Can Millimeter Waves Generate Electroporation?

Analysis of the field distributions in a single biological cell under electromagnetic wave is given. With Debye approximation, the dielectric relaxation of each part of the cell, including the extracellular and cellular media, the cell membrane and the nuclear membrane, was taken into account. Making use of some typical parameters for a cell, the voltage across nuclear and cytoplasma membranes under electromagnetic waves are calculated up to millimeter wave frequency range. The calculated result indicates that it is unlikely to generate electroporation by present available millimeter wave sources.     

Electric-field-induced rupture of polymer-stabilized oil films

Water-in-oil emulsions stabilized by polymeric surfactants are robust, but the reasons for their stability are poorly understood. We studied oil films stabilized by a comb–graft copolymer having a poly(siloxane) backbone and poly(ethylene oxide)/poly (propylene oxide) and C₁₆ grafts (Abil EM-90) with a total number-average molecular weight of 62,000. Electric fields imposed in the aqueous phases on either side of the oil films were used to induce rapid rupture, and the response of the film was monitored using optical interference and electrical conductance measurements. Film thickness values ranged between 30 and 50 nm and rupture at field strength values between 2 × 10⁷ and 5 × 10⁷ V/m. Unexpectedly, in some cases, stable pores were formed and the films became electrically conductive. Often the pores persisted for more than 20 min after the voltage had been removed. Since the current was independent of film area, very few pores are involved in conduction. This behavior is similar to that found in lipid films; however, the persistence time is greater for polymer-stabilized films. Because the films are thick, it is possible that pores are formed by multimolecular self-assembly as with pore-forming proteins. Polymer purification also influenced film stability. Received: 4 February 1999 Accepted: 21 May 1999     

Gene delivery to cells on a miniaturized multiwell plate for high-throughput gene function analysis

The transfectional microarray is a promising tool to conduct high-throughput analysis of gene function. In this study, a miniaturized multiwell plate was prepared by applying a silicone rubber sheet with holes on a gold electrode. The combination of electroporation and a miniaturized multiwell plate successfully achieved spatially controlled gene delivery with absence of cross-contamination between genes. Furthermore, we found that gene delivery efficiency was not dependent on conditions such as plasmid loading, electric field strength, and pulse duration.     

Electric field modulation in tissue electroporation with electrolytic and non-electrolytic additives

Electroporation, cell membrane permeabilization with short electrical field pulses, is used in tissue for in vivo gene therapy, drug therapy and minimally invasive tissue ablation. For the electroporation to be successful, the electrical field that develops during the application of the pulses needs to be precisely controlled. In this study we investigate the use of electrolytic and non-electrolytic gels to generate the precise electrical fields required for controlled electroporation, in heterogeneous and irregular tissues, in vivo. Finite element computer simulations are used to illustrate various applications, such as the treatment of irregularly shaped organs and interior cavities. The feasibility of the concept is demonstrated experimentally in vivo with a rat liver subjected to irreversible electroporation.     

Real time electroporation control for accurate and safe in vivo non-viral gene therapy

In vivo cell electroporation is the basis of DNA electrotransfer, an efficient method for non-viral gene therapy using naked DNA. The electric pulses have two roles, to permeabilize the target cell plasma membrane and to transport the DNA towards or across the permeabilized membrane by electrophoresis. For efficient electrotransfer, reversible undamaging target cell permeabilization is mandatory. We report the possibility to monitor in vivo cell electroporation during pulse delivery, and to adjust the electric field strength on real time, within a few microseconds after the beginning of the pulse, to ensure efficacy and safety of the procedure. A control algorithm was elaborated, implemented in a prototype device and tested in luciferase gene electrotransfer to mice muscles. Controlled pulses resulted in protection of the tissue and high levels of luciferase in gene transfer experiments where uncorrected excessive applied voltages lead to intense muscle damage and consecutive loss of luciferase gene expression.     

In vivo electrical impedance measurements during and after electroporation of rat liver

Electroporation is used for in vivo gene therapy, drug therapy and minimally invasive tissue ablation. Applying electrical pulses across cells can have a variety of outcomes; from no effect to reversible electroporation to irreversible electroporation. Recently, it has been proposed that measuring the passive electrical properties of electroporated tissues could provide real time feedback on the outcome of the treatment. Here we describe the results from the impedance characterization (single dispersion Cole model) for up to 30 min of the electroporation process in in vivo rat livers (n=8). The electroporation sequence consisted of 8 pulses of 100 micros with a period of 100 ms. Half of the animals were subjected to field magnitudes considered to have reversible effects (R group, E=450 V/cm) whereas for the other half irreversible field amplitudes were applied (I group, E=1500 V/cm). As expected, there was an immediate increase of conductivity (R group Deltasigma/sigma(t=0)=9+/-3%; I group Deltasigma/sigma(t=0)=43+/-1%). However, the overall long term pattern of change in conductivity after electroporation is complex and different between reversible and irreversible groups. This suggests the superposition of different phenomena which together affect the electrical properties.     

Gene transfer and protein release of fission yeast by application of a high voltage electric pulse

A high voltage electric pulse can be applied to induce the uptake of DNA into cells and the release of protein from cells. In transformation procedures, electroporation is widely used since the technique is simple, rapid, reproducible, and highly efficient. In extraction of protein, on the other hand, electroextraction has many advantages over other conventional extractions. We have developed a highly efficient method for the electroporation of fission yeast. In particular, application of a high voltage electric pulse to fission yeast improves the cellular uptake and release of macromolecules controlled by both osmotic conditions and electric field strength.

用荧光葡聚糖研究大麦细胞电融合

利用阴离子表面活性物质荧光葡聚糖 (F DX)研究了表面活性物质对大麦细胞电融合的影响 .结果表明 ,F DX可抑制电融合过程 .对放置过大麦细胞原生质体的F DX溶液 ,在荧光显微镜下可观察到其膜表面的荧光圈 ,证明F DX在膜上的吸附 .添加F DX可增加原生质体的电泳速度 ,说明吸附后原生质体表面负电荷增多 .由于相互间静电斥力的增强 ,使细胞的电融合率下降 .此外 ,还利用荧光显微技术研究了细胞电生孔现象 .观察到经电脉冲后溶液中的F DX可进入原生质体内部 ,间接证明了细胞电生孔的存在 .     

Delivering quantum dots into cells: strategies,progress and remaining issues

The use of semiconductor quantum dots (QDs) in biological sensing and labeling continues to grow with each year. Current and projected applications include use as fluorescent labels for cellular labeling, intracellular sensors, deep-tissue and tumor imaging agents, sensitizers for photodynamic therapy, and more recently interest has been sparked in using them as vectors for studying nanoparticle-mediated drug delivery. Many of these applications will ultimately require the QDs to undergo targeted intracellular delivery, not only to specific cells, but also to a variety of subcellular compartments and organelles. It is apparent that this issue will be critical in determining the efficacy of using QDs, and indeed a variety of other nanoparticles, for these types of applications. In this review, we provide an overview of the current methods for delivering QDs into cells. Methods that are covered include facilitated techniques such as those that utilize specific peptide sequences or polymer delivery reagents and active methods such as electroporation and microinjection. We critically examine the benefits and liabilities of each strategy and illustrate them with selected examples from the literature. Several important related issues such as QD size and surface coating, methods for QD biofunctionalization, cellular physiology and toxicity are also discussed. Finally, we conclude by providing a perspective of how this field can be expected to develop in the future.