Optimization of Electrolytes with Redox Reagents to Improve the Impedimetric Signal for Use with a Low-Cost Analyzer.

The most well-known criterion for POC devices is ASSURED, and affordability, i.e., using low-cost instrumentation, is the most challenging one. This manuscript provides a pathway for transitioning ESSENCE, an impedance-based biosensor platform, from using an expensive benchtop analyzer-KeySight 4294A (~$50k)-to using a significantly portable and cheaper USB oscilloscope-Analog Discovery 2 (~$200) -with similar sensitivity (around 100 times price difference). To achieve this, we carried out a fundamental study of the interplay between an electrolyte like potassium chloride (KCl), and an electrolyte buffer like phosphate buffered saline (PBS) in the presence and absence of a redox buffer like ferro/ferricyanide system and ([Ru(bpy)3]2+). Redox molecules in the electrolyte caused a significant change in the Nyquist curve of the impedance depending on the redox molecule type. The redox species and the background electrolyte have their own RC semicircles in the Nyquist curve, whose overlap depends on the redox concentration and electrolyte ionic strength. We found that by increasing the electrolyte ionic strength or the redox concentration, the RC semicircle moves to higher frequencies and vice versa. Importantly, the use of the buffer electrolyte, instead of KCl, led to a lower standard deviation and overall signal (lesser sensitivity). However, to achieve the best results from the biorecognition signal, we chose a buffered electrolyte like PBS with high ionic strength and lowered the redox probe concentrations to minimize the standard deviation and reduce any noise from migrating to the low-cost analyzer. Comparing the two analyzers shows similar results, with a lowered detection limit from the low-cost analyzer.

Recent State and Challenges in Spectroelectrochemistry with Its Applications in Microfluidics.

This review paper presents the recent developments in spectroelectrochemical (SEC) technologies. The coupling of spectroscopy and electrochemistry enables SEC to do a detailed and comprehensive study of the electron transfer kinetics and vibrational spectroscopic fingerprint of analytes during electrochemical reactions. Though SEC is a promising technique, the usage of SEC techniques is still limited. Therefore, enough publicity for SEC is required, considering the promising potential in the analysis fields. Unlike previously published review papers primarily focused on the relatively frequently used SEC techniques (ultraviolet-visible SEC and surface-enhanced Raman spectroscopy SEC), the two not-frequently used but promising techniques (nuclear magnetic resonance SEC and dark-field microscopy SEC) have also been studied in detail. This review paper not only focuses on the applications of each SEC method but also details their primary working mechanism. In short, this paper summarizes each SEC technique's working principles, current applications, challenges encountered, and future development directions. In addition, each SEC technique's applicative research directions are detailed and compared in this review work. Furthermore, integrating SEC techniques into microfluidics is becoming a trend in minimized analysis devices. Therefore, the usage of SEC techniques in microfluidics is discussed.

A highly sensitive, easy-and-rapidly-fabricable microfluidic electrochemical cell with an enhanced three-dimensional electric field.

An NP-µFEC is a reusable, novel microfluidic electrochemical cell with multiple non-planar interdigitated microelectrode arrays, minimal sample volume, and enhanced electric field penetration for highly sensitive electrochemical analysis. (i) The NP-µFEC features spatial 3-electrode architecture, and a small sample volume (∼4 µL). (ii) Here, [Fe(CN)6]3-/4- redox couple are used as an electrochemical reporter. The effects on the electrochemical properties of NP-µFEC due to the change in the reference electrode (RE) and counter electrode (CE)'s position with respect to the working electrode (WE) position are analyzed. For NP-µFEC, the position of the RE with respect to the WE does not affect the CV, DPV electrochemical profiles. However, the spacing between the CE and WE plays a significant role. (iii) The enhanced three-dimensional electric field penetration in NP-µFEC is validated by finite element analysis simulation using COMSOL Multiphysics. (iv) Without electrode surface modifications, NP-µFEC shows a detection limit (DL) of ∼2.54 × 10-6 M for aqueous [Fe(CN)6]3-/4- probe. (v) The DL for Cu2+, Fe3+, and Hg2+ are 30.5±9.5 µg L-1, 181±58.5 µg L-1, and 12.4±1.95 µg L-1, respectively, which meets the US Environmental Protection Agency (EPA)'s water contamination level for Cu, Fe, and is close to that for Hg (EPA limits are 1300 µg L-1, 300 µg L-1, and 2 µg L-1, respectively). (vi) Further, using a pressure-sensitive adhesive layer to form the channel and create the NP-µFEC configuration simplifies the manufacturing process, making it cost-effective and allowing for rapid adoption in any research lab. NP-µFEC is used to detect heavy metal ions in water. This demonstrates that cost-effective, easy-to-fabricate NP-µFEC can be a new sensitive electrochemical platform.

Microfluidic Gas Sensors: Detection Principle and Applications.

With the rapid growth of emerging point-of-use (POU)/point-of-care (POC) detection technologies, miniaturized sensors for the real-time detection of gases and airborne pathogens have become essential to fight pollution, emerging contaminants, and pandemics. However, the low-cost development of miniaturized gas sensors without compromising selectivity, sensitivity, and response time remains challenging. Microfluidics is a promising technology that has been exploited for decades to overcome such limitations, making it an excellent candidate for POU/POC. However, microfluidic-based gas sensors remain a nascent field. In this review, the evolution of microfluidic gas sensors from basic electronic techniques to more advanced optical techniques such as surface-enhanced Raman spectroscopy to detect analytes is documented in detail. This paper focuses on the various detection methodologies used in microfluidic-based devices for detecting gases and airborne pathogens. Non-continuous microfluidic devices such as bubble/droplet-based microfluidics technology that have been employed to detect gases and airborne pathogens are also discussed. The selectivity, sensitivity, advantages/disadvantages vis-a-vis response time, and fabrication costs for all the microfluidic sensors are tabulated. The microfluidic sensors are grouped based on the target moiety, such as air pollutants such as carbon monoxide and nitrogen oxides, and airborne pathogens such as E. coli and SARS-CoV-2. The possible application scenarios for the various microfluidic devices are critically examined.

ESSENCE - A rapid, shear-enhanced, flow-through, capacitive electrochemical platform for rapid detection of biomolecules.

The rapid, sensitive, and selective detection of target analytes using electrochemical sensors is challenging. ESSENCE, a new Electrochemical Sensor that uses a Shear-Enhanced, flow-through Nanoporous Capacitive Electrode, overcomes current electrochemical sensors' response limitations, selectivity, and sensitivity limitations. ESSENCE is a microfluidic channel packed with transducer material sandwiched by a top and bottom microelectrode. The room-temperature instrument less integration process allows the switch of the transducer materials to make up the porous electrode without modifying the electrode architecture or device protocol. ESSENCE can be used to detect both biomolecules and small molecules by simply changing the packed transducer material. Electron microscopy results confirm the high porosity. In conjunction with the non-planar interdigitated electrode, the packed transducer material results in a flow-through porous electrode. Electron microscopy results confirm the high porosity. The enhanced shear forces and increased convective fluxes disrupt the electric double layer's (EDL) diffusive process in ESSENCE. This disruption migrates the EDL to high MHz frequency allowing the capture signal to be measured at around 100 kHz, significantly improving device timing (rapid detection) with a low signal-to-noise ratio. The device's unique architecture allows us multiple configuration modes for measuring the impedance signal. This allows us to use highly conductive materials like carbon nanotubes. We show that by combining single-walled carbon nanotubes as transducer material with appropriate capture probes, NP-µIDE has high selectivity and sensitivity for DNA (fM sensitivity, selective against non-target DNA), breast cancer biomarker proteins (p53, pg/L sensitivity, selective against non-target HER2).

Metal-Organic Framework-Based Microfluidic Impedance Sensor Platform for Ultrasensitive Detection of Perfluorooctanesulfonate.

The growing global concerns to public health from human exposure to perfluorooctanesulfonate (PFOS) require rapid, sensitive, in situ detection where current, state-of-the-art techniques are yet to adequately meet sensitivity standards of the real world. This work presents, for the first time, a synergistic approach for the targeted affinity-based capture of PFOS using a porous sorbent probe that enhances detection sensitivity by embedding it on a microfluidic platform. This novel sorbent-containing platform functions as an electrochemical sensor to directly measure PFOS concentration through a proportional change in electrical current (increase in impedance). The extremely high surface area and pore volume of mesoporous metal-organic framework (MOF) Cr-MIL-101 is used as the probe for targeted PFOS capture based on the affinity of the chromium center toward both the fluorine tail groups as well as the sulfonate functionalities as demonstrated by spectroscopic (NMR and XPS) and microscopic (TEM) studies. Answering the need for an ultrasensitive PFOS detection technique, we are embedding the MOF capture probes inside a microfluidic channel, sandwiched between interdigitated microelectrodes (IDµE). The nanoporous geometry, along with interdigitated microelectrodes, increases the signal-to-noise ratio tremendously. Further, the ability of the capture probes to interact with the PFOS at the molecular level and effectively transduce that response electrochemically has allowed us achieve a significant increase in sensitivity. The PFOS detection limit of 0.5 ng/L is unprecedented for in situ analytical PFOS sensors and comparable to quantification limits achieved using state-of-the-art ex situ techniques.

Effect of electrode configuration on the sensitivity of nucleic acid detection in a non-planar, flow-through, porous interdigitated electrode.

Electrical impedance spectroscopy (EIS) sensors, though rapid and cost-effective, often suffer from poor sensitivity. EIS sensors modified with carbon-based transducers show a higher conductance, thereby increasing the sensitivity of the sensor toward biomolecules such as DNA. However, the EIS spectra are compromised by the parasitic capacitance of the electric double layer (EDL). Here, a new shear-enhanced, flow-through nonporous, nonplanar interdigitated microelectrode sensor has been fabricated that shifts the EDL capacitor to high frequencies. Enhanced convective transport in this sensor disrupts the diffusion dynamics of the EDL, shifting its EIS spectra to high frequency. Concomitantly, the DNA detection signal shifts to high frequency, making the sensor very sensitive and rapid with a high signal to noise ratio. The device consists of a microfluidic channel sandwiched between two sets of top and bottom interdigitated microelectrodes. One of the sets of microelectrodes is packed with carbon-based transducer material such as carboxylated single-walled carbon nanotube (SWCNT). Multiple parametric studies of three different electrode configurations of the sensor along with different carbon-based transducer materials are undertaken to understand the fundamental physics and electrochemistry. Sensors packed with SWCNT show femtomolar detection sensitivity from all the different electrode configurations for a short target-DNA. A 20-fold jump in the signal is noticed from the unique working electrode configuration in contrast to the other electrode configurations. This demonstrates the potential of the sensor to have a significant increase in detection sensitivity for DNA and other biomolecules.

Probing the Sorption of Perfluorooctanesulfonate Using Mesoporous Metal-Organic Frameworks from Aqueous Solutions.

One approach to reduce increasing concentrations of toxic per- and polyfluoroalkyl substances (PFAS) involves the capture of PFAS from aqueous media using porous materials. The use of highly porous, tunable metal organic framework (MOF) materials is appealing for targeted liquid phase sorption. In this work, we demonstrate the excellent capture of perfluorooctanesulfonate (PFOS) using both the chromium and iron analogs of the MIL-101 framework. Experimental characterization of PFOS uptake reveals unique differences in sorption properties between these two analogs, providing key implications for future PFOS sorbent design. Specifically, STEM-EDS and IR spectroscopy show definitive proof of sorption. Furthermore, XPS analysis shows evidence of a strong interaction between sulfur atoms of the polar headgroup of PFOS and the metal center of the framework in addition to the fluorinated nonpolar tail. Additionally, in situ 19F NMR reveals higher PFOS affinity for Cr-MIL-101 versus Fe-MIL-101 based on sorption kinetics. Surprisingly, at these relatively high PFOS concentrations, activated acetylene black carbon is severely outperformed by both MOFs.

Cell Sequence and Mitosis Affect Fibroblast Directional Decision-Making During Chemotaxis in Microfluidic Mazes.

INTRODUCTION: Directed fibroblast migration is central to highly proliferative processes in regenerative medicine and developmental biology. However, the mechanisms by which single fibroblasts affect each other's directional decisions, while chemotaxing in microscopic pores, are not well understood. METHODS: We explored effects of cell sequence and mitosis on fibroblast platelet-derived growth factor-BB (PDGF-BB)-induced migration in microfluidic mazes with two possible through paths: short and long. Additionally, image-based modeling of the chemoattractant's diffusion, consumption and decay, was used to explain the experimental observations. RESULTS: It both cases, the cells displayed behavior that is contradictory to expectation based on the global chemoattractant gradient pre-established in the maze. In case of the sequence, the cells tend to alternate when faced with a bifurcation: if a leading cell takes the shorter (steeper gradient) path, the cell following it chooses the longer (weaker gradient) path, and vice versa. Image-based modeling of the process showed that the local PDGF-BB consumption by the individual fibroblasts may be responsible for this phenomenon. Additionally, it was found that when a mother cell divides, its two daughters go in opposite directions (even if it means migrating against the chemoattractant gradient and overcoming on-going cell traffic). CONCLUSIONS: It is apparent that micro-confined fibroblasts modify each other's directional decisions in a manner that is counter-intuitive to what is expected from classical chemotaxis theory. Consequently, accounting for these effects could lead to a better understanding of tissue generation in vivo, and result in more advanced engineered tissue products in vitro.

Plasmonic nano-protrusions: hierarchical nanostructures for single-molecule Raman spectroscopy.

Classical methods for enhancing the electromagnetic field from substrates for spectroscopic applications, such as surface-enhanced Raman spectroscopy (SERS), have involved the generation of hotspots through directed self-assembly of nanoparticles or by patterning nanoscale features using expensive nanolithography techniques. A novel large-area, cost-effective soft lithographic technique involving glancing angle deposition (GLAD) of silver on polymer gratings is reported here. This method produces hierarchical nanostructures with high enhancement factors capable of analyzing single-molecule SERS. The uniform ordered and patterned nanostructures provide extraordinary field enhancements that serve as excitatory hotspots and are herein interrogated by SERS. The high spatial homogeneity of the Raman signal and signal enhancement over a large area from a self-assembled monolayer (SAM) of 2-naphthalenethiol demonstrated the uniformity of the hotspots. The enhancement was shown to have a critical dependence on the underlying nanostructure via the surface energy landscape and GLAD angles for a fixed deposition thickness, as evidenced by atomic force microscopy and scanning electron microscopy surface analysis of the substrate. The nanostructured surface leads to an extremely concentrated electromagnetic field at sharp nanoscale peaks, here referred to as 'nano-protrusions', due to the coupling of surface plasmon resonance (SPR) with localized SPR. These nano-protrusions act as hotspots which provide Raman enhancement factors as high as 108 over a comparable SAM on silver. Comparison of our substrate with the commercial substrate Klarite™ shows higher signal enhancement and minimal signal variation with hotspot spatial distribution. By using the proper plasmon resonance angle corresponding to the laser source wavelength, further enhancement in signal intensity can be achieved. Single-molecule Raman spectra for rhodamine 6G are obtained from the best SERS substrate (a GLAD angle of 60°). The single-molecule spectrum is invariant over the substrate, due to the patterned ordered nanostructures (nano-protrusions).

A versatile self-assembly approach toward high performance nanoenergetic composite using functionalized graphene.

Exploiting the functionalization chemistry of graphene, long-range electrostatic and short-range covalent interactions were harnessed to produce multifunctional energetic materials through hierarchical self-assembly of nanoscale oxidizer and fuel into highly reactive macrostructures. Specifically, we report a methodology for directing the self-assembly of Al and Bi2O3 nanoparticles on functionalized graphene sheets (FGS) leading to the formation of nanocomposite structures in a colloidal suspension phase that ultimately condense into ultradense macrostructures. The mechanisms driving self-assembly were studied using a host of characterization techniques including zeta potential measurements, X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), particle size analysis, micro-Raman spectroscopy, and electron microscopy. A remarkable enhancement in energy release from 739 ± 18 to 1421 ± 12 J/g was experimentally measured for the FGS self-assembled nanocomposites.

A nanomembrane-based nucleic acid sensing platform for portable diagnostics.

In this perspective article, we introduce a potentially transformative DNA/RNA detection technology that promises to replace DNA microarray and real-time PCR for field applications. It represents a new microfluidic technology that fully exploits the small spatial dimensions of a biochip and some new phenomena unique to the micro- and nanoscales. More specifically, it satisfies all the requisites for portable on-field applications: fast, small, sensitive, selective, robust, label- and reagent-free, economical to produce, and possibly PCR-free. We discuss the mechanisms behind the technology and introduce some preliminary designs, test results, and prototypes.

Dynamic double layer effects on ac-induced dipoles of dielectric nanocolloids.

Normal and tangential surface ionic currents around low-permittivity nanocolloids with surface charges are shown to produce three different conductive mechanisms for ac-induced dipoles, all involving dynamic space charge accumulation at the double layerbulk interface with a conductivity jump. However, the distinct capacitor dimensions and diffusive contributions produce three disparate crossover frequencies at which the induced dipole reverses direction relative to the bulk field. A highly conducting collapsed diffuse layer, with bulk ion mobility, renders the particle conductive and produces an ionic strength independent crossover frequency for weak electrolytes. A precipitous drop in crossover frequency occurs at high ionic strengths when charging occurs only at the poles through field focusing around the insulated colloid. A peculiar maximum in crossover frequency exists between these two asymptotes for colloids smaller than a critical size when normal charging of the diffuse layer occurs over the entire surface. The crossover frequency data for latex nanocolloids of various sizes in different electrolytes of wide ranging ionic strengths are collapsed by explicit theoretical predictions without empirical parameters.

Designing a sensitive and quantifiable nanocolloid assay with dielectrophoretic crossover frequencies.

Dielectrophoretic nanocolloid assay is a promising technique for sensitive molecular detection and identification, as target molecule hybridization onto the probe-functionalized nanocolloids can change their surface conductance and consequently their dielectrophoretic crossover frequencies. Thus, instead of relying on surface charge density increase after hybridization, as in many capacitive and field effect transistor impedance sensing techniques, the current assay utilizes the much larger surface conductance (and dielectrophoresis crossover frequency) changes to effect sensitive detection. Herein, we present a Poisson-Boltzmann theory for surfaces with finite-size molecular probes that include the surface probe conformation, their contribution to surface charge with a proper delineation of the slip and Stern planes. The theory shows that the most sensitive nanocolloid molecular sensor corresponds to a minimum in the dielectrophoretic crossover frequency with respect to the bulk concentration of the molecular probes (oligonucleotides in our case) during nanocolloid functionalization. This minimum yields the lowest number of functionalized probes that are also fully stretched because of surface probe-probe interaction. Our theory provides the surface-bulk oligonucleotide concentration isotherm and a folding number for the surface oligonucleotide conformation from the crossover frequency, the zeta potential, and the hydrodynamic radius data.

A rapid field-use assay for mismatch number and location of hybridized DNAs.

Molecular dielectrophoresis (DEP) is employed to rapidly (

Identification and separation of DNA-hybridized nanocolloids by Taylor cone harmonics.

A rapid (minutes) electrospray bead-based DNA hybridization detection technique is developed by spraying a mixture of hybridized and unhybridized silica nanocolloids. With proper far-field control by external electrodes, the trajectory of the ejected nanobeads from the electrospray is governed by specific harmonics of the Laplace equation, which select discrete polar angles along well-separated field maxima near the conducting Taylor cone. Due to Rayleigh fission and evaporation, beads of different size acquire different total charge after ejection and suffer different normal electrophoretic displacement such that they are ejected along well-separated field maxima and are deposited in distinct rings on an intersecting plane. As the hybridized DNA is of the same dimension as that of the nanocolloid, the nanocolloids are hence easily differentiated from the unhybridized ones. This technique is highly specific as the high shear stress in the microjet shears away any non-specifically bound DNA from the nanocolloid surface.

Shear and AC Field Enhanced Carbon Nanotube Impedance Assay for Rapid, Sensitive, and Mismatch-Discriminating DNA Hybridization.

Other than concentrating the target molecules at the sensor location, we demonstrate two distinct new advantages of an open-flow impedance-sensing platform for DNA hybridization on carbon nanotube (CNT) surface in the presence of a high-frequency AC electric field. The shear-enhanced DNA and ion transport rate to the CNT surface decouples the parasitic double-layer AC impedance signal from the charge-transfer signal due to DNA hybridization. The flow field at high AC frequency also amplifies the charge-transfer rate across the hybridized CNT and provides shear-enhanced discrimination between DNA from targeted species and a closely related congeneric species with three nucleotide mismatches out of 26 bases in a targeted attachment region. This allows sensitive detection of hybridization events in less than 20 min with picomolar target DNA concentrations in a label-free CNT-based microfluidic detection platform.

Induced dipoles and dielectrophoresis of nanocolloids in electrolytes.

Electric induced dipoles of nanocolloids the size of the Debye length are shown to be one order stronger than predicted by the classical Maxwell-Wagner theory and its extensions. The difference is attributed to normal ion migration within the diffuse layer, and adsorption onto the Stern layer at the poles. The characteristic relaxation frequency (the crossover frequency for dielectrophoresis) is shown to be inversely proportional to the RC time of the diffuse layer capacitance and resistance, and has an anomalous -1 scaling with respect to the product of the Debye length and the particle size.