Electrical impedance spectroscopy measurements using a four-electrode configuration improve on-line monitoring of cell concentration in adherent animal cell cultures

Sarró, E.; Lecina, Martí; Fontova, A.; Solá, Carles Miralles; Gòdia, Francesc; Cairó, Jordi J.; Bragós, Ramon

doi:10.1016/j.bios.2011.10.028

Cited by 42 publications

(25 citation statements)

References 23 publications

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“…Transendothelial electrical resistance (TEER) measurements were performed using our custom-built multi-frequency impedance analyzer based on an AD5933 chip (Analog Devices, Nashua, NH, USA) utilizing a four-probe approach [35]. The largest changes in impedance as a function of BBB maturation were observed at 15 kHz, and all TEER measurements used in this study to evaluate the effect of LPS on BBB function are reported at that frequency.…”

Section: Methodsmentioning

confidence: 99%

Metabolic consequences of inflammatory disruption of the blood-brain barrier in an organ-on-chip model of the human neurovascular unit

Brown

Codreanu

Shi

et al. 2016

J Neuroinflammation

151

128

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BackgroundUnderstanding blood-brain barrier responses to inflammatory stimulation (such as lipopolysaccharide mimicking a systemic infection or a cytokine cocktail that could be the result of local or systemic inflammation) is essential to understanding the effect of inflammatory stimulation on the brain. It is through the filter of the blood-brain barrier that the brain responds to outside influences, and the blood-brain barrier is a critical point of failure in neuroinflammation. It is important to note that this interaction is not a static response, but one that evolves over time. While current models have provided invaluable information regarding the interaction between cytokine stimulation, the blood-brain barrier, and the brain, these approaches—whether in vivo or in vitro—have often been only snapshots of this complex web of interactions.MethodsWe utilize new advances in microfluidics, organs-on-chips, and metabolomics to examine the complex relationship of inflammation and its effects on blood-brain barrier function ex vivo and the metabolic consequences of these responses and repair mechanisms. In this study, we pair a novel dual-chamber, organ-on-chip microfluidic device, the NeuroVascular Unit, with small-volume cytokine detection and mass spectrometry analysis to investigate how the blood-brain barrier responds to two different but overlapping drivers of neuroinflammation, lipopolysaccharide and a cytokine cocktail of IL-1β, TNF-α, and MCP1,2.ResultsIn this study, we show that (1) during initial exposure to lipopolysaccharide, the blood-brain barrier is compromised as expected, with increased diffusion and reduced presence of tight junctions, but that over time, the barrier is capable of at least partial recovery; (2) a cytokine cocktail also contributes to a loss of barrier function; (3) from this time-dependent cytokine activation, metabolic signature profiles can be obtained for both the brain and vascular sides of the blood-brain barrier model; and (4) collectively, we can use metabolite analysis to identify critical pathways in inflammatory response.ConclusionsTaken together, these findings present new data that allow us to study the initial effects of inflammatory stimulation on blood-brain barrier disruption, cytokine activation, and metabolic pathway changes that drive the response and recovery of the barrier during continued inflammatory exposure.Electronic supplementary materialThe online version of this article (doi:10.1186/s12974-016-0760-y) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Metabolic consequences of inflammatory disruption of the blood-brain barrier in an organ-on-chip model of the human neurovascular unit

Brown

Codreanu

Shi

et al. 2016

J Neuroinflammation

151

128

View full text Add to dashboard Cite

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“…Each measurement included an average of 3 sweeps and required a total acquisition time of about 1 min. The technical details regarding the performance of the front-end can be found elsewhere [38]. Impedance measurements were calibrated using baseline solution measurement to compensate the frequency response of cable and electronics.…”

Section: Impedance Systemmentioning

confidence: 99%

“…On top of that, the fact that the two-electrode configuration includes the impedance and polarization of the counter and the working electrodes can jeopardize the data analysis if the contribution of each electrode is not the same. Furthermore, when the electrode surface is fully covered by bacteria or EPS, the inherent sensibility of this technique is significantly reduced as compared to the four-electrode configuration technique [38]. With the four-electrode method [55], the effect of the electrodeelectrolyte is excluded from the impedance measurement offering an increased sensitivity to monitor cell concentration and the possibility to extract morphological and structural information from the impedance (refer to Section 3.5).…”

Section: Impedance Monitoring Of Bacterial Biofilm In Growth Mediamentioning

confidence: 99%

“…To prevent the interpretation of the four-electrode impedance measurements from being influenced by ion concentration changes due to the biofilm metabolic activity [20] or temperature variation effects, we propose the use of a ratiometric biofilm estimator E following the same reasoning as in [38,66]. The estimator allows for obtaining a quantitative measurement of biofilm considering cells above the electrode surface.…”

Section: Biofilm Estimator Ementioning

confidence: 99%

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Simultaneous monitoring of Staphylococcus aureus growth in a multi-parametric microfluidic platform using microscopy and impedance spectroscopy

Estrada-Leypon

Moya

Guimerà‐Brunet

et al. 2015

Bioelectrochemistry

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“…In order to prevent direct contact Furthermore, electrode topology was specially conceived to perform bioimpedance measurements with a four-electrode configuration. Four-electrode measuring configurations are suitable for reducing the effects of the electrode polarization caused by the electrode-electrolyte interface [35], consequently improving the ability of the system to detect changes related to the cells [36,37]. In the present setup, we used a modified arrangement of a four-electrode measuring configuration by duplicating potential measurement terminals HP and LP (see Schematic 1c).…”

Section: Microelectrodesmentioning

confidence: 99%

Interpulse multifrequency electrical impedance measurements during electroporation of adherent differentiated myotubes

García-Sánchez

Azan

Leray

et al. 2015

Bioelectrochemistry

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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 longterm 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.

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Electrical impedance spectroscopy measurements using a four-electrode configuration improve on-line monitoring of cell concentration in adherent animal cell cultures

Cited by 42 publications

References 23 publications

Metabolic consequences of inflammatory disruption of the blood-brain barrier in an organ-on-chip model of the human neurovascular unit

Metabolic consequences of inflammatory disruption of the blood-brain barrier in an organ-on-chip model of the human neurovascular unit

Simultaneous monitoring of Staphylococcus aureus growth in a multi-parametric microfluidic platform using microscopy and impedance spectroscopy

Interpulse multifrequency electrical impedance measurements during electroporation of adherent differentiated myotubes

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