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Improved micro-impedance spectroscopy to determine cell barrier properties

Md. Mehadi Hasan Sohag
Md. Mehadi Hasan Sohag
, 
Olivier Nicoud
Olivier Nicoud
, 
Racha Amine
Racha Amine
, 
Abir Khalil-Mgharbel
Abir Khalil-Mgharbel
, 
Jean-Pierre Alcaraz
Jean-Pierre Alcaraz
, 
Isabelle Vilgrain
Isabelle Vilgrain
 und 
Donald K Martin
Donald K Martin
   | 19. Juli 2020
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Article Category: Research Article
Online veröffentlicht: 19. Juli 2020
Seitenbereich: 150 - 155
DOI: https://doi.org/10.2478/ebtj-2020-0017
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© 2020 Md. Mehadi Hasan Sohag, Olivier Nicoud, Racha Amine, Abir Khalil-Mgharbel, Jean-Pierre Alcaraz, Isabelle Vilgrain, Donald K Martin, published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Figure 1

The electrical impedance spectroscopy (EIS) system. (A) The assembly of the microscope slide for the T10 electrode chip with its pattern of gold electrodes, and the microfluidic cartridge. Note that the slide for the T10 electrode is transparent and the gold tracks are sufficiently thin to be transparent. The red circle shows the entry and exit ports for the microfluidic channel that encloses the measurement electrode. (B) Cross-section of one microfluidic channel and ports. The arrow labelled as the entry of cells and media is via the small entry port of panel A. The cells migrate onto the gold measurement electrode (during 24 hours). The space between the electrodes is completely filled with media. The total capacity of the microfluidic chamber is 400μl. The media is changed by aspiration from the larger port of panel A. The gold measuring electrode is marked M1 and the gold reference electrode is marked M2. (C) The assembled microfluidic cartridge is inserted into the Tethapod reader and the EIS measurements are achieved using the 2 electrodes M1 and M2.
The electrical impedance spectroscopy (EIS) system. (A) The assembly of the microscope slide for the T10 electrode chip with its pattern of gold electrodes, and the microfluidic cartridge. Note that the slide for the T10 electrode is transparent and the gold tracks are sufficiently thin to be transparent. The red circle shows the entry and exit ports for the microfluidic channel that encloses the measurement electrode. (B) Cross-section of one microfluidic channel and ports. The arrow labelled as the entry of cells and media is via the small entry port of panel A. The cells migrate onto the gold measurement electrode (during 24 hours). The space between the electrodes is completely filled with media. The total capacity of the microfluidic chamber is 400μl. The media is changed by aspiration from the larger port of panel A. The gold measuring electrode is marked M1 and the gold reference electrode is marked M2. (C) The assembled microfluidic cartridge is inserted into the Tethapod reader and the EIS measurements are achieved using the 2 electrodes M1 and M2.

Figure 2

(A) EIS microfluidic cartridge seeded with the normal renal proximal tubule epithelial cells (RPTEC) in the Proxup medium adapted for their growth in culture. (B) Phase-contrast image of normal renal proximal tubule epithelial cells (RPTEC) after growing for 3 days on the electrodes of the EIS microfluidic cartridge. Note that these gold electrodes were thin enough to allow light to pass through for these images. (C) EIS microfluidic cartridge seeded with the renal cell carcinoma tumor cells (786-O) in RPMI medium supplemented with fetal bovine serum (10%). This is a different medium to that of the RPTEC cells, hence the difference in colour. (D) Phase-contrast image of renal cell carcinoma tumor cells (786-O) after growing for 3 days on the electrodes of the EIS microfluidic cartridge. Note that the growth of these tumour cells is less organized than the RPTEC normal cells.
(A) EIS microfluidic cartridge seeded with the normal renal proximal tubule epithelial cells (RPTEC) in the Proxup medium adapted for their growth in culture. (B) Phase-contrast image of normal renal proximal tubule epithelial cells (RPTEC) after growing for 3 days on the electrodes of the EIS microfluidic cartridge. Note that these gold electrodes were thin enough to allow light to pass through for these images. (C) EIS microfluidic cartridge seeded with the renal cell carcinoma tumor cells (786-O) in RPMI medium supplemented with fetal bovine serum (10%). This is a different medium to that of the RPTEC cells, hence the difference in colour. (D) Phase-contrast image of renal cell carcinoma tumor cells (786-O) after growing for 3 days on the electrodes of the EIS microfluidic cartridge. Note that the growth of these tumour cells is less organized than the RPTEC normal cells.

Figure 3

(A) The equivalent circuit used to model the EIS measurements. The paracellular resistance is represented by Rn and the capacitance of the cell monolayer is represented by Cn. The other components take into account the contributions of the electrodes (CPEm, Rm) and the media solution (Re, Cc). (B) Application the model to analyse the EIS measurement on RPTEC cells at day 1. The Bode Plot is shown on the left for the phase (■) and the impedance (●). The Nyquist plot is shown on the right. (C) Application the model to analyse the EIS measurement on 786-O cells at day 1. The Bode Plot is shown on the left for the phase (◻) and the impedance (○). The Nyquist plot is shown on the right.
(A) The equivalent circuit used to model the EIS measurements. The paracellular resistance is represented by Rn and the capacitance of the cell monolayer is represented by Cn. The other components take into account the contributions of the electrodes (CPEm, Rm) and the media solution (Re, Cc). (B) Application the model to analyse the EIS measurement on RPTEC cells at day 1. The Bode Plot is shown on the left for the phase (■) and the impedance (●). The Nyquist plot is shown on the right. (C) Application the model to analyse the EIS measurement on 786-O cells at day 1. The Bode Plot is shown on the left for the phase (◻) and the impedance (○). The Nyquist plot is shown on the right.

Figure 4

Resistance and capacitance of the cell monolayers obtained from modelling the EIS measurements. The times for measurements are the hours elapsed since the seeding of the cells, over 3 days. (A) Resistance of the cell monolayers taking into account the area of the measurement electrode (2.1 mm2). The RPTEC cells are represented by the solid bars and the 786-O cells by the hatched bars. The error bars indicate standard error of the mean. (B) Capacitancee of the cell monolayers taking into account the area of the measurement electrode (2.1 mm2). The RPTEC cells are represented by the solid bars and the 786-O cells by the hatched bars. The error bars indicate standard error of the mean.
Resistance and capacitance of the cell monolayers obtained from modelling the EIS measurements. The times for measurements are the hours elapsed since the seeding of the cells, over 3 days. (A) Resistance of the cell monolayers taking into account the area of the measurement electrode (2.1 mm2). The RPTEC cells are represented by the solid bars and the 786-O cells by the hatched bars. The error bars indicate standard error of the mean. (B) Capacitancee of the cell monolayers taking into account the area of the measurement electrode (2.1 mm2). The RPTEC cells are represented by the solid bars and the 786-O cells by the hatched bars. The error bars indicate standard error of the mean.
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