title: Interpulse multifrequency electrical impedance measurements during electroporation of adherent differentiated myotubes.
abstract: In this study, electrical impedance spectroscopy measurements are performed during electroporation of monolayers of differentiated myotubes. The time resolution of the system (1 spectrum/ms) enable 860 full spectra (21 frequencies from 5 kHz to 1.3 MHz) to be acquired during the time gap between consecutive pulses (interpulse) of a classical electroporation treatment (8 pulses, 100 μs, 1 Hz). 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 1200 V/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.
authors.affiliation: Electronic and Biomedical Instrumentation Group, Department of Electronic Engineering, Universitat Politècnica de Catalunya, Barcelona, Spain. Electronic address: tomas.garcia.sanchez@upc.edu.
authors.forename: Tomás
authors.initials: T
authors.lastname: García-Sánchez
authors.affiliation: CNRS, UMR8203 Vectorologie et thérapeutiques anti-cancéreuses, Institut Gustave-Roussy, Villejuif, France; Univ. Paris-Sud, UMR8203, Orsay, France.
authors.forename: Antoine
authors.initials: A
authors.lastname: Azan
authors.affiliation: CNRS, UMR8203 Vectorologie et thérapeutiques anti-cancéreuses, Institut Gustave-Roussy, Villejuif, France; Univ. Paris-Sud, UMR8203, Orsay, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Leray
authors.affiliation: Electronic and Biomedical Instrumentation Group, Department of Electronic Engineering, Universitat Politècnica de Catalunya, Barcelona, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Rosell-Ferrer
authors.affiliation: Electronic and Biomedical Instrumentation Group, Department of Electronic Engineering, Universitat Politècnica de Catalunya, Barcelona, Spain.
authors.forename: Ramon
authors.initials: R
authors.lastname: Bragós
authors.affiliation: CNRS, UMR8203 Vectorologie et thérapeutiques anti-cancéreuses, Institut Gustave-Roussy, Villejuif, France; Univ. Paris-Sud, UMR8203, Orsay, France. Electronic address: Luis.MIR@gustaveroussy.fr.
authors.forename: Lluis M
authors.initials: LM
authors.lastname: Mir
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D017097:Electric Impedance; D018274:Electroporation; D051379:Mice; D018485:Muscle Fibers, Skeletal / Q000166:cytology*
keywords: Adherent cells; Cole model; Electrical impedance spectroscopy; Electrical model; Electroporation