Opportunities in optical and electrical single-cell technologies to study microbial ecosystems.

New techniques are revolutionizing single-cell research, allowing us to study microbes at unprecedented scales and in unparalleled depth. This review highlights the state-of-the-art technologies in single-cell analysis in microbial ecology applications, with particular attention to both optical tools, i.e., specialized use of flow cytometry and Raman spectroscopy and emerging electrical techniques. The objectives of this review include showcasing the diversity of single-cell optical approaches for studying microbiological phenomena, highlighting successful applications in understanding microbial systems, discussing emerging techniques, and encouraging the combination of established and novel approaches to address research questions. The review aims to answer key questions such as how single-cell approaches have advanced our understanding of individual and interacting cells, how they have been used to study uncultured microbes, which new analysis tools will become widespread, and how they contribute to our knowledge of ecological interactions.

Full-electric microfluidic platform to capture, analyze and selectively release single cells.

Current single-cell technologies require large and expensive equipment, limiting their use to specialized labs. In this paper, we present for the first time a microfluidic device which demonstrates a combined method for full-electric cell capturing, analyzing, and selectively releasing with single-cell resolution. All functionalities are experimentally demonstrated on Saccharomyces cerevisiae. Our microfluidic platform consists of traps centered around a pair of individually accessible coplanar electrodes, positioned under a microfluidic channel. Using this device, we validate our novel Two-Voltage method for trapping single cells by positive dielectrophoresis (pDEP). Cells are attracted to the trap when a high voltage (VH) is applied. A low voltage (VL) holds the already trapped cell in place without attracting additional cells, allowing full control over the number of trapped cells. After trapping, the cells are analyzed by broadband electrochemical impedance spectroscopy. These measurements allow the detection of single cells and the extraction of cell parameters. Additionally, these measurements show a strong correlation between average phase change and cell size, enabling the use of our system for size measurements in biological applications. Finally, our device allows selectively releasing trapped cells by turning off the pDEP signal in their trap. The experimental results show the techniques potential as a full-electric single-cell analysis tool with potential for miniaturization and automation which opens new avenues towards small-scale, high throughput single-cell analysis and sorting lab-on-CMOS devices.

Biodegradable Molybdenum (Mo) and Tungsten (W) Devices: One Step Closer towards Fully-Transient Biomedical Implants.

Close monitoring of vital physiological parameters is often key in following the evolution of certain medical conditions (e.g., diabetes, infections, post-operative status or post-traumatic injury). The allocation of trained medical staff and specialized equipment is, therefore, necessary and often translates into a clinical and economic burden on modern healthcare systems. As a growing field, transient electronics may establish fully bioresorbable medical devices capable of remote real-time monitoring of therapeutically relevant parameters. These devices could alert remote medical personnel in case of any anomaly and fully disintegrate in the body without a trace. Unfortunately, the need for a multitude of biodegradable electronic components (power supplies, wires, circuitry) in addition to the electrochemical biosensing interface has halted the arrival of fully bioresorbable electronically active medical devices. In recent years molybdenum (Mo) and tungsten (W) have drawn increasing attention as promising candidates for the fabrication of both energy-powered active (e.g., transistors and integrated circuits) and passive (e.g., resistors and capacitors) biodegradable electronic components. In this review, we discuss the latest Mo and W-based dissolvable devices for potential biomedical applications and how these soluble metals could pave the way towards next-generation fully transient implantable electronic systems.

Electrochemical Sensing of Adenosin Triphosphate by Specific Binding to Dipicolylamine Group in Cyclodextrin Supramolecular Complex.

Electrochemical detection based on cyclodextrin supramolecular complexes is founded on the competitive binding between electroactive probes and target molecules. This limits their versatility to be used for sensing a broad range of metabolites. In this work, we demonstrate the significant role of zinc ions as well as of ß-cyclodextrins modified with dipicolylamine and of a phenylboronic acid-modified ferrocene probe to address a selective electrochemical detection of adenosin triphosphate (ATP). Our findings will definitively have an impact in oncological point-of-care systems, since a high level of extracellular ATP reveals the inflammatory response due to chemotherapeutic treatments.

Electrochemical Assay for Extremely Selective Recognition of Fructose Based on 4-Ferrocene-Phenylboronic Acid Probe and ß-Cyclodextrins Supramolecular Complex.

The aim of the present paper is to highlight a novel electrochemical assay for an extremely-selective detection of fructose thanks to the use of a supramolecular complex between ß-cyclodextrins (ß-CDs) and a chemically modified ferrocene with boronic acid named 4-Fc-PB/natural-ß-CDs. Another kind of ß-CDs, the 4-Fc-PB/3-phenylboronic-ß-CDs, is proposed for the detection of glucose. The novel electrochemical probe is fully characterized by 1 H nuclear magnetic resonance, mass spectroscopy, and elemental analysis, while the superior electrochemical performance is assessed in terms of sensitivity and detection limit. The novelty of the present work consists in the role of CDs that for the first time are employed in electrochemistry with a unique detection mechanism based on specific chemical interactions with the target molecule by the introduction of proper binding groups. A highly selective detection of fructose is obtained and it is believed that the proposed mechanism of detection represents a new way to electrochemically sense other molecules by varying the combination of specific groups of the supramolecular complex. The findings are of impactful importance since a quick, easy, cheap, and extremely selective detection of fructose is not yet available in the market, here achieved by using electrochemical methods which are a very growing field.

Tracing the composition of single e-cigarette aerosol droplets in situ by laser-trapping and Raman scattering.

The partitioning of components between droplets and the gas phase in e-cigarette aerosols has a significant impact on deposition within the respiratory tract. However, exclusive detection of droplet composition has, so far, been elusive. Consequently, the dynamics of partitioning between droplets and the gas phase remains unknown. Here, we combine optical trapping of single droplets with in situ Raman scattering for destruction-free monitoring of e-cigarette droplet composition with a time resolution of seconds. We find that the initial droplet composition is very close to the composition of the e-liquid. Upon dilution with air, the droplet composition changes exponentially on a time scale of seconds, mainly because of evaporation of propylene glycol. The nicotine content in the droplet is controlled by the pH. Nicotine evaporates from the droplets under basic conditions, but remains in the liquid under acidic conditions. These results are crucial for advancing e-liquid research and manufacturing.

Fast Procedures for the Electrodeposition of Platinum Nanostructures on Miniaturized Electrodes for Improved Ion Sensing.

Nanostructured materials have attracted considerable interest over the last few decades to enhance sensing capabilities thanks to their unique properties and large surface area. In particular, noble metal nanostructures offer several advantages including high stability, non-toxicity and excellent electrochemical behaviour. However, in recent years the great expansion of point-of-care (POC) and wearable systems and the attempt to perform measurements in tiny spaces have also risen the need of increasing sensors miniaturization. Fast constant potential electrodeposition techniques have been proven to be an efficient way to obtain conformal platinum and gold nanostructured layers on macro-electrodes. However, this technique is not effective on micro-electrodes. In this paper, we investigate an alternative one-step deposition technique of platinum nanoflowers on micro-electrodes by linear sweep voltammetry (LSV). The effective deposition of platinum nanoflowers with similar properties to the ones deposited on macro-electrodes is confirmed by morphological analysis and by the similar roughness factor (~200) and capacitance (~18 µ F/mm 2 ). The electrochemical behaviour of the nanostructured layer is then tested in an solid-contact (SC) L i + -selective micro-electrode and compared to the case of macro-electrodes. The sensor offers Nernstian calibration with same response time (~15 s) and a one-order of magnitude smaller limit of detection (LOD) ( 2.6 × 10 - 6 ) with respect to the macro-ion-selective sensors (ISE). Finally, sensor reversibility and stability in both wet and dry conditions is proven.

A novel electrochemical sensor for non-invasive monitoring of lithium levels in mood disorders.

Lithium is the main drug for the treatment of mood disorders. Due to its narrow therapeutic window, Therapeutic Drug Monitoring (TDM) is a norm during therapy in order to avoid adverse effects. Consequently, patients are obliged to frequent check-ups in hospitals to determine their serum concentration and optimize accordingly the drug dose. Wearable sensors have attracted a growing interest in the research community in recent years owing to their promising impact in personalized healthcare. In particular, sweat diagnosis has seen an enormous expansion and is currently entering the market thanks to the large availability and simple collection of this fluid. In this paper a novel approach for non-invasive decentralized monitoring of lithium drug concentration through sweat analysis is proposed for the first time. An all-solid-state Ion- Selective Electrode (ISE) with a nanostructured Solid-Contact (SC) is used to detect lithium ions in sweat. The sensor offers near-Nernstian behaviour (57:6±2:1 mV/decade) in the concentration range of interest. In addition, it shows fast response (15-30 s), good reversibility and small potential drift over time. A wide pH stability window (pH 4-12) is also proved.

Highly-stable Li+ ion-selective electrodes based on noble metal nanostructured layers as solid-contacts.

Nowadays the development of stable and highly efficient Solid-Contact Ion-Selective Electrodes (SC-ISEs) attracts much attention in the research community because of the great expansion of portable analytical devices. In this work, we present highly stable Li+ all-solid-state ISEs exploiting noble metals nanostructures as ion-to-electron transducers. The detection of lithium is essential for therapeutic drug monitoring of bipolar patients. In addition, greater environmental exposure to this ion is occurring due to the large diffusion of lithium-ion batteries. However, only a limited number of SC Li+ ISEs already exists in literature based on Conductive Polymers (CPs) and carbon nanotubes. The use of noble metals for ion-to-electron transduction offers considerable advantages over CPs and carbon materials, including fast and conformal one-step deposition by electrochemical means, non-toxicity and high stability. We investigate for the first time the use of gold nanocorals obtained by means of a one-step electrodeposition process to improve sensor performance and we compare it to all-solid-state ISEs based on electrodeposited platinum nanoflowers. In addition, the effect of substrate electrode material, membrane thickness and conditioning concentration on the potentiometric response is carefully analysed. Scanning Electron Microscopy (SEM) and Current Reversal Chronopotentiometry (CRC) techniques are used to characterize the morphology and the electrochemical behaviour of the different ISEs. The use of nanostructured gold and platinum contacts allows the increase of the SC capacitance by one or two orders of magnitude, respectively, with respect to the flat metal, while the SC resistance is significantly reduced. We show that the microfabricated sensors offer Nernstian behaviour (58.7±0.8 mV/decade) in the activity range from 10-5 to 0.1 M, with short response time (∼15 s) and small potential drift during CRC measurements (dEdt=3×10-5±2×10-5 V/s). The exceptional response stability is verified also when no potential is applied. The sensor shows high selectivity towards all clinically important ions, with values very similar to conventional ISEs. Furthermore, to our knowledge, the selectivity towards Ca+2 is the best ever reported for SC-ISEs. In conclusion, the present study opens up new interesting perspectives towards the development of simple and reproducible fabrication protocols to obtain high-quality and high-stability all-solid-state ISEs.

A Differential Electrochemical Readout ASIC With Heterogeneous Integration of Bio-Nano Sensors for Amperometric Sensing.

A monolithic biosensing platform is presented for miniaturized amperometric electrochemical sensing in CMOS. The system consists of a fully integrated current readout circuit for differential current measurement as well as on-die sensors developed by growing platinum nanostructures (Pt-nanoS) on top of electrodes implemented with the top metal layer. The circuit is based on the switch-capacitor technique and includes pseudodifferential integrators for concurrent sampling of the differential sensor currents. The circuit further includes a differential to single converter and a programmable gain amplifier prior to an ADC. The system is fabricated in [Formula: see text] technology and measures current within [Formula: see text] with minimum input-referred noise of [Formula: see text] and consumes [Formula: see text] from a [Formula: see text] supply. Differential sensing for nanostructured sensors is proposed to build highly sensitive and offset-free sensors for metabolite detection. This is successfully tested for bio-nano-sensors for the measurement of glucose in submilli molar concentrations with the proposed readout IC. The on-die electrodes are nanostructured and cyclic voltammetry run successfully through the readout IC to demonstrate detection of [Formula: see text].

A bimetallic nanocoral Au decorated with Pt nanoflowers (bio)sensor for H2O2 detection at low potential.

In this work, we have developed for the first time a method to make novel gold and platinum hybrid bimetallic nanostructures differing in shape and size. Au-Pt nanostructures were prepared by electrodeposition in two simple steps. The first step consists of the electrodeposition of nanocoral Au onto a gold substrate using hydrogen as a dynamic template in an ammonium chloride solution. After that, the Pt nanostructures were deposited onto the nanocoral Au organized in pores. Using Pt (II) and Pt (IV), we realized nanocoral Au decorated with Pt nanospheres and nanocoral Au decorated with Pt nanoflowers, respectively. The bimetallic nanostructures showed better capability to electrochemically oxidize hydrogen peroxide compared with nanocoral Au. Moreover, Au-Pt nanostructures were able to lower the potential of detection and a higher performance was obtained at a low applied potential. Then, glucose oxidase was immobilized onto the bimetallic Au-Pt nanostructure using cross-linking with glutaraldehyde. The biosensor was characterized by chronoamperometry at +0.15V vs. Ag pseudo-reference electrode (PRE) and showed good analytical performances with a linear range from 0.01 to 2.00mM and a sensitivity of 33.66µA/mMcm2. The good value of Kmapp (2.28mM) demonstrates that the hybrid nanostructure is a favorable environment for the enzyme. Moreover, the low working potential can minimize the interference from ascorbic acid and uric acid as well as reducing power consumption to effect sensing. The simple procedure to realize this nanostructure and to immobilize enzymes, as well as the analytical performances of the resulting devices, encourage the use of this technology for the development of biosensors for clinical analysis.

Google Glass-Directed Monitoring and Control of Microfluidic Biosensors and Actuators.

Google Glass is a recently designed wearable device capable of displaying information in a smartphone-like hands-free format by wireless communication. The Glass also provides convenient control over remote devices, primarily enabled by voice recognition commands. These unique features of the Google Glass make it useful for medical and biomedical applications where hands-free experiences are strongly preferred. Here, we report for the first time, an integral set of hardware, firmware, software, and Glassware that enabled wireless transmission of sensor data onto the Google Glass for on-demand data visualization and real-time analysis. Additionally, the platform allowed the user to control outputs entered through the Glass, therefore achieving bi-directional Glass-device interfacing. Using this versatile platform, we demonstrated its capability in monitoring physical and physiological parameters such as temperature, pH, and morphology of liver- and heart-on-chips. Furthermore, we showed the capability to remotely introduce pharmaceutical compounds into a microfluidic human primary liver bioreactor at desired time points while monitoring their effects through the Glass. We believe that such an innovative platform, along with its concept, has set up a premise in wearable monitoring and controlling technology for a wide variety of applications in biomedicine.

Bubble electrodeposition of gold porous nanocorals for the enzymatic and non-enzymatic detection of glucose.

Au nanocorals are grown on gold screen-printed electrodes (SPEs) by using a novel and simple one-step electrodeposition process. Scanning electron microscopy was used for the morphological characterization. The devices were assembled on a three-electrode SPE system, which is flexible and mass producible. The electroactive surface area, determined by cyclic voltammetry in sulphuric acid, was found to be 0.07±0.01cm(2) and 35.3±2.7cm(2) for bare Au and nanocoral Au, respectively. The nanocoral modified SPEs were used to develop an enzymatic glucose biosensor based on H2O2 detection. Au nanocoral electrodes showed a higher sensitivity of 48.3±0.9µA/(mMcm(2)) at +0.45V vs Ag|AgCl compared to a value of 24.6±1.3µA/(mMcm(2)) at +0.70V vs Ag|AgCl obtained with bare Au electrodes. However, the modified electrodes have indeed proven to be extremely powerful for the direct detection of glucose with a non-enzymatic approach. The results confirmed a clear peak observed by using nanocoral Au electrode even in the presence of chloride ions at physiological concentration. Amperometric study carried out at +0.15V vs Ag|AgCl in the presence of 0.12M NaCl showed a linear range for glucose between 0.1 and 13mM.

Fast synthesis of platinum nanopetals and nanospheres for highly-sensitive non-enzymatic detection of glucose and selective sensing of ions.

Novel methods to obtain Pt nanostructured electrodes have raised particular interest due to their high performance in electrochemistry. Several nanostructuration methods proposed in the literature use costly and bulky equipment or are time-consuming due to the numerous steps they involve. Here, Pt nanostructures were produced for the first time by one-step template-free electrodeposition on Pt bare electrodes. The change in size and shape of the nanostructures is proven to be dependent on the deposition parameters and on the ratio between sulphuric acid and chloride-complexes (i.e., hexachloroplatinate or tetrachloroplatinate). To further improve the electrochemical properties of electrodes, depositions of Pt nanostructures on previously synthesised Pt nanostructures are also performed. The electroactive surface areas exhibit a two order of magnitude improvement when Pt nanostructures with the smallest size are used. All the biosensors based on Pt nanostructures and immobilised glucose oxidase display higher sensitivity as compared to bare Pt electrodes. Pt nanostructures retained an excellent electrocatalytic activity towards the direct oxidation of glucose. Finally, the nanodeposits were proven to be an excellent solid contact for ion measurements, significantly improving the time-stability of the potential. The use of these new nanostructured coatings in electrochemical sensors opens new perspectives for multipanel monitoring of human metabolism.

Efficient voltammetric discrimination of free bilirubin from uric acid and ascorbic acid by a CVD nanographite-based microelectrode.

We report a novel electrochemical sensor based on nanographite grown on platinum microelectrodes for the determination of bilirubin in the presence of normal concentrations of albumin. The albumin is a protein with an intrinsic ability to bind the bilirubin therefore reducing the concentration of the free electroactive metabolite in human fluids. In addition, the proposed device permits the discrimination of free bilirubin from two interferents, uric acid and ascorbic acid, by the separation of their oxidation peaks in voltammetry. Preliminary measurements in human serum prove that the proposed nanostructured platform can be used to detect bilirubin.

High-performance multipanel biosensors based on a selective integration of nanographite petals.

We report the first selective growth of nanographite petals and various carbon nanomaterials onto a multipanel electrochemical platform. Different types of nanomaterials can be obtained by fine-tuning the growth parameters of the chemical vapor deposition (CVD) process. First, absolute novelty is the catalytic CVD selective growth of different carbon nanomaterials only on the working electrodes of the platform. A second novelty is the growth obtained at complementary metal-oxide-semiconductor compatible temperatures. These novel electrodes have been incorporated in sensors in which performance characteristics improve with the content of nanostructures. Unprecedented sensing parameters with respect to both direct and enzyme-mediated electrochemical biodetection have been obtained.

Direct growth of nanotubes and graphene nanoflowers on electrochemical platinum electrodes.

Multi-walled carbon nanotubes and graphene nanoflowers were grown by a catalytic chemical vapor deposition process on metal surfaces. Electrodeposition was used as a versatile technique to obtain three different iron catalyst coatings on platinum microelectrodes. The influence of growth parameters on carbon deposits was investigated. Characterization was carried out by scanning electron microscopy and Raman spectroscopy. A chemical treatment in sulphuric acid produced an increased voltammetric background current. In Raman spectra, the effect of the chemical treatment is seen as a more pronounced sp(3) hybridisation mode of C resulting from surface functionalization of the C nanomaterials. Overall, the hybrid electrodes we produced exhibit a promising performance for oxidase-based array biosensors. Therefore, our study opens the possibility of integrating the hybrid electrodes in biochip applications.

Fully integrated biochip platforms for advanced healthcare.

Recent advances in microelectronics and biosensors are enabling developments of innovative biochips for advanced healthcare by providing fully integrated platforms for continuous monitoring of a large set of human disease biomarkers. Continuous monitoring of several human metabolites can be addressed by using fully integrated and minimally invasive devices located in the sub-cutis, typically in the peritoneal region. This extends the techniques of continuous monitoring of glucose currently being pursued with diabetic patients. However, several issues have to be considered in order to succeed in developing fully integrated and minimally invasive implantable devices. These innovative devices require a high-degree of integration, minimal invasive surgery, long-term biocompatibility, security and privacy in data transmission, high reliability, high reproducibility, high specificity, low detection limit and high sensitivity. Recent advances in the field have already proposed possible solutions for several of these issues. The aim of the present paper is to present a broad spectrum of recent results and to propose future directions of development in order to obtain fully implantable systems for the continuous monitoring of the human metabolism in advanced healthcare applications.