In this work, we present a novel electroporator which is capable of generating single and bursts of high power (3 kV, 60 A) square wave pulses of variable duration (100 ns to 1 ms) with predefined repetition frequency (1 Hz to 3.5 MHz). The proposed synchronized crowbar implementation ensures a constant pulse rise and fall times, which are independent from the load, thus highly relevant in electroporation. The electroporator was successfully tested for the inactivation of the human pathogen Candida albicans. The device is compatible with standard commercial electroporation cuvettes.
Electroporation is a phenomenon occurring due to exposure of cells to Pulsed Electric Fields (PEF) which leads to increase of membrane permeability. Electroporation is used in medicine, biotechnology, and food processing. Recently, as an alternative to electroporation by PEF, Pulsed ElectroMagnetic Fields (PEMF) application causing similar biological effects was suggested. Since induced electric field in PEMF however is 2–3 magnitudes lower than in PEF electroporation, the membrane permeabilization mechanism remains hypothetical. We have designed pilot experiments where Saccharomyces cerevisiae and Candida lusitaniae cells were subjected to single 100–250 μs electrical pulse of 800 V with and without concomitant delivery of magnetic pulse (3, 6 and 9 T). As expected, after the PEF pulses only the number of Propidium Iodide (PI) fluorescent cells has increased, indicative of membrane permeabilization. We further show that single sub-millisecond magnetic field pulse did not cause detectable poration of yeast. Concomitant exposure of cells to pulsed electric (PEF) and magnetic field (PMF) however resulted in the increased number PI fluorescent cells and reduced viability. Our results show increased membrane permeability by PEF when combined with magnetic field pulse, which can explain electroporation at considerably lower electric field strengths induced by PEMF compared to classical electroporation.
Electroporation is an appealing way of stimulating living cells, which causes permanent or temporary nanoporosities in the structure of the biological objects. However, the technique has a disadvantage such as a requirement of contact between the electrodes and the cell medium. In this review, a methodology of contactless treatment of the biological objects using pulsed magnetic fields is proposed. The eukaryotic micro-organisms Achlya americana and Saprolegnia diclina have been used in the study and magnetic fields up to 7 T were applied, which caused effects similar to irreversible electroporation resulting in the death of the species. The proposed technique is applicable for different types of the biological cells or micro-organisms and possibly can be used in the area of cancer, antifungal treatment and other biotechnological fields.
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