Biosensors, Volume 6, Issue 4 (December 2016) – 13 articles

Portable detection and quantitation methods for Bacillus anthracis (anthrax) spores in pure culture or in environmental samples are lacking. Here, an amperometric immunoassay has been developed utilizing immunomagnetic separation to capture the spores and remove potential interferents from test samples followed by amperometric measurement on a field-portable instrument. Antibody-conjugated magnetic beads and antibody-conjugated glucose oxidase were used in a sandwich format for the capture and detection of target spores. Glucose oxidase activity of spore pellets was measured indirectly via amperometry by applying a bias voltage after incubation with glucose, horseradish peroxidase, and the electron mediator 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid). Target capture was mediated by polyclonal antisera, whereas monoclonal antibodies were used for signal generation. This strategy maximized sensitivity (500 target spores, 5000 cfu/mL), while also providing a good specificity for Bacillus anthracis spores. Minimal signal deviation occurs in the presence of environmental interferents including soil and modified pH conditions, demonstrating the strengths of immunomagnetic separation. The simultaneous incubation of capture and detection antibodies and rapid substrate development (5 min) result in short sample-to-signal times (less than an hour). With attributes comparable or exceeding that of ELISA and LFDs, amperometry is a low-cost, low-weight, and practical method for detecting anthrax spores in the field. Full article

To determine the originality of a typical Italian Parmigiano Reggiano cheese, it is crucial to define and characterize its quality, ripening period, and geographical origin. Different analytical techniques have been applied aimed at studying the organoleptic and characteristic volatile organic compounds (VOCs) profile of this cheese. However, most of the classical methods are time consuming and costly. The aim of this work was to illustrate a new simple, portable, fast, reliable, non-destructive, and economic sensor device S3 based on an array of six metal oxide semiconductor nanowire gas sensors to assess and discriminate the quality ranking of grated Parmigiano Reggiano cheese samples and to identify the VOC biomarkers using a headspace SPME-GC-MS. The device could clearly differentiate cheese samples varying in quality and ripening time when the results were analyzed by multivariate statistical analysis involving principal component analysis (PCA). Similarly, the volatile constituents of Parmigiano Reggiano identified were consistent with the compounds intimated in the literature. The obtained results show the applicability of an S3 device combined with SPME-GC-MS and sensory evaluation for a fast and high-sensitivity analysis of VOCs in Parmigiano Reggiano cheese and for the quality control of this class of cheese. Full article

The propagation of surface acoustic Love modes along ZnO/glass-based structures was modeled and analysed with the goal of designing a sensor able to detect changes in the environmental parameters, such as liquid viscosity changes and minute amounts of mass supported in the viscous liquid medium. Love mode propagation was modeled by numerically solving the system of coupled electro-mechanical field equations and Navier–Stokes equations. The phase and group velocities and the attenuation of the acoustic wave propagating along the 30° tilted c-axis ZnO/glass structure contacting a viscous non-conductive liquid were calculated for different ZnO guiding layer thicknesses, added mass thicknesses, and liquid viscosity and density. The three sensor responses, i.e., the wave phase and group velocity, and attenuation changes are calculated for different environmental parameters and related to the sensor velocity and attenuation sensitivities. The resulted sensitivities to liquid viscosity and added mass were optimized by adjusting the ZnO guiding layer thickness corresponding to a sensitivity peak. The present analysis is valuable for the manufacture and application of the ZnO-glass structure Love wave sensors for the detection of liquid properties, such as viscosity, density and mass anchored to the sensor surface. Full article

Gait analysis using wearable wireless sensors can be an economical, convenient and effective way to provide diagnostic and clinical information for various health-related issues. In this work, our custom designed low-cost wireless gait analysis sensor that contains a basic inertial measurement unit (IMU) was used to collect the gait data for four patients diagnosed with balance disorders and additionally three normal subjects, each performing the Dynamic Gait Index (DGI) tests while wearing the custom wireless gait analysis sensor (WGAS). The small WGAS includes a tri-axial accelerometer integrated circuit (IC), two gyroscopes ICs and a Texas Instruments (TI) MSP430 microcontroller and is worn by each subject at the T4 position during the DGI tests. The raw gait data are wirelessly transmitted from the WGAS to a near-by PC for real-time gait data collection and analysis. In order to perform successful classification of patients vs. normal subjects, we used several different classification algorithms, such as the back propagation artificial neural network (BP-ANN), support vector machine (SVM), k-nearest neighbors (KNN) and binary decision trees (BDT), based on features extracted from the raw gait data of the gyroscopes and accelerometers. When the range was used as the input feature, the overall classification accuracy obtained is 100% with BP-ANN, 98% with SVM, 96% with KNN and 94% using BDT. Similar high classification accuracy results were also achieved when the standard deviation or other values were used as input features to these classifiers. These results show that gait data collected from our very low-cost wearable wireless gait sensor can effectively differentiate patients with balance disorders from normal subjects in real time using various classifiers, the success of which may eventually lead to accurate and objective diagnosis of abnormal human gaits and their underlying etiologies in the future, as more patient data are being collected. Full article

Breast cancer is the most prevalent cause of cancer-associated death in women the world over, but if detected early it can be treated successfully. Therefore, it is important to diagnose this disease at an early stage and to understand the biochemical changes associated with cellular transformation and cancer progression. Deregulated lipid metabolism has been shown to contribute to cell transformation as well as cancer progression. In this study, we monitored the biomolecular changes associated with the transformation of a normal cell into an invasive cell associated with breast cancer using Raman microspectroscopy. We have utilized primary normal breast cells, and immortalized, transformed, non-invasive, and invasive breast cancer cells. The Raman spectra were acquired from all these cell lines under physiological conditions. The higher wavenumber (2800–3000 cm⁻¹) and lower wavenumber (700–1800 cm⁻¹) range of the Raman spectrum were analyzed and we observed increased lipid levels for invasive cells. The Raman spectral data were analyzed by principal component–linear discriminant analysis (PC-LDA), which resulted in the formation of distinct clusters for different cell types with a high degree of sensitivity. The subsequent testing of the PC-LDA analysis via the leave-one-out cross validation approach (LOOCV) yielded relatively high identification sensitivity. Additionally, the Raman spectroscopic results were confirmed through fluorescence staining tests with BODIPY and Nile Red biochemical assays. Furthermore, Raman maps from the above mentioned cells under fixed conditions were also acquired to visualize the distribution of biomolecules throughout the cell. The present study shows the suitability of Raman spectroscopy as a non-invasive, label-free, microspectroscopic technique, having the potential of probing changes in the biomolecular composition of living cells as well as fixed cells. Full article

An amperometric magnetoimmunosensor for the determination of human p53 protein is described in this work using a sandwich configuration involving the covalent immobilization of a specific capture antibody onto activated carboxylic-modified magnetic beads (HOOC-MBs) and incubation of the modified MBs with a mixture of the target protein and horseradish peroxidase-labeled antibody (HRP-anti-p53). The resulting modified MBs are captured by a magnet placed under the surface of a disposable carbon screen-printed electrode (SPCE) and the amperometric responses are measured at −0.20 V (vs. an Ag pseudo-reference electrode), upon addition of hydroquinone (HQ) as a redox mediator and H₂O₂ as the enzyme substrate. The magnetoimmunosensing platform was successfully applied for the detection of p53 protein in different cell lysates without any matrix effect after a simple sample dilution. The results correlated accurately with those provided by a commercial ELISA kit, thus confirming the immunosensor as an attractive alternative for rapid and simple determination of this protein using portable and affordable instrumentation. Full article

Biomedical signals contain features that represent physiological events, and each of these events has peaks. The analysis of biomedical signals for monitoring or diagnosing diseases requires the detection of these peaks, making event detection a crucial step in biomedical signal processing. Many researchers have difficulty detecting these peaks to investigate, interpret and analyze their corresponding events. To date, there is no generic framework that captures these events in a robust, efficient and consistent manner. A new method referred to for the first time as two event-related moving averages (“TERMA”) involves event-related moving averages and detects events in biomedical signals. The TERMA framework is flexible and universal and consists of six independent LEGO building bricks to achieve high accuracy detection of biomedical events. Results recommend that the window sizes for the two moving averages ( $W_1$ and $W_2$ ) have to follow the inequality $(8\times W_1)\ge W_2\ge (2\times W_1)$ . Moreover, TERMA is a simple yet efficient event detector that is suitable for wearable devices, point-of-care devices, fitness trackers and smart watches, compared to more complex machine learning solutions. Full article

To achieve sensitivity, comfort, and durability in vital sign monitoring, this study explores the use of radar technologies in wearable devices. The study first detected the respiratory rates and heart rates of a subject at a one-meter distance using a self-injection-locked (SIL) radar and a conventional continuous-wave (CW) radar to compare the sensitivity versus power consumption between the two radars. Then, a pulse rate monitor was constructed based on a bistatic SIL radar architecture. This monitor uses an active antenna that is composed of a SIL oscillator (SILO) and a patch antenna. When attached to a band worn on the subject’s wrist, the active antenna can monitor the pulse on the subject’s wrist by modulating the SILO with the associated Doppler signal. Subsequently, the SILO’s output signal is received and demodulated by a remote frequency discriminator to obtain the pulse rate information. Full article

Extracellular potassium concentration, [K⁺]_o, plays a fundamental role in the physiological functions of the brain. Studies investigating changes in [K⁺]_o have predominantly relied upon glass capillary electrodes with K⁺-sensitive solution gradients for their measurements. However, such electrodes are unsuitable for taking spatio-temporal measurements and are limited by the surface area of their tips. We illustrate seizures invoked chemically and in optogenetically modified mice using blue light exposure while impedimetrically measuring the response. A sharp decrease of 1–2 mM in [K⁺]_o before each spike has shown new physiological events not witnessed previously when measuring extracellular potassium concentrations during seizures in mice. We propose a novel approach that uses multichannel monolayer coated gold microelectrodes for in vivo spatio-temporal measurements of [K⁺]_o in a mouse brain as an improvement to the conventional glass capillary electrode. Full article

Optical biosensors based on photonic crystal surface waves (PC SWs) offer a possibility to study binding interactions with living cells, overcoming the limitation of rather small evanescent field penetration depth into a sample medium that is characteristic for typical optical biosensors. Besides this, simultaneous excitation of s- and p-polarized surface waves with different penetration depths is realized here, permitting unambiguous separation of surface and volume contributions to the measured signal. PC-based biosensors do not require a bulk signal correction, compared to widely used surface plasmon resonance-based devices. We developed a chitosan-based protocol of PC chip functionalization for bacterial attachment and performed experiments on antibody binding to living bacteria measured in real time by the PCSW-based biosensor. Data analysis reveals specific binding and gives the value of the dissociation constant for monoclonal antibodies (IgG2b) against bacterial lipopolysaccharides equal to K_D = 6.2 ± 3.4 nM. To our knowledge, this is a first demonstration of antibody-binding kinetics to living bacteria by a label-free optical biosensor. Full article

From the bench-mark work on microfluidics from the Whitesides’s group in 2007, paper technology has experienced significant growth, particularly regarding applications in biomedical research and clinical diagnostics. Besides the structural properties supporting microfluidics, other advantageous features of paper materials, including their versatility, disposability and low cost, show off the great potential for the development of advanced and eco-friendly analytical tools. Consequently, paper was quickly employed in the field of electrochemical sensors, being an ideal material for producing custom, tailored and miniaturized devices. Stencil-, inkjet-, or screen-printing are the preferential techniques for electrode manufacturing. Not surprisingly, we witnessed a rapid increase in the number of publications on paper based screen-printed sensors at the turn of the past decade. Among the sensing strategies, various biosensors, coupling electrochemical detectors with biomolecules, have been proposed. This work provides a critical review and a discussion on the future progress of paper technology in the context of miniaturized printed electrochemical biosensors. Full article

This review describes recent advances in the fabrication of electrochemical (bio)sensors based on screen-printing technology involving carbon materials and their application in biomedical, agri-food and environmental analyses. It will focus on the various strategies employed in the fabrication of screen-printed (bio)sensors, together with their performance characteristics; the application of these devices for the measurement of selected naturally occurring biomolecules, environmental pollutants and toxins will be discussed. Full article

Enteric and diarrheal diseases are a major cause of childhood illness and death in countries with developing economies. Each year, more than half of a million children under the age of five die from these diseases. We have developed a portable, microfluidic platform capable of simultaneous, multiplexed detection of several of the bacterial pathogens that cause these diseases. This platform can perform fast, sensitive immunoassays directly from relevant, complex clinical matrices such as stool without extensive sample cleanup or preparation. Using only 1 µL of sample per assay, we demonstrate simultaneous multiplexed detection of four bacterial pathogens implicated in diarrheal and enteric diseases in less than 20 min. Full article