On-Chip Electrokinetic Micropumping for Nanoparticle Impact Electrochemistry.

Single-entity electrochemistry is a powerful technique to study the interactions of nanoparticles at the liquid-solid interface. In this work, we exploit Faradaic (background) processes in electrolytes of moderate ionic strength to evoke electrokinetic transport and study its influence on nanoparticle impacts. We implemented an electrode array comprising a macroscopic electrode that surrounds a set of 62 spatially distributed microelectrodes. This configuration allowed us to alter the global electrokinetic transport characteristics by adjusting the potential at the macroscopic electrode, while we concomitantly recorded silver nanoparticle impacts at the microscopic detection electrodes. By focusing on temporal changes of the impact rates, we were able to reveal alterations in the macroscopic particle transport. Our findings indicate a potential-dependent micropumping effect. The highest impact rates were obtained for strongly negative macroelectrode potentials and alkaline solutions, albeit also positive potentials lead to an increase in particle impacts. We explain this finding by reversal of the pumping direction. Variations in the electrolyte composition were shown to play a critical role as the macroelectrode processes can lead to depletion of ions, which influences both the particle oxidation and the reactions that drive the transport. Our study highlights that controlled on-chip micropumping is possible, yet its optimization is not straightforward. Nevertheless, the utilization of electro- and diffusiokinetic transport phenomena might be an appealing strategy to enhance the performance in future impact-based sensing applications.

Temperature profile characterization with fluorescence lifetime imaging microscopy in a thermophoretic chip.

This study introduces a thermophoretic lab-on-a-chip device to measure the Soret coefficient. We use resistive heating of a microwire on the chip to induce a temperature gradient, which is measured by fluorescence lifetime imaging microscopy (FLIM). To verify the functionality of the device, we used dyed polystyrene particles with a diameter of 25 nm. A confocal microscope is utilized to monitor the concentration profile of colloidal particles in the temperature field. Based on the measured temperature and concentration differences, we calculate the corresponding Soret coefficient. The same particles have been recently investigated with thermal diffusion forced Rayleigh scattering (TDFRS) and we find that the obtained Soret coefficients agree with literature results. This chip offers a simple way to study the thermophoretic behavior of biological systems in multicomponent buffer solutions quantitatively, which are difficult to study with optical methods solely relying on the refractive index contrast.

Inkjet-Printed and Electroplated 3D Electrodes for Recording Extracellular Signals in Cell Culture.

Recent investigations into cardiac or nervous tissues call for systems that are able to electrically record in 3D as opposed to 2D. Typically, challenging microfabrication steps are required to produce 3D microelectrode arrays capable of recording at the desired position within the tissue of interest. As an alternative, additive manufacturing is becoming a versatile platform for rapidly prototyping novel sensors with flexible geometric design. In this work, 3D MEAs for cell-culture applications were fabricated using a piezoelectric inkjet printer. The aspect ratio and height of the printed 3D electrodes were user-defined by adjusting the number of deposited droplets of silver nanoparticle ink along with a continuous printing method and an appropriate drop-to-drop delay. The Ag 3D MEAs were later electroplated with Au and Pt in order to reduce leakage of potentially cytotoxic silver ions into the cellular medium. The functionality of the array was confirmed using impedance spectroscopy, cyclic voltammetry, and recordings of extracellular potentials from cardiomyocyte-like HL-1 cells.

Nanoscale Electrochemical Sensor Arrays: Redox Cycling Amplification in Dual-Electrode Systems.

Micro- and nanofabriation technologies have a tremendous potential for the development of powerful sensor array platforms for electrochemical detection. The ability to integrate electrochemical sensor arrays with microfluidic devices nowadays provides possibilities for advanced lab-on-a-chip technology for the detection or quantification of multiple targets in a high-throughput approach. In particular, this is interesting for applications outside of analytical laboratories, such as point-of-care (POC) or on-site water screening where cost, measurement time, and the size of individual sensor devices are important factors to be considered. In addition, electrochemical sensor arrays can monitor biological processes in emerging cell-analysis platforms. Here, recent progress in the design of disease model systems and organ-on-a-chip technologies still needs to be matched by appropriate functionalities for application of external stimuli and read-out of cellular activity in long-term experiments. Preferably, data can be gathered not only at a singular location but at different spatial scales across a whole cell network, calling for new sensor array technologies. In this Account, we describe the evolution of chip-based nanoscale electrochemical sensor arrays, which have been developed and investigated in our group. Focusing on design and fabrication strategies that facilitate applications for the investigation of cellular networks, we emphasize the sensing of redox-active neurotransmitters on a chip. To this end, we address the impact of the device architecture on sensitivity, selectivity as well as on spatial and temporal resolution. Specifically, we highlight recent work on redox-cycling concepts using nanocavity sensor arrays, which provide an efficient amplification strategy for spatiotemporal detection of redox-active molecules. As redox-cycling electrochemistry critically depends on the ability to miniaturize and integrate closely spaced electrode systems, the fabrication of suitable nanoscale devices is of utmost importance for the development of this advanced sensor technology. Here, we address current challenges and limitations, which are associated with different redox cycling sensor array concepts and fabrication approaches. State-of-the-art micro- and nanofabrication technologies based on optical and electron-beam lithography allow precise control of the device layout and have led to a new generation of electrochemical sensor architectures for highly sensitive detection. Yet, these approaches are often expensive and limited to clean-room compatible materials. In consequence, they lack possibilities for upscaling to high-throughput fabrication at moderate costs. In this respect, self-assembly techniques can open new routes for electrochemical sensor design. This is true in particular for nanoporous redox cycling sensor arrays that have been developed in recent years and provide interesting alternatives to clean-room fabricated nanofluidic redox cycling devices. We conclude this Account with a discussion of emerging fabrication technologies based on printed electronics that we believe have the potential of transforming current redox cycling concepts from laboratory tools for fundamental studies and proof-of-principle analytical demonstrations into high-throughput devices for rapid screening applications.

The Influence of Supporting Ions on the Electrochemical Detection of Individual Silver Nanoparticles: Understanding the Shape and Frequency of Current Transients in Nano-impacts.

We report the influence of electrolyte composition and concentration on the stochastic amperometric detection of individual silver nanoparticles at microelectrode arrays and show that the sensor response at certain electrode potentials is dependent on both the conductivity of the electrolyte and the concentration of chloride ions. We further demonstrate that the chloride concentration in solution heavily influences the characteristic current spike shape of recorded nanoparticle impacts: While typically too short to be resolved in the measured current, the spike widths are significantly broadened at low chloride concentrations below 10 mm and range into the millisecond regime. The analysis of more than 25 000 spikes reveals that this effect can be explained by the diffusive mass transport of chloride ions to the nanoparticle, which limits the oxidation rate of individual silver nanoparticles to silver chloride at the chosen electrode potential.

Ion transport through polyelectrolyte multilayers.

Polyelectrolyte multilayer (PEM) films and capsules loaded with ion-sensitive fluorophores can be used as ion-sensors for many applications including measurements of intracellular ion concentration. Previous studies have shown the influence of the PEM films/shells on the specific response of encapsulated ion-sensitive fluorophores. PEM shells are considered as semipermeable barriers between the environment and the encapsulated fluorophores. Parameters such as the time response of the encapsulated sensor can be affected by the porosity and charge of the PEM shell. In this study, the time response of an encapsulated pH-sensitive fluorophore towards pH changes in the surrounding environment is investigated. Furthermore, the conductance of PEM films for potassium ions is determined.

Inkjet-printed 3D micro-ring-electrode arrays for amperometric nanoparticle detection.

Chip-based impact electrochemistry can provide means to measure nanoparticles in solution by sensing their stochastic collisions on appropriately-polarized microelectrodes. However, a planar microelectrode array design still restricts the particle detection to the chip surface and does not allow detection in 3D environments. In this work, we report a fast fabrication process for 3D microelectrode arrays by combining ink-jet printing with laser-patterning. To this end, we printed 3D pillars from polyacrylate ink as a scaffold. Then, the metal structures are manufactured via sputtering and laser-ablation. Finally, the chip is passivated with a parylene-C layer and the electrode tips are created via laser-ablation in a vertical alignment. As a proof of principle, we employ our 3D micro-ring-electrode arrays for single impact recordings from silver nanoparticles.

Influence of Auditory Cues on the Neuronal Response to Naturalistic Visual Stimuli in a Virtual Reality Setting.

Virtual reality environments offer great opportunities to study the performance of brain-computer interfaces (BCIs) in real-world contexts. As real-world stimuli are typically multimodal, their neuronal integration elicits complex response patterns. To investigate the effect of additional auditory cues on the processing of visual information, we used virtual reality to mimic safety-related events in an industrial environment while we concomitantly recorded electroencephalography (EEG) signals. We simulated a box traveling on a conveyor belt system where two types of stimuli - an exploding and a burning box - interrupt regular operation. The recordings from 16 subjects were divided into two subsets, a visual-only and an audio-visual experiment. In the visual-only experiment, the response patterns for both stimuli elicited a similar pattern - a visual evoked potential (VEP) followed by an event-related potential (ERP) over the occipital-parietal lobe. Moreover, we found the perceived severity of the event to be reflected in the signal amplitude. Interestingly, the additional auditory cues had a twofold effect on the previous findings: The P1 component was significantly suppressed in the case of the exploding box stimulus, whereas the N2c showed an enhancement for the burning box stimulus. This result highlights the impact of multisensory integration on the performance of realistic BCI applications. Indeed, we observed alterations in the offline classification accuracy for a detection task based on a mixed feature extraction (variance, power spectral density, and discrete wavelet transform) and a support vector machine classifier. In the case of the explosion, the accuracy slightly decreased by -1.64% p. in an audio-visual experiment compared to the visual-only. Contrarily, the classification accuracy for the burning box increased by 5.58% p. when additional auditory cues were present. Hence, we conclude, that especially in challenging detection tasks, it is favorable to consider the potential of multisensory integration when BCIs are supposed to operate under (multimodal) real-world conditions.

Single-Impact Electrochemistry in Paper-Based Microfluidics.

Microfluidic paper-based analytical devices (µPADs) have experienced an unprecedented story of success. In particular, as of today, most people have likely come into contact with one of their two most famous examples─the pregnancy or the SARS-CoV-2 antigen test. However, their sensing performance is constrained by the optical readout of nanoparticle agglomeration, which typically allows only qualitative measurements. In contrast, single-impact electrochemistry offers the possibility to quantify species concentrations beyond the pM range by resolving collisions of individual species on a microelectrode. Within this work, we investigate the integration of stochastic sensing into a µPAD design by combining a wax-patterned microchannel with a microelectrode array to detect silver nanoparticles (AgNPs) by their oxidative dissolution. In doing so, we demonstrate the possibility to resolve individual nanoparticle collisions in a reference-on-chip configuration. To simulate a lateral flow architecture, we flush previously dried AgNPs along a microchannel toward the electrode array, where we are able to record nanoparticle impacts. Consequently, single-impact electrochemistry poses a promising candidate to extend the limits of lateral flow-based sensors beyond current applications toward a fast and reliable detection of very dilute species on site.

Prototype Digital Lateral Flow Sensor Using Impact Electrochemistry in a Competitive Binding Assay.

This work demonstrates a lateral flow assay concept on the basis of stochastic-impact electrochemistry. To this end, we first elucidate requirements to employ silver nanoparticles as redox-active labels. Then, we present a prototype that utilizes nanoimpacts from biotinylated silver nanoparticles as readouts to detect free biotin in solution based on competitive binding. The detection is performed in a membrane-based microfluidic system, where free biotin and biotinylated particles compete for streptavidin immobilized on embedded latex beads. Excess nanoparticles are then registered downstream at an array of detection electrodes. In this way, we establish a proof of concept that serves as a blueprint for future "digital" lateral flow sensors.

Fully Printed µ-Needle Electrode Array from Conductive Polymer Ink for Bioelectronic Applications.

Microelectrode arrays (MEAs) are widely used platforms in bioelectronics to study electrogenic cells. In recent years, the processing of conductive polymers for the fabrication of three-dimensional electrode arrays has gained increasing interest for the development of novel sensor designs. Here, additive manufacturing techniques are promising tools for the production of MEAs with three-dimensional electrodes. In this work, a facile additive manufacturing process for the fabrication of MEAs that feature needle-like electrode tips, so-called µ-needles, is presented. To this end, an aerosol-jet compatible PEDOT:PSS and multiwalled carbon nanotube composite ink with a conductivity of 323 ± 75 S m-1 is developed and used in a combined inkjet and aerosol-jet printing process to produce the µ-needle electrode features. The µ-needles are fabricated with a diameter of 10 ± 2 µm and a height of 33 ± 4 µm. They penetrate an inkjet-printed dielectric layer to a height of 12 ± 3 µm. After successful printing, the electrochemical properties of the devices are assessed via cyclic voltammetry and impedance spectroscopy. The µ-needles show a capacitance of 242 ± 70 nF at a scan rate of 5 mV s-1 and an impedance of 128 ± 22 kΩ at 1 kHz frequency. The stability of the µ-needle MEAs in aqueous electrolyte is demonstrated and the devices are used to record extracellular signals from cardiomyocyte-like HL-1 cells. This proof-of-principle experiment shows the µ-needle MEAs' cell-culture compatibility and functional integrity to investigate electrophysiological signals from living cells.

On-Chip Stochastic Detection of Silver Nanoparticles without a Reference Electrode.

We report the electrochemical detection of 20 nm silver nanoparticles at a chip-based microelectrode array (MEA) without the need for a conventional reference electrode. This is possible due to the system's open-circuit potential allowing the oxidation of silver nanoparticles in the presence of phosphate-buffered saline (PBS). The hypothesis is confirmed by modulating the open-circuit potential via addition of ascorbic acid in solution, effectively inhibiting the detection of silver nanoparticle events. Employing the reference-free detection concept, we observe a linear relationship between the nanoparticle impact frequency at the microelectrodes and the nanoparticle concentration. This allows for viable quantification of silver nanoparticle concentrations in situ. The presented concept is ideal for the development of simple lab-on-a-chip or point-of-use systems enabling fast and low-cost screening of nanoparticles.

On-chip electromagnetic tweezers - 3-dimensional particle actuation using microwire crossbar arrays.

Emerging miniaturization technologies for biological and bioengineering applications require precise control over position and actuation of microparticles. While many of these applications call for high-throughput approaches, common tools for particle manipulation, such as magnetic or optical tweezers, suffer from low parallelizability. To address this issue, we introduce a chip-based platform that enables flexible three-dimensional control over individual magnetic microparticles. Our system relies on microwire crossbar arrays for simultaneous generation of magnetic and dielectric forces, which actuate the particles along highly localized traps. We demonstrate the precise spatiotemporal control of individual particles by tracing complex trajectories in three dimensions and investigate the forces that can be generated along different axes. Furthermore, we show that our approach for particle actuation can be parallelized by simultaneously controlling the position and movement of 16 particles in parallel.

Inducing microscopic thermal lesions for the dissection of functional cell networks on a chip.

We present a versatile chip-based method to inflict microscopic lesions on cellular networks or tissue models. Our approach relies on resistive heating of microstructured conductors to impose highly localized thermal stress on specific regions of a cell network. We show that networks can be precisely dissected into individual subnetworks using a microwire crossbar array. To this end, we pattern a network of actively beating cardiomyocyte-like cells into smaller subunits by inflicting thermal damage along selected wires of the array. We then investigate the activity and functional connectivity of the individual subnetworks using a Ca(2+) imaging-based signal propagation analysis. Our results demonstrate the efficient separation of functional activity between individual subnetworks on a microscopic level. We believe that the presented technique may become a powerful tool for investigating lesion and regeneration models in cellular networks.

Nanostructured cavity devices for extracellular stimulation of HL-1 cells.

Microelectrode arrays (MEAs) are state-of-the-art devices for extracellular recording and stimulation on biological tissue. Furthermore, they are a relevant tool for the development of biomedical applications like retina, cochlear and motor prostheses, cardiac pacemakers and drug screening. Hence, research on functional cell-sensor interfaces, as well as the development of new surface structures and modifications for improved electrode characteristics, is a vivid and well established field. However, combining single-cell resolution with sufficient signal coupling remains challenging due to poor cell-electrode sealing. Furthermore, electrodes with diameters below 20 µm often suffer from a high electrical impedance affecting the noise during voltage recordings. In this study, we report on a nanocavity sensor array for voltage-controlled stimulation and extracellular action potential recordings on cellular networks. Nanocavity devices combine the advantages of low-impedance electrodes with small cell-chip interfaces, preserving a high spatial resolution for recording and stimulation. A reservoir between opening aperture and electrode is provided, allowing the cell to access the structure for a tight cell-sensor sealing. We present the well-controlled fabrication process and the effect of cavity formation and electrode patterning on the sensor's impedance. Further, we demonstrate reliable voltage-controlled stimulation using nanostructured cavity devices by capturing the pacemaker of an HL-1 cell network.