Electroporation devices

FIGURE 13.8 An electrical circuit diagram for a simple electroporation device. 

Transfer the sample to an ice-cold cuvette taking care to dry the outside of the cuvette. Place the cuvette in the electroporation device and deliver to the cells an electrical pulse of 1.8 kV for approximately 5 msec (use a capacitance of 10 pF and a resistance of 600 ohm). 

Electroporation efficiency depends on the parameters of electric pulses that are delivered to the treated cells using specially designed electrodes and electronic devices. In vitro experiments usually employ parallel plate types of electrodes made of inert metals like stainless steel or platinum but needle types of electrodes are also used for tissue electroporation [24,25,27,28] as well as for tumor treatment apphcations [29-32]. There are two types of electroporator devices available devices with voltage output and those with current output. However, a voltage output device seems to be preferable, which is widely used for diverse applications. 

Evans et al. [371] reported humoral and cellular immune responses elicited in response to a novel DNA epitope-based vaccine (AV-1955) delivered to rhesus macaques using the TriGrid electroporation device. AV-1955 generates long-term, potent anti-Ap antibodies and cellular immune responses specific to foreign T-helper epitopes but not to self-Ap. 

Gene Pulser (BioRad, Hercules, CA) or other electroporation device capable of... 

Compared with the traditional electroporative devices, a very low-intensity electric pulse is usually required to perform microfluidic cell electroporation. This is due to the extremely small space of the electrodes in microfluidic electroporative devices. It significantly reduces the size of the required pulse generator, the amoxmt of generated heat, and safety cautions... 

One of the main applications of microfluidic electroporation devices is cell lysis. In these devices, the mechanical (shear force) or electrical forces are applied to rapture the cell membrane and release its intercellular contents. 

Many studies have been conducted on this topic. As an example. Fig. 3 depicts the schematic diagram of the microfluidic electroporative device proposed by Wang et al. [12]. In their setup, cells start to move from sample reservoir (right-hand side) to the receiving reservoir (left-hand side). The electrodes are placed at the ends of the microchannels. In the electroporation area, geometric change and reduction of the cross-sectional area of the microchannels intensifies the applied electric field to the required electric field of electroporation. 

Lu H, Schmidt MA, Jensen MF (2005) A mierofluidic electroporation device for cell lysis. Lab Chip 5 23-29... 

Several electroporation devices were developed and are being used (Potter, 1988). In the protocols described here the devices used were the Gene Pulser from Bio-Rad (Cat. No. 165-2977) and a device constructed at EMBL (available on request). Electroporation cuvettes with 0.2-cm and 0.4-cm electrode gaps are from Bio-Rad (Cat. Nos. 165-2088 and 165-2086). 

Alternatively, electrocompetent bacteria and a suitable electroporation device are required. 

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