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.
Subject(s)
Cell Culture Techniques/methods , Cell Physiological Phenomena , Electroporation/methods , Animals , CHO Cells , Cell Adhesion/physiology , Cell Line , Cricetinae , Cricetulus , Electrolytes/chemistry , Finite Element Analysis , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , PC12 Cells , Plasmids/genetics , Rats , Reproducibility of Results , Transfection/methodsABSTRACT
We here report on a concept for creating well-defined electric field gradients between the boundaries of capillary electrode (a capillary of a nonconducting material equipped with an interior metal electrode) outlets, and dielectric surfaces. By keeping a capillary electrode opening close to a boundary between a conducting solution and a nonconducting medium, a high electric field can be created close to the interface by field focusing effects. By varying the inner and outer diameters of the capillary, the span of electric field strengths and the field gradient obtained can be controlled, and by varying the slit height between the capillary rim and the surface, or the applied current, the average field strength and gradient can be varied. Field focusing effects and generation of electric field patterns were analyzed using finite element method simulations. We experimentally verified the method by electroporation of a fluorescent dye (fluorescein diphosphate) into adherent, monolayered cells (PC-12 and WSS-1) and obtained a pattern of fluorescent cells corresponding to the focused electric field.
