Electroporation-Induced Cell Modifications Detected with THz Time-Domain Spectroscopy

  • Stefania RomeoEmail author
  • P. Thomas Vernier
  • Olga Zeni


Electroporation (electropermeabilization) increases the electrical conductivity of biological cell membranes and lowers transport barriers for normally impermeant materials. Molecular simulations suggest that electroporation begins with the reorganization of water and lipid head group dipoles in the phospholipid bilayer interface, driven by an externally applied electric field, and the evolution of the resulting defects into water-filled, lipid pores. The interior of the electroporated membrane thus contains water, which should provide a signature for detection of the electropermeabilized state. In this feasibility study, we use THz time-domain spectroscopy, a powerful tool for investigating biomolecular systems and their interactions with water, to detect electroporation in human cells subjected to permeabilizing pulsed electric fields (PEFs). The time-domain response of electroporated human monocytes was acquired with a commercial THz, time-domain spectrometer. For each sample, frequency spectra were calculated, and the absorption coefficient and refractive index were extracted in the frequency range between 0.2 and 1.5 THz. This analysis reveals a higher absorption of THz radiation by PEF-exposed cells, with respect to sham-exposed ones, consistent with the intrusion of water into the cell through the permeabilized membrane that is presumed to be associated with electroporation.


Electroporation THz time-domain spectroscopy Human monocytes MM-6 cells Pulsed electric fields Water content 



The support and advice of Dr. Antonio Pepe (CNR-IREA, Napoli) in the analysis of THz time-domain spectroscopy data is gratefully acknowledged.


  1. 1.
    Joshi RP, Schoenbach KH Bioelectric effects of intense ultrashort pulses. Critical reviews in biomedical engineering 38, 255 (2010).CrossRefGoogle Scholar
  2. 2.
    Marty M, Sersa G, Garbay JR, Gehl J, Collins CG, Snoj M, et al. Electrochemotherapy—an easy, highly effective and safe treatment of cutaneous and subcutaneousmetastases: results of ESOPE (European Standard Operating Procedures of Electrochemotherapy) study. European journal of cancer 4, 3 (2006).CrossRefGoogle Scholar
  3. 3.
    Golberg A, Sack M, Teissie J, Pataro G, Pliquett U, Saulis G, et al. Energy-efficient biomass processing with pulsed electric fields for bioeconomy and sustainable development. Biotechnology for Biofuels 9, (2016).Google Scholar
  4. 4.
    Kotnik T, Pucihar G, Miklavcic D Induced transmembrane voltage and its correlation with electroporation-mediated molecular transport. The Journal of membrane biology 236, 3 (2010).CrossRefGoogle Scholar
  5. 5.
    Chizmadzhev YA, Abidor IG Membranes in strong electric fields. Bioelectrochemistry and bioenergetics 7, 83 (1980).CrossRefGoogle Scholar
  6. 6.
    Silve A, Guimera Brunet A, Al-Sakere B, Ivorra A, Mir LM Comparison of the effects of the repetition rate between microsecond and nanosecond pulses: electropermeabilization-induced electro-desensitization?. Biochimica et biophysica acta 1840, 2139 (2014).CrossRefGoogle Scholar
  7. 7.
    Lamberti P, Romeo S, Sannino A, Zeni L, Zeni O The role of pulse repetition rate in nsPEF-induced electroporation: a biological and numerical investigation. IEEE transactions on bio-medical engineering 62, 2234 (2015).CrossRefGoogle Scholar
  8. 8.
    Vernier PT, Levine ZA Water Bridges in Electropermeabilized Phospholipid Bilayers. Proceedings of the IEEE 101, 11 (2013).CrossRefGoogle Scholar
  9. 9.
    Hansen EL, Sozer EB, Romeo S, Frandsen SK, Vernier PT, Gehl J Dose-Dependent ATP Depletion and Cancer Cell Death following Calcium Electroporation, Relative Effect of Calcium Concentration and Electric Field Strength (vol 10, e0122973, 2015). PloS one 10, (2015).Google Scholar
  10. 10.
    Nesin OM, Pakhomova ON, Xiao S, Pakhomov AG Manipulation of cell volume and membrane pore comparison following single cell permeabilization with 60- and 600-ns electric pulses. Biochimica et biophysica acta 1808, 792 (2011).CrossRefGoogle Scholar
  11. 11.
    Romeo S, Wu YH, Levine ZA, Gundersen MA, Vernier PT Water influx and cell swelling after nanosecond electropermeabilization. Biochimica et biophysica acta 1828, 1715 (2013).CrossRefGoogle Scholar
  12. 12.
    Sozer EB, Wu YH, Romeo S, Vernier PT Nanometer-Scale Permeabilization and Osmotic Swelling Induced by 5-ns Pulsed Electric Fields. J Membrane Biol 250, 21 (2017).CrossRefGoogle Scholar
  13. 13.
    Azan A, Untereiner V, Descamps L, Merla C, Gobinet C, Breton M, et al. Comprehensive Characterization of the Interaction between Pulsed Electric Fields and Live Cells by Confocal Raman Microspectroscopy. Anal Chem 89, 10790 (2017).CrossRefGoogle Scholar
  14. 14.
    Azan A, Untereiner V, Gobinet C, Sockalingum GD, Breton M, Piot O, et al. Demonstration of the Protein Involvement in Cell Electropermeabilization using Confocal Raman Microspectroscopy. Sci Rep-Uk 7, (2017).Google Scholar
  15. 15.
    Heugen U, Schwaab G, Brundermann E, Heyden M, Yu X, Leitner DM, et al. Solute-induced retardation of water dynamics probed directly by terahertz spectroscopy. P Natl Acad Sci USA 103, 12301 (2006).CrossRefGoogle Scholar
  16. 16.
    Sun Y, Sy MY, Wang Y-XJ, Ahuja AT, Zhang Y-T, Pickwell-MacPherson E A promising diagnostic method: Terahertz pulsed imaging and spectroscopy. World Journal of Radiology 3, 55 (2011).CrossRefGoogle Scholar
  17. 17.
    Naftaly M, Miles RE Terahertz time-domain spectroscopy for material characterization. Proceedings of the Ieee 95, 1658 (2007).CrossRefGoogle Scholar
  18. 18.
    Chopra N, Yang K, Abbasi QH, Qaraqe KA, Philpott M, Alomainy A THz Time-Domain Spectroscopy of Human Skin Tissue for In-Body Nanonetworks. Ieee T Thz Sci Techn 6, 803 (2016).CrossRefGoogle Scholar
  19. 19.
    Miklavcic D, Sersa G, Brecelj E, Gehl J, Soden D, Bianchi G, et al. Electrochemotherapy: technological advancements for efficient electroporation-based treatment of internal tumors. Medical & biological engineering & computing 50, 1213 (2012).CrossRefGoogle Scholar
  20. 20.
    Chopra N, Yang K, Upton J, Abbasi QH, Qaraqe K, Philpott M, et al. Fibroblasts cell number density based human skin characterization at THz for in-body nanonetworks. Nano Commun Netw 10, 60 (2016).CrossRefGoogle Scholar
  21. 21.
    Reid CB, Reese G, Gibson AP, Wallace VP Terahertz Time-Domain Spectroscopy of Human Blood. Ieee J Biomed Health 17, 774 (2013).CrossRefGoogle Scholar
  22. 22.
    Kinosita K, Tsong TY Hemolysis of Human Erythrocytes by a Transient Electric-Field. P Natl Acad Sci USA 74, 1923 (1977).Google Scholar
  23. 23.
    Kinosita K, Tsong TY Formation and Resealing of Pores of Controlled Sizes in Human Erythrocyte-Membrane. Nature 268, 438 (1977).CrossRefGoogle Scholar
  24. 24.
    El Haddad J, Bousquet B, Canioni L, Mounaix P Review in terahertz spectral analysis. Trac-Trend Anal Chem 44, 98 (2013).CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.CNR – Institute for Electromagnetic Sensing of the Environment (IREA)NaplesItaly
  2. 2.Frank Reidy Research Center for BioelectricsOld Dominion UniversityNorfolkUSA

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