Probing living cells permeabilization dynamics by terahertz attenuated total reflection

Using attenuated total reflection (ATR) in the terahertz domain, we demonstrate non-invasive, non-staining real time measurements of cytoplasm leakage during permeabilization of live epithelial cells by saponin detergent and after electropermeabilization. The origin of the contrast observed between cells and culture medium is addressed by both experimental and theoretical approaches, and gives access to permeabilization dynamics of live cells in real time. We show that terahertz modalities are more sensitive than fluorescence microscopy which is the reference optical technique for electropermeabilization. We propose analytical models for the influx and efflux of non-permeant molecules through permeabilized cell membranes.

[1]  R. Persson,et al.  Differential permeabilization of membranes by saponin treatment of isolated rat hepatocytes. Release of secretory proteins. , 1987, The Biochemical journal.

[2]  Antoine Wojdyla,et al.  Attenuated internal reflection terahertz imaging. , 2013, Optics letters.

[3]  Peter Cresswell,et al.  Ionic contrast terahertz near-field imaging of axonal water fluxes , 2007 .

[4]  Naoshi Kondo,et al.  Characterization of Dielectric Responses of Human Cancer Cells in the Terahertz Region , 2014 .

[5]  G. Gallot,et al.  Quantitative measurement of permeabilization in living cells by terahertz attenuated total reflection , 2015, 2015 40th International Conference on Infrared, Millimeter, and Terahertz waves (IRMMW-THz).

[6]  Monitoring the molecular composition of live cells exposed to electric pulses via label-free optical methods , 2020, Scientific Reports.

[7]  I. Hapala,et al.  Breaking the barrier: methods for reversible permeabilization of cellular membranes. , 1997, Critical reviews in biotechnology.

[8]  G. Gallot,et al.  Dynamics of Cell Membrane Permeabilization by Saponins Using Terahertz Attenuated Total Reflection. , 2020, Biophysical journal.

[9]  G. Gallot,et al.  Relative Contributions of Core Protein and Solvation Shell in the Terahertz Dielectric Properties of Protein Solutions. , 2017, The journal of physical chemistry. B.

[10]  Jean-Baptiste Masson,et al.  Ionic contrast terahertz time resolved imaging of frog auricular heart muscle electrical activity , 2006 .