DC microsecond-pulsed

A comparison of atomization and ionization processes in dc, rf and microsecond pulsed discharges revealed the greatest mass loss to occur with the last type of source (two orders of magnitude greater than with the other two) [251]. 

While vesicle deformation in AC fields concerns stationary shapes, DC pulses induce short-lived shape deformations. In different studies, the pulse duration has been typically varied from several microseconds to milliseconds, while studies on cells have investigated a much wider range of pulse durations-from tens of nanoseconds to milliseconds and even seconds [80], as discussed in other chapters of this book. Various pulse profiles, unipolar or bipolar, as well as trains of pulses have been also employed (e.g., [81, 82]). Because the application of both AC flelds and DC pulses creates a transmembrane potential, vesicle deformations of similar nature are to be expected in both cases. However, the working fleld strength for DC pulses is usually higher by several orders of magnitude. Thus, the degree of deformation can be different. 

Use of a high field to activate a membrane enzyme was first reported by Witt et al. (25) in 1976. They used dc pulses of approximately 1 kV/cm and of 1-ms duration to induce ATP synthesis by the chloroplast ATPase. Following this initial work, there have been many reports on 1-kV/ cm dc field-induced ATP synthesis in different ATP synthetic systems (see the literature cited in references 13 and 14). The main conclusion from these studies is that an applied field-induced transmembrane potential can facilitate ATP release from the enzyme whether a PEF can affect enzyme turnover is not clear. Because 1-kV/ cm dc fields also cause severe Joule heating of a sample suspension, thermal effects cannot be easily avoided except when very short electric pulses (microseconds) are used. Thus, the method has limited utility for electroactivation experiments. The PEF method is, however, quite popular for the study of electroporation and electrofusion of cell membranes (see the chapter by J. Weaver in this volume), electroinsertion of membrane proteins (26), and electrotransfection of cells (27). 

The pulses may be of unidireetional eurrent (interrupted DC, monophasic), which implies that the current has a DC component. High-voltage pulsed galvanic stimulation is also used, with pulse currents up to some amperes, but pulse duration only a few microseconds. If DC effects are to be avoided (e.g., to reduce electrolytic effects or electrode metal corrosion), the current is biphasic. Faradic currents are biphasic currents of the type generated by an induction coil. If the pulses are slowly increased in amplitude, then reduced, and after a pause again increased, we have a ramp or surged current. Because many effects are current controlled, it is often better to use a constant amplitude current mode than a constant amplitude voltage mode of the stimulator output. 

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