The field of bioelectronics is developing exponentially. There is now a drive to interface electronics with biology for the development of new technologies to improve our understanding of electrical forces in biology. This builds on our recently published work in which we show wireless electrochemistry could be used to grow bioelectronic functional circuitry in 2D cell layers. To date our ability to merge electronics with in situ with biology is 3D limited. In this study, we aimed to further develop the wireless electrochemical approach for the self-assembly of microwires in situ with custom-designed and fabricated 3D cancer spheroids. Unlike traditional electrochemical methods that rely on direct electrical connections to induce currents, our technique utilises bipolar electrodes that operate independently of physical wired connections. These electrodes enable redox reactions through the application of an external electric field. Specifically, feeder electrodes connected to a power supply generate an electric field, while the bipolar electrodes, not physically connected to the feeder electrodes, facilitate the reduction of silver ions from the solution. This process occurs upon applying a voltage across the feeder electrodes, resulting in the formation of self-assembled microwires between the cancer spheroids.Thereby, creating interlinked bioelectronic circuitry with cancer spheroids. We demonstrate that a direct current was needed to stimulate the growth of conductive microwires in the presence of cell spheroids. Microwire growth was successful when using 50 V (0.5 kV/cm) of DC applied to a single spheroid of approximately 800 µm in diameter but could not be achieved with alternating currents. This represents the first proof of the concept of using wireless electrochemistry to grow conductive structures with 3D mammalian cell spheroids.

A unique method for determining chlorophyll content in microalgae is devised employing a gold interdigitated electrode (G-IDE) with a 10-µm gap, augmented by a nano-molecularly imprinted polymer (nano-MIP) and a titanium dioxide/multiwalled carbon nanotube (TiO2/MWCNT) nanocomposite. The nano-MIP, produced using chlorophyll template voids, successfully trapped chlorophyll, while the TiO2/MWCNT nanocomposite, synthesized by the sol-gel technique, exhibited a consistent distribution and anatase crystalline structure. The rebinding of procured chlorophyll powder, which was used as a template for nano-MIP synthesis, was identified with a high determination coefficient (R2 = 0.9857). By combining the TiO2/MWCNT nanocomposite with nano-MIP, the G-IDE sensing method achieved a slightly better R2 value of 0.9892 for detecting chlorophyll in microalgae. The presented G-IDE sensor showed a significant threefold enhancement in chlorophyll detection compared with commercially available chlorophyll powder. It had a detection limit of 0.917 mL (v/v) and a linear range that spanned from 10-6 to 1 mL. The effectiveness of the sensor in detecting chlorophyll in microalgae was confirmed through validation of its repeatability and reusability.

Many bacteria perform extracellular electron transfer (EET), whereby electrons are transferred from the cell to an extracellular terminal electron acceptor. This electron acceptor can be an electrode and electrons can be delivered indirectly via a redox-active mediator molecule. Here, we present a protocol to study mediated EET in Lactiplantibacillus plantarum, a probiotic lactic acid bacterium widely used in the food industry, using a bioelectrochemical system. We detail how to assemble a three-electrode, two-chambered bioelectrochemical system and provide guidance on characterizing EET in the presence of a soluble mediator using chronoamperometry and cyclic voltammetry techniques. We use representative data from 1,4-dihydroxy-2-naphthoic acid (DHNA)-mediated EET experiments with L. plantarum to demonstrate data analysis and interpretation. The techniques described in this protocol can open new opportunities for electro-fermentation and bioelectrocatalysis. Recent applications of this electrochemical technique with L. plantarum demonstrated an acceleration of metabolic flux towards producing fermentation end-products, which are critical flavor components in food fermentation. As such, this system has the potential to be further developed to alter flavors in food production or produce valuable chemicals.

The present study elucidates the effectiveness of a molecularly imprinted polyacrylonitrile-imbued graphite-base electrode (MAN@G) for the selective detection of folic acid (FA) in food samples. The prime objective of the recognition and quantification of vitamin compounds like FA is the overall quality assessment of vegetables and fruits. The cost-effective, reproducible, and durable MAN@G electrode has been fabricated using acrylonitrile (AN) as the monomer and FA as the template over graphite-base. The characterization of the synthesized MAN@G electrode material has been accomplished by utilizing UV-visible (UV-vis) spectroscopy and scanning electron microscopy (SEM). A tri-electrode system based on differential pulse voltammetry (DPV) and cyclic voltammetry (CV) techniques was employed to explore the analytical performance of the synthesized electrode. Rigorous analyses divulged that a widespread linearity window could be exhibited by the electrode under an optimized experimental environment, ranging from 20 μM to 400 μM concentrations with an acceptable lower limit of detection (LOD) and limit of quantification (LOQ) of 18 nM, and 60 nM respectively. Additionally, this electrode exhibits high reproducibility, good stability, and high repeatability, with RSD values of 1.72 %, 1.32 %, and 1.19 %, respectively. The detection efficacy of the proposed electrode has been further examined in food extracts, namely orange, spinach, papaya, soybean, and cooked rice, which endorsed high accuracy compared to the high-performance liquid chromatography (HPLC) method. Moreover, the statistical results obtained from the t-test analysis were also satisfactory for the FA concentrations present in those five samples.

A wearable potentiometric device is reported based on an innovative butterfly-like paper-based microfluidic system, allowing for continuous monitoring of pH and Na+ levels in sweat during physical activity. Specifically, the use of the butterfly-like configuration avoids evaporation phenomena and memory effects, enabling precise and timely biomarker determination in sweat. Two ad hoc modified screen-printed electrodes were embedded in the butterfly-like paper-based microfluidics, and the sensing device was further integrated with a portable and miniaturized potentiostat, leveraging Bluetooth technology for efficient data transmission. First, the paper-based microfluidic configuration was tested for optimal fluidic management to obtain optimized performance of the device. Subsequently, the two electrodes were individually tested to detect the two biomarkers, namely pH and Na+. The results demonstrated highly promising near-Nernstian (0.056 ± 0.002 V/dec) and super-Nernstian (- 0.080 ± 0.003 V/pH) responses, for Na+ and pH detection, respectively. Additionally, several important parameters such as storage stability, interferents, and memory effect by hysteresis study were also investigated. Finally, the butterfly-like paper-based microfluidic wearable device was tested for Na+ and pH monitoring during the physical activity of three volunteers engaged in different exercises, obtaining a good correlation between Na+ increase and dehydration phenomena. Furthermore, one volunteer was tested through a cardiopulmonary test, demonstrating a correlation between sodium Na+ increase and the energetic effort by the volunteer. Our wearable device highlights the high potential to enable early evaluation of dehydration and open up new opportunities in sports activity monitoring.

Detecting dopamine (DA) in biological samples is vital to understand its crucial role in numerous physiological processes, such as motion, cognition, and reward stimulus. In this work, p-type graphene on sapphire, synthesized via chemical vapor deposition, serves as substrate for the preparation of p-type Cu2-xS films through solid-phase sulfurization. The optimized Cu2-xS/graphene heterostructure, prepared at 250 °C using a 15-nm copper film sulfurized for 2 h, exhibits superior electron transfer performance, ideal for electrochemical sensing. It is confirmed that the spontaneous charge transfer from graphene to Cu2-xS, higher Cu(II)/Cu(I) ratio (~ 0.8), and the presence of well-defined nanocrystalline structures with an average size of ~ 35 nm in Cu2-xS significantly contribute to the improved electron transfer of the heterostructure. The electrochemical sensor based on Cu2-xS/graphene heterostructure demonstrates remarkable sensitivity towards DA, with a detection limit as low as 100 fM and a dynamic range greater than 109 from 100 fM to 100 μM. Additionally, it exhibits excellent selectivity even in the presence of uric acid and ascorbic acid 100 times higher, alongside notable storage and measurement stability and repeatability. Impressively, the sensor also proves capable of detecting DA concentrations as low as 100 pM in rat serum, showcasing its potential for clinically relevant analytes and promising applications in sensitive, selective, reliable, and efficient point-of-care diagnostics.

Hydroquinone-based organic molecules are often used as unavoidable preservatives in the food industry. Among these additives, tertiary butylated hydroquinone (TBHQ) is widely employed as a preservative in various processed foods. However, the potential health risks associated with the excessive presence of TBHQ in food products have raised significant concerns. To address this pressing issuea novel binder-free composite composed of a manganese metal-organic framework and functionalized carbon nanofibers (Mn-MOF/f-CNF) has been developed as an electrode modifier for the ultrasensitive detection of TBHQ in food samples. The Mn-MOF/f-CNF composite was achieved using the ultrasonication method, revealing a lamellar sheet-like structure of the Mn-MOF and the curly thread-like fibrous structure of f-CNF. The developed Mn-MOF/f-CNF/SPE sensor system resulted in well-defined redox signals for TBHQ detection in a neutral pH solution. Compared to the unmodified SPE system, the modified system showed approximately a 300 mV reduction in overpotential and a twofold increase in peak current signal for TBHQ detection. The Mn-MOF/f-CNF/SPE sensor system showed a linear concentration window of 0.01 to 800 μM with a sensitivity of 6.28 µA µM-1 cm-2 and the obtained detection limit was 1.36 nM. Additionally, the proposed sensor displayed excellent reproducibility and repeatable results with an RSD of less than 5%. The real-time applicability of the Mn-MOF/f-CNF/SPE sensor system was demonstrated using real samples such as potato chips and instant noodles, showing excellent results with a recovery range of 95.1-98.5%.

Electrochemical approaches, along with miniaturization of electrodes, are increasingly being employed to detect and quantify nucleic acid biomarkers. Miniaturization of the electrodes is achieved through the use of screen-printed electrodes (SPEs), which consist of one to a few dozen sets of electrodes, or by utilizing printed circuit boards. Electrode materials used in SPEs include glassy carbon (Chiang H-C, Wang Y, Zhang Q, Levon K, Biosensors (Basel) 9:2-11, 2019), platinum, carbon, and graphene (Cheng FF, He TT, Miao HT, Shi JJ, Jiang LP, Zhu JJ, ACS Appl Mater Interfaces 7:2979-2985, 2015). There are numerous modifications to the electrode surfaces as well (Cheng FF, He TT, Miao HT, Shi JJ, Jiang LP, Zhu JJ, ACS Appl Mater Interfaces 7:2979-2985, 2015). These approaches offer distinct advantages, primarily due to their demonstrated superior limit of detection without amplification. Using the SPEs and potentiostats, we can detect cells, proteins, DNA, and RNA concentrations in the nanomolar (nM) to attomolar (aM) range. The focus of this chapter is to describe the basic approach adopted for the use of SPEs for nucleic acid measurement.

Cubic hollow-structured NiCo-LDH was synthesized using a solvothermal method. Subsequently, clay-like Ti3C2Tx MXenes were electrostatically self-assembled with NiCo layered double hydroxides (NiCo-LDH) to form composites featuring three-dimensional porous heterostructures. The composites were characterized using SEM, TEM, XRD, XPS, and FT-IR spectroscopy. Ti3C2Tx MXenes exhibit excellent electrical conductivity and hydrophilicity, providing abundant binding sites for NiCo-LDH, thereby promoting an increase in ion diffusion channels. The formation of three-dimensional porous heterostructural composites enhances charge transport, significantly improving sensor sensitivity and response speed. Consequently, the sensor demonstrates excellent electrochemical detection capability for quercetin (Qu), with a detection range of 0.1-20 µM and a detection limit of 23 nM. Additionally, it has been applied to the detection of Qu in natural plants such as onion, golden cypress, and chrysanthemum. The recovery ranged from 97.6 to 102.28%.

Metal ion-DNA interactions play a crucial role in modulating the structure and function of genetic material in the natural environment. In this study, we report on the favorable electrochemical activity of holmium(III) (Ho3+) on a glassy carbon electrode (GCE) and its interaction with double-stranded DNA. The interaction between DNA and Ho3+ was investigated for the first time using cyclic voltammetry and differential pulse voltammetry. The electrochemical behavior of Ho3+ ions on a GCE exhibited a reversible electron transfer process, indicative of its redox activity. A linear correlation between the peak current and the square root of the scan rate was observed, suggesting a diffusion-controlled kinetic regime for the electrochemical process. Additionally, fluorescence and absorption spectroscopy were employed to confirm the binding of Ho3+ to DNA. Our findings demonstrate that, at pH 7.2, specific DNA bases and phosphate groups can interact with Ho3+ ions. Moreover, electrochemical measurements suggest that Ho3+ ions bind to DNA via a groove binding mode, with a calculated binding ratio of 1:1 between Ho3+ and DNA. Notably, under optimal conditions, an increase in the amount of DNA leads to a significant reduction in the current intensity of Ho3+ ions.