Interpulse multifrequency electrical impedance measurements during electroporation of adherent differentiated myotubes

• Published in: Bioelectrochemistry (Amsterdam, Netherlands) Vol. 105; pp. 123 - 135
• Main Authors: García-Sánchez, Tomás, Azan, Antoine, Leray, Isabelle, Rosell-Ferrer, Javier, Bragós, Ramon, Mir, LLuis M.
• Format: Journal Article Publication
• Language: English
• Published: Netherlands Elsevier B.V 01-10-2015
• Subjects: Adherent cells; Animals; Cell Differentiation; Cole model; Electric Impedance; Electrical impedance spectroscopy; Electrical model; Electroporation; Electroquímica; Enginyeria electrònica; Enginyeria química; Espectroscòpia d'impedància; Gene-transfer; High-voltage pulses; Impedance (Electricity); Impedance spectroscopy; Impedància (Electricitat); In-vivo; Irreversible electroporation; Mammalian-cells; Mice; Micro-electroporation; Monitoring electropermeabilization; Muscle Fibers, Skeletal - cytology; Patch-clamp; Química física; Single-cell electroporation; Tissue electroporation; Àrees temàtiques de la UPC; Adherent cells; Electroporation; Electrical impedance spectroscopy; Electrical model; Cole model
• ISSN: 1567-5394; 1878-562X
• DOI: 10.1016/j.bioelechem.2015.05.018

In this study, electrical impedance spectroscopy measurements are performed during electroporation of monolayers of differentiated myotubes. The time resolution of the system (1spectrum/ms) enable 860 full spectra (21 frequencies from 5kHz to 1.3MHz) to be acquired during the time gap between consecutive pulses (interpulse) of a classical electroporation treatment (8 pulses, 100μs, 1Hz). Additionally, the characteristics of the custom microelectrode assembly used allow the experiments to be performed directly in situ in standard 24 multi-well plates. The impedance response dynamics are studied for three different electric field intensities (400, 800 and 1200V/cm). The multifrequency information, analysed with the Cole model, reveals a short-term impedance recovery after each pulse in accordance with the fast resealing of the cell membrane, and a long-term impedance decay over the complete treatment in accordance with an accumulated effect pulse after pulse. The analysis shows differences between the lowest electric field condition and the other two, suggesting that different mechanisms that may be related with the reversibility of the process are activated. As a result of the multifrequency information, the system is able to measure simultaneously the conductivity variations due to ion diffusion during electroporation. Finally, in order to reinforce the physical interpretation of the results, a complementary electrical equivalent model is used.
•EIS measurements based on multisine excitations are shown to be a feasible method to capture fast electroporation dynamics.•Multifrequency information reveals a dual dynamic membrane resealing mechanism.•Impedance models are demonstrated to help in the study of electroporation.•EIS provide simultaneous information about membrane poration and the medium conductivity variations caused by ions diffusion.