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  • Bioelectrical signal detection using conducting polymer electrodes and the displacement current method
    Publication . Inácio, Pedro; Mestre, Ana L G; Medeiros, C.R.; Asgarifar, Sanaz; ELAMINE, Youssef; Canudo, Joana; Santos, João; Bragança, José; Morgado, Jorge; Biscarini, Fabio; Gomes, Henrique L.
    Conducting polymer electrodes based on poly (3, 4 ethylenedioxythiophene): polystyrene sulfonate were used to record electrophysiological signals from autonomous cardiac contractile cells present in embryoid bodies. Signal detection was carried out by measuring the displacement current across the polymer/electrolyte double-layer capacitance, and compared with voltage detection. While for relatively low capacitance electrodes, the voltage amplification provides higher signal quality, and for high capacitive electrodes, the displacement current method exhibits a higher signal-to-noise ratio. It is proposed that the displacement current method combined with high capacitive polymer-based electrodes is adequate to measure clusters of cells and whole organs. Our approach has a great potential in fundamental studies of drug discovery and safety pharmacology.
  • Ultrasensitive gold micro-structured electrodes enabling the detection of extra-cellular long-lasting potentials in astrocytes populations
    Publication . Mestre, Ana L. G.; Cerquido, Monica; INÁCIO, PEDRO; Asgarifar, Sanaz; Lourenco, Ana S.; Lurdes S. Cristiano, M.; Aguiar, Paulo; Medeiros, Maria C. R.; Araújo, Inês; Ventura, Joao; Gomes, Henrique L.
    Ultra-sensitive electrodes for extracellular recordings were fabricated and electrically characterized. A signal detection limit defined by a noise level of 0.3-0.4 mu V for a bandwidth of 12.5 Hz was achieved. To obtain this high sensitivity, large area (4 mm(2)) electrodes were used. The electrode surface is also micro-structured with an array of gold mushroom-like shapes to further enhance the active area. In comparison with a flat gold surface, the micro-structured surface increases the capacitance of the electrode/electrolyte interface by 54%. The electrode low impedance and low noise enable the detection of weak and low frequency quasi-periodic signals produced by astrocytes populations that thus far had remained inaccessible using conventional extracellular electrodes. Signals with 5 mu V in amplitude and lasting for 5-10 s were measured, with a peak-to-peak signal-to-noise ratio of 16. The electrodes and the methodology developed here can be used as an ultrasensitive electrophysiological tool to reveal the synchronization dynamics of ultra-slow ionic signalling between non-electrogenic cells.
  • Performance assessment of polymer based electrodes for in vitro electrophysiological sensing: the role of the electrode impedance
    Publication . Medeiros, Maria C. R.; Mestre, Ana L. G.; INÁCIO, PEDRO; Santos, João M. L.; Araújo, Inês; Bragança, José; Biscarini, Fabio; Gomes, Henrique L.
    Conducting polymer electrodes based on poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) are used to record extracellular signals from autonomous cardiac contractile cells and glioma cell cultures. The performance of these conducting polymer electrodes is compared with Au electrodes. A small-signal impedance analysis shows that in the presence of an electrolyte, both Au and polymer electrodes establish high capacitive double-layers. However, the polymer/electrolyte interfacial resistance is 3 orders of magnitude lower than the resistance of the metal/electrolyte interface. The polymer low interfacial resistance minimizes the intrinsic thermal noise and increases the system sensitivity. However, when measurements are carried out in current mode a low interfacial resistance partially acts as a short circuit of the interfacial capacitance, this affects the signal shape.
  • Extracellular electrophysiological measurements of cooperative signals in astrocytes populations
    Publication . Mestre, Ana L. G.; INÁCIO, PEDRO; ELAMINE, Youssef; Asgarifar, Sanaz; Lourenco, Ana S.; Cristiano, Maria L. S.; Aguiar, Paulo; Medeiros, Maria C. R.; Araújo, Inês; Ventura, Joao; Gomes, Henrique L.
    Astrocytes are neuroglial cells that exhibit functional electrical properties sensitive to neuronal activity and capable of modulating neurotransmission. Thus, electrophysiological recordings of astroglial activity are very attractive to study the dynamics of glial signaling. This contribution reports on the use of ultra-sensitive planar electrodes combined with low noise and low frequency amplifiers that enable the detection of extracellular signals produced by primary cultures of astrocytes isolated from mouse cerebral cortex. Recorded activity is characterized by spontaneous bursts comprised of discrete signals with pronounced changes on the signal rate and amplitude. Weak and sporadic signals become synchronized and evolve with time to higher amplitude signals with a quasi-periodic behavior, revealing a cooperative signaling process. The methodology presented herewith enables the study of ionic fluctuations of population of cells, complementing the single cells observation by calcium imaging as well as by patch-clamp techniques.
  • An ultra-low-cost RCL-meter
    Publication . Inácio, Pedro; Guerra, Rui; Stallinga, Peter
    An ultra-low-cost RCL meter, aimed at IoT applications, was developed, and was used to measure electrical components based on standard techniques without the need of additional electronics beyond the AVR® micro-controller hardware itself and high-level routines. The models and pseudo-routines required to measure admittance parameters are described, and a benchmark between the ATmega328P and ATmega32U4 AVR® micro-controllers was performed to validate the resistance and capacitance measurements. Both ATmega328P and ATmega32U4 micro-controllers could measure isolated resistances from 0.5 Ω to 80 MΩ and capacitances from 100 fF to 4.7 mF. Inductance measurements are estimated at between 0.2 mH to 1.5 H. The accuracy and range of the measurements of series and parallel RC networks are demonstrated. The relative accuracy (ar) and relative precision (pr) of the measurements were quantified. For the resistance measurements, typically ar, pr < 10% in the interval 100 Ω–100 MΩ. For the capacitance, measured in one of the modes (fast mode), ar < 20% and pr < 5% in the range 100 fF–10 nF, while for the other mode (transient mode), typically ar < 20% in the range 10 nF–10 mF and pr < 5% for 100 pF–10 mF. ar falls below 5% in some sub-ranges. The combination of the two capacitance modes allows for measurements in the range 100 fF–10 mF (11 orders of magnitude) with ar < 20%. Possible applications include the sensing of impedimetric sensor arrays targeted for wearable and in-body bioelectronics, smart agriculture, and smart cities, while complying with small form factor and low cost.
  • Electrical impedance spectroscopy for potassium content analysis and botanical origin identification of honey
    Publication . Elamine, Youssef; INÁCIO, PEDRO; Miguel, Maria da Graça; Carlier, Jorge; Costa, Maria Clara; Estevinho, Leticia M.; Gomes, Henrique L.
    Minerals are reported to dominate the electrical properties of honey and indicate its botanical and geographical origins. In this study, Electrochemical Impedance Spectroscopy (EIS) was used to assess the relation between mineral elements, electrical properties and botanical origin using three honey varieties - Citrus sp., Eucalyptus sp., and Erica sp. These varieties are identified through pollen analysis and market labelling. Flame atomic absorption and emission spectroscopies were used to quantify the concentrations of eight elements (potassium, sodium, calcium, magnesium, manganese, zinc, copper, and iron). Among all the mineral elements, potassium showed a consistent correlation with impedance. The potassium estimation in honey and standard solutions (calibration curve) had similar sensitivities of 153.43 nF/mM and 132.68 nF/mM, respectively. Additionally, the analysis revealed that potassium dominates the mineral composition, with the other species present in minimal quantities. The EIS technique showed high sensitivity to potassium and other ionisable species, making it possible to classify the botanical origin of these three honey types. The EIS technique proved to be both time and cost effective, yielding a classification rate higher than that achieved by analysing mineral composition.