Interpulse multifrequency electrical impedance measurements during electroporation of adherent differentiated myotubes

García-Sánchez, Tomás; Azan, Antoine; Leray, Isabelle; Rosell, J.; Bragós, Ramon; Mir, Lluis M.

doi:10.1016/j.bioelechem.2015.05.018

Cited by 28 publications

(18 citation statements)

References 61 publications

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“…The achievement of cell EPN under strictly the same experimental condition as Raman experiments (5 000 cells/cm 2 platted overnight on CaF 2 substrate in 35 mm Petri dish, saline solution maintained at 4 °C, 8 electric pulses of 100 μs duration at a frequency of 1 Hz, magnitude of the electric field of 1 000 V/cm) was checked by fluorescence microscopy55. Yo-Pro-1, with λexcitation = 491 nm and λemission = 509 nm, (Life technologies) is a non-permeant intercalating fluorescence dye commonly used to check cell permeabilization5556.…”

Section: Methodsmentioning

confidence: 99%

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Demonstration of the Protein Involvement in Cell Electropermeabilization using Confocal Raman Microspectroscopy

Azan

Untereiner

Gobinet

et al. 2017

Sci Rep

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Confocal Raman microspectroscopy was used to study the interaction between pulsed electric fields and live cells from a molecular point of view in a non-invasive and label-free manner. Raman signatures of live human adipose-derived mesenchymal stem cells exposed or not to pulsed electric fields (8 pulses, 1 000 V/cm, 100 μs, 1 Hz) were acquired at two cellular locations (nucleus and cytoplasm) and two spectral bands (600–1 800 cm−1 and 2 800–3 100 cm−1). Vibrational modes of proteins (phenylalanine and amide I) and lipids were found to be modified by the electropermeabilization process with a statistically significant difference. The relative magnitude of four phenylalanine peaks decreased in the spectra of the pulsed group. On the contrary, the relative magnitude of the amide I band at 1658 cm−1 increased by 40% when comparing pulsed and control group. No difference was found between the control and the pulsed group in the high wavenumber spectral band. Our results reveal the modification of proteins in living cells exposed to pulsed electric fields by means of confocal Raman microspectroscopy.

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Section: Methodsmentioning

confidence: 99%

“…Yo-Pro-1, with λexcitation = 491 nm and λemission = 509 nm, (Life technologies) is a non-permeant intercalating fluorescence dye commonly used to check cell permeabilization5556. The level of Yo-Pro-1 fluorescence strongly increases when the dye reaches the nucleus.…”

Section: Methodsmentioning

confidence: 99%

Demonstration of the Protein Involvement in Cell Electropermeabilization using Confocal Raman Microspectroscopy

Azan

Untereiner

Gobinet

et al. 2017

Sci Rep

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“…Transdermal electroporation, which is also called electropermeabilization, takes place when the skin is exposed to high electric field pulses [ 36 ]. Molecules are able to cross the SC when the optimal electroporation parameters are chosen.…”

Section: Mechanisms Of Electroporationmentioning

confidence: 99%

“…The pores are said to be small (<10 nm), temporary and sparse (0.1% of surface area) [ 28 ]. Measurements of the electrical properties of the skin during pulse application have also been proposed as a useful technique for assessing pore evolution [ 36 ]. Some investigators have used AC signals to study the passive electrical properties of biological sample before and after electroporation at single frequencies [ 36 ].…”

Section: Mechanisms Of Electroporationmentioning

confidence: 99%

Perspectives on Transdermal Electroporation

Ita

2016

Pharmaceutics

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Transdermal drug delivery offers several advantages, including avoidance of erratic absorption, absence of gastric irritation, painlessness, noninvasiveness, as well as improvement in patient compliance. With this mode of drug administration, there is no pre-systemic metabolism and it is possible to increase drug bioavailability and half-life. However, only a few molecules can be delivered across the skin in therapeutic quantities. This is because of the hindrance provided by the stratum corneum. Several techniques have been developed and used over the last few decades for transdermal drug delivery enhancement. These include sonophoresis, iontophoresis, microneedles, and electroporation. Electroporation, which refers to the temporary perturbation of the skin following the application of high voltage electric pulses, has been used to increase transcutaneous flux values by several research groups. In this review, transdermal electroporation is discussed and the use of the technique for percutaneous transport of low and high molecular weight compounds described. This review also examines our current knowledge regarding the mechanisms of electroporation and safety concerns arising from the use of this transdermal drug delivery technique. Safety considerations are especially important because electroporation utilizes high voltage pulses which may have deleterious effects in some cases.

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“…Besides, the use of fluorescent dyes is unsuitable for subsequent assays of the treated cells. Recently, electrical detections of the current [ 26 , 36 ] and impedance [ 30 , 37 , 38 , 39 , 40 , 41 ] have been reported to be incorporated with the electroporation system. Compared to the traditional optical detections, electrical detections provide a label-free and instant strategy for electroporation assessment.…”

Section: Introductionmentioning

confidence: 99%

Single-Cell Electroporation with Real-Time Impedance Assessment Using a Constriction Microchannel

Luan

Zhang

et al. 2020

Micromachines

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The electroporation system can serve as a tool for the intracellular delivery of foreign cargos. However, this technique is presently limited by the inaccurate electric field applied to the single cells and lack of a real-time electroporation metrics subsystem. Here, we reported a microfluidic system for precise and rapid single-cell electroporation and simultaneous impedance monitoring in a constriction microchannel. When single cells (A549) were continuously passing through the constriction microchannel, a localized high electric field was applied on the cell membrane, which resulted in highly efficient (up to 96.6%) electroporation. During a single cell entering the constriction channel, an abrupt impedance drop was noticed and demonstrated to be correlated with the occurrence of electroporation. Besides, while the cell was moving in the constriction channel, the stabilized impedance showed the capability to quantify the electroporation extent. The correspondence of the impedance variation and electroporation was validated by the intracellular delivery of the fluorescence indicator (propidium iodide). Based on the obtained results, this system is capable of precise control of electroporation and real-time, label-free impedance assessment, providing a potential tool for intracellular delivery and other biomedical applications.

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Interpulse multifrequency electrical impedance measurements during electroporation of adherent differentiated myotubes

Cited by 28 publications

References 61 publications

Demonstration of the Protein Involvement in Cell Electropermeabilization using Confocal Raman Microspectroscopy

Demonstration of the Protein Involvement in Cell Electropermeabilization using Confocal Raman Microspectroscopy

Perspectives on Transdermal Electroporation

Single-Cell Electroporation with Real-Time Impedance Assessment Using a Constriction Microchannel

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