and characterization. In their system, the electric field is focused, which means that the electrical field distributes with a steep slope and the distribution is sensitive to the gap between the capillary electrode and dielectric surface.…”

Micro-/nanofluidics based cell electroporation

“…Nevertheless, there are efforts to achieve high throughput in MEP and NEP using array-type and flow-through setups. [65][66][67]71,[73][74][75][76][77][78][79][80][81][82][83][84][85] In these devices, either electric pulses or a DC field is imposed on the cell solution occupied the entire flow channel, or as solution droplets containing multiple cells. These technologies are briefly summarized here.…”

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.…”