Biosensors — Open Access Journal

Freestyle Libre (FL) is a factory calibrated Flash Glucose Monitor (FGM). We investigated Mean Absolute Relative Difference (MARD) between Self Monitoring of Blood Glucose (SMBG) and FL measurements in the first day of sensor wear in 39 subjects with Type 1 diabetes. The overall MARD was 12.3%, while the individual MARDs ranged from 4% to 25%. Five participants had a MARD ≥ 20%. We estimated bias and lag between the FL and SMBG measurements. The estimated biases range from −1.8 $\mathrm{mmol}$/$\mathrm{L}$ to 1.4 $\mathrm{mmol}$/$\mathrm{L}$, and lags range from 2 $\min$ to 24 $\min$. Bias is identified as a main cause of poor individual MARDs. The biases seem to persist in days 2–7 of sensor usage. All cases of MARD ≥ 20% in the first day are eliminated by bias correction, and overall MARD is reduced from 12.3% to 9.2%, indicating that adding support for voluntary user-supplied bias correction in the FL could improve its performance. Full article

This review focuses on the fabrication of biosensors using metal-organic frameworks (MOFs) as recognition and/or transducer elements. A brief introduction discussing the importance of the development of new biosensor schemes is presented, describing these coordination polymers, their properties, applications, and the main advantages and drawbacks for the final goal. The increasing number of publications regarding the characteristics of these materials and the new micro- and nanofabrication techniques allowing the preparation of more accurate, robust, and sensitive biosensors are also discussed. This work aims to offer a new perspective from the point of view of materials science compared to other reviews focusing on the transduction mechanism or the nature of the analyte. A few examples are discussed depending on the starting materials, the integration of the MOF as a part of the biosensor and, in a deep detail, the fabrication procedure. Full article

The design of new materials as active layers is important for electrochemical sensor and biosensor development. Among the techniques for the modification and functionalization of electrodes, the laser induced forward transfer (LIFT) has emerged as a powerful physisorption method for the deposition of various materials (even labile materials like enzymes) that results in intimate and stable contact with target surface. In this work, Pt, Au, and glassy carbon screen printed electrodes (SPEs) treated by LIFT with phosphate buffer have been characterized by scanning electron microscopy and atomic force microscopy to reveal a flattening effect of all surfaces. The electrochemical characterization by cyclic voltammetry shows significant differences depending on the electrode material. The electroactivity of Au is reduced while that of glassy carbon and Pt is greatly enhanced. In particular, the electrochemical behavior of a phosphate LIFT treated Pt showed a marked enrichment of hydrogen adsorbed layer, suggesting an elevated electrocatalytic activity towards glucose oxidation. When Pt electrodes modified in this way were used as an effective glucose sensor, a 1–10 mM linear response and a 10 µM detection limit were obtained. A possible role of phosphate that was securely immobilized on a Pt surface, as evidenced by XPS analysis, enhancing the glucose electrooxidation is discussed. Full article

Electric Cell-Substrate Impedance Sensing (ECIS) can produce reproducible wounding models by mechanically disrupting a cell monolayer. This study compared in vitro wound-healing using human cerebral microvascular endothelial cells (hCMVEC) with both single electrode (8W1E) and multiple electrodes (8W10E+) arrays. Measurements of hCMVEC migration and barrier functions were conducted, revealing variable levels of barrier disruption could be achieved by altering the duration and magnitude of the applied current. In all scenarios, the barrier (Rb) did not recover the strength observed prior to injury. Localization of junctional proteins following wounding were analyzed by immunocytochemistry. Following wounding, cell migration was generally faster on the 8W10E+ than the 8W1E array. Immunohistochemical analysis revealed non-viable cells remained on the 8W1E electrodes but not the 8W10E+ electrodes. However, viable cells partially remained on the 8W10E+ electrodes following wounding. In addition, the 8W10E+ electrodes demonstrated variation in cell loss across electrodes within the same well. This suggests the type of wounding is different on the two array types. However, our data show both arrays can be used to model incomplete barrier recovery and therefore both have potential for testing of drugs to improve endothelial barrier function. This is the first time that the possibility of using the 8W10E+ array as a wounding model is addressed. We highlight the differences in wounding produced between the two arrays, and can be used to study the underlying causes for impaired barrier function following CNS injuries. Full article

Förster resonance energy transfer (FRET) between fluorophores of the same species was recognized in the early to mid-1900s, well before modern heterotransfer applications. Recently, homotransfer FRET principles have re-emerged in biosensors that incorporate genetically encoded fluorescent proteins. Homotransfer offers distinct advantages over the standard heterotransfer FRET method, some of which are related to the use of fluorescence polarization microscopy to quantify FRET between two fluorophores of identical color. These include enhanced signal-to-noise, greater compatibility with other optical sensors and modulators, and new design strategies based upon the clustering or dimerization of singly-labeled sensors. Here, we discuss the theoretical basis for measuring homotransfer using polarization microscopy, procedures for data collection and processing, and we review the existing genetically-encoded homotransfer biosensors. Full article

Adverse cardiac events are a major cause of late-stage drug development withdrawals. Improved in vitro systems for predicting cardiotoxicity are of great interest to prevent these events and to reduce the expenses involved in the introduction of cardiac drugs into the marketplace. Interdigitated electrodes (IDEs) affixed with a culture well provide a simple, suitable solution for in vitro analysis of cells because of their high sensitivity, ease of fabrication, and label-free, nondestructive analysis. Culturing human pluripotent stem cell differentiated cardiomyocytes onto these IDEs allows for the use of the IDE–cell combination in predictive toxicity assays. IDEs with smaller interdigitated distances allow for greater sensitivity, but typically require cleanroom fabrication. In this communication, we report the definition of a simple IDE geometry on a printed nanostructured substrate, demonstrate a Cellular Index (CI) increase from 0 to 7.7 for human cardiomyocytes, and a decrease in CI from 2.3 to 1 with increased concentration of the model drug, norepinephrine. The nanostructuring results in an increased sensitivity of our 1 mm pitch IDEs when compared to traditionally fabricated IDEs with a pitch of 10 μm (100 times larger electrode gap). The entire nanostructured IDE (nIDE) is fabricated and assembled in a rapid nanofabrication environment, thus allowing for iterative design changes and robust fabrication of devices. Full article

In the present contribution the fabrication and characterization of functionalized gold nanospheres of uniform shape and controlled size is reported. These nano-objects are intended to be used as Surface Enhanced Raman Spectroscopy (SERS) sensors for in-vitro cellular uptake and localization. Thiophenol was used as molecular reporter and was bound to the Au surface by a chemisorption process in aqueous solution. The obtained colloidal solution was highly stable and no aggregation of the single nanospheres into larger clusters was observed. The nanoparticles were incubated in human prostatic cells with the aim of developing a robust, SERS-based method to differentiate normal and tumor cell lines. SERS imaging experiments showed that tumor cells uptake considerably larger amounts of nanoparticles in comparison to normal cells (up to 950% more); significant differences were also observed in the uptake kinetics. This largely different behaviour might be exploited in diagnostic and therapeutic applications. Full article

Cardiovascular diseases, including atherosclerosis, now account for more deaths in the Western world than from any other cause. Atherosclerosis has a chronic inflammatory component involving Th1 pro-inflammatory cytokines such as IFN-γ, which is known to induce endothelial cell inflammatory responses. On the other hand CNP, which acts via its receptors to elevate intracellular cGMP, is produced by endothelium and endocardium and is upregulated in atherosclerosis. It is believed to be protective, however its role in vascular inflammation is not well understood. The aim of this study was to investigate the effects of CNP on human endothelial cell inflammatory responses following IFN-γ stimulation. Human umbilical vein endothelial cells were treated with either IFN-γ (10 ng/mL) or CNP (100 nm), or both in combination, followed by analysis by flow cytometry for expression of MHC class I and ICAM-1. IFN-γ significantly increased expression of both molecules, which was significantly inhibited by CNP or the cGMP donor 8-Bromoguanosine 3’,5’-cyclic monophosphate (1 µm). CNP also reduced IFN-γ mediated kynurenine generation by the IFN-γ regulated enzyme indoleamine-2,3-deoxygenase (IDO). We conclude that CNP downmodulates IFN-γ induced pro-inflammatory gene expression in human endothelial cells via a cGMP-mediated pathway. Thus, CNP may have a protective role in vascular inflammation and novel therapeutic strategies for CVD based on upregulation of endothelial CNP expression could reduce chronic EC inflammation. Full article

An automatic spot identification method is developed for high throughput surface plasmon resonance imaging (SPRi) analysis. As a combination of video accessing, image enhancement, image processing and parallel processing techniques, the method can identify the spots in SPRi images of the microarray from SPRi video data. In demonstrations of the method, SPRi video data of different protein microarrays were processed by the method. Results show that our method can locate spots in the microarray accurately regardless of the microarray pattern, spot-background contrast, light nonuniformity and spotting defects, but also can provide address information of the spots. Full article

In this study, the electropolymerization of 4-hydroxyphenylacetic acid (4-HPA) over graphite electrodes (GE) was optimized, aiming its application as a functionalized electrochemical platform for oligonucleotides immobilization. It was investigated for the number of potential cycles and the scan rate influence on the monomer electropolymerization by using cyclic voltammetry technique. It was observed that the polymeric film showed a redox response in the region of +0.53/+0.38 V and the increase in the number of cycles produces more electroactive platforms because of the better electrode coverage. On the other hand, the decrease of scan rate produces more electroactive platforms because of the occurrence of more organized coupling. Scanning electron microscopy (SEM) images showed that the number of potential cycles influences the coverage and morphology of the electrodeposited polymeric film. However, the images also showed that at different scan rates a more organized material was produced. The influence of these optimized polymerization parameters was evaluated both in the immobilization of specific oligonucleotides and in the detection of hybridization with complementary target. Poly(4-HPA)/GE platform has shown efficient and sensitive for oligonucleotides immobilization, as well as for a hybridization event with the complementary oligonucleotide in all investigated cases. The electrode was modified with 100 cycles at 75 mV/s presented the best responses in function of the amplitude at the monitored peak current values for the Methylene Blue and Ethidium Bromide intercalators. The construction of the genosensor to detect a specific oligonucleotide sequence for the Mycobacterium tuberculosis bacillus confirmed the results regarding the poly(4-HPA)/GE platform efficiency since it showed excellent sensitivity. The limit of detection and the limit of quantification was found to be 0.56 (±0.05) μM and 8.6 (±0.7) μM, respectively operating with very low solution volumes (15 µL of probe and 10 µL target). The biosensor development was possible with optimization of the probe adsorption parameters and target hybridization, which led to an improvement in the decrease of the Methylene Blue (MB) reduction signal from 14% to 34%. In addition, interference studies showed that the genosensor has satisfactory selectivity since the hybridization with a non-specific probe resulted in a signal decrease (46% lower) when compared to the specific target. Full article

In order to exploit plants as environmental biosensors, previous researches have been focused on the electrical signal response of the plants to different environmental stimuli. One of the important outcomes of those researches has been the extraction of meaningful features from the electrical signals and the use of such features for the classification of the stimuli which affected the plants. The classification results are dependent on the classifier algorithm used, features extracted and the quality of data. This paper presents an innovative way of extracting features from raw plant electrical signal response to classify the external stimuli which caused the plant to produce such a signal. A curve fitting approach in extracting features from the raw signal for classification of the applied stimuli has been adopted in this work, thereby evaluating whether the shape of the raw signal is dependent on the stimuli applied. Four types of curve fitting models—Polynomial, Gaussian, Fourier and Exponential, have been explored. The fitting accuracy (i.e., fitting of curve to the actual raw signal) depicted through R-squared values has allowed exploration of which curve fitting model performs best. The coefficients of the curve fit models were then used as features. Thereafter, using simple classification algorithms such as Linear Discriminant Analysis (LDA), Quadratic Discriminant Analysis (QDA) etc. within the curve fit coefficient space, we have verified that within the available data, above 90% classification accuracy can be achieved. The successful hypothesis taken in this work will allow further research in implementing plants as environmental biosensors. Full article

In this work, we developed a novel fluorescent sensor by combining molecularly imprinted polymers (MIPs) with graphene quantum dots (GQDs) for the determination of tetracycline (TC) in aqueous samples. Firstly, we developed a one-pot green method to synthesize GQDs as the fluorescent probes. GQDs with carboxyl groups or amino groups were fabricated. It was found that carboxyl groups played an important role in the fluorescence quenching. Based on these findings, the GQDs-MIPs microspheres were prepared using a sol-gel process. GQDs-MIPs showed strong fluorescent emission at 410 nm when excited at 360 nm, and the fluorescence was quenched in the presence of TC. Under optimum conditions, the fluorescence intensity of GQDs-MIPs decreased in response to the increase of TC concentration. The linear rage was from 1.0 to 10⁴ µg·L⁻¹, and the limit of detection was determined to be 1 µg·L⁻¹. The GQDs-MIPs also demonstrated high selectivity towards TC. The fluorescent sensor was successfully applied for the detection of TC in real spiked milk samples. Full article

Colorimetric biosensors of cholinesterase inhibitors are ideal for fast, reliable, and very simple detection of agents in air, in water, and on surfaces. This paper describes an innovation of the Czech Detehit biosensor, which is based on a biochemical enzymatic reaction visualized by using Ellman’s reagent as a chromogenic indicator. The modification basically consists of a much more distinct color response of the biosensor, attained through optimization of the reaction system by using Guinea Green B as the indicator. The performance of the modified biosensor was verified on the chemical warfare agents (sarin, soman, cyclosarin, and VX) in water. The detection limits ascertained visually (with the naked eye) were about 0.001 µg/mL in water (exposure time 60 s, inhibition efficiency 25%). Full article

The notion of surface plasmon resonance (SPR) sensor research emerged more than eight decades ago from the first observed phenomena in 1902 until the first introduced principles for gas sensing and biosensing in 1983. The sensing platform has been hand-in-hand with the plethora of sensing technology advancement including nanostructuring, optical technology, fluidic technology, and light source technology, which contribute to substantial progress in SPR sensor evolution. Nevertheless, the commercial products of SPR sensors in the market still require high-cost investment, component, and operation, leading to unaffordability for their implementation in a low-cost point of care (PoC) or laboratories. In this article, we present a comprehensive review of SPR sensor development including the state of the art from a perspective of light source technology trends. Based on our review, the trend of SPR sensor configurations, as well as its methodology and optical designs are strongly influenced by the development of light source technology as a critical component. These simultaneously offer new underlying principles of SPR sensor towards miniaturization, portability, and disposability features. The low-cost solid-state light source technology, such as laser diode, light-emitting diode (LED), organic light emitting diode (OLED) and smartphone display have been reported as proof of concept for the future of low-cost SPR sensor platforms. Finally, this review provides a comprehensive overview, particularly for SPR sensor designers, including emerging engineers or experts in this field. Full article

The focus of this study is to design and integrate silver/silver chloride (Ag/AgCl) electronic textile (e-textile) electrodes into different textile substrates to evaluate their ability to monitor electrodermal activity (EDA). Ag/AgCl e-textiles were stitched into woven textiles of cotton, nylon, and polyester to function as EDA monitoring electrodes. EDA stimulus responses detected by dry e-textile electrodes at various locations on the hand were compared to the EDA signals collected by dry solid Ag/AgCl electrodes. 4-h EDA data with e-textile and clinically conventional rigid electrodes were compared in relation to skin surface temperature. The woven cotton textile substrate with e-textile electrodes (0.12 cm² surface area, 0.40 cm distance) was the optimal material to detect the EDA stimulus responses with the highest average Pearson correlation coefficient of 0.913 ± 0.041 when placed on the distal phalanx of the middle finger. In addition, differences with EDA waveforms recorded on various fingers were observed. Trends of long-term measurements showed that skin surface temperature affected EDA signals recorded by non-breathable electrodes more than when e-textile electrodes were used. The effective design criteria outlined for e-textile electrodes can promote the development of comfortable and unobtrusive EDA monitoring systems, which can help improve our knowledge of the human neurological system. Full article

The viscosity variation of sputum is a common symptom of the progression of Chronic Obstructive Pulmonary Disease (COPD). Since the hydration of the sputum defines its viscosity level, dielectric sensors could be used for the characterization of sputum samples collected from patients for early diagnosis of COPD. In this work, a CMOS-based dielectric sensor for the real-time monitoring of sputum viscosity was designed and fabricated. A proper packaging for the ESD-protection and short-circuit prevention of the sensor was developed. The performance evaluation results show that the radio frequency sensor is capable of measuring dielectric constant of biofluids with an accuracy of 4.17%. Integration of this sensor into a portable system will result in a hand-held device capable of measuring viscosity of sputum samples of COPD-patients for diagnostic purposes. Full article

The present study was aimed to develop “fluorine doped” tin oxide glass electrode with a MoSe₂ nano-urchin based electrochemical biosensor for detection of Escherichia coli Shiga toxin DNA. The study comprises two conductive electrodes, and the working electrodes were drop deposited using MoSe₂ nano-urchin, and DNA sequences specific to Shiga toxin Escherichia coli. Morphological characterizations were performed using Fourier transforms infrared spectrophotometer; X-ray diffraction technique and scanning electron microscopy. All measurements were done using methylene blue as an electrochemical indicator. The proposed electrochemical geno-sensor showed good linear detection range of 1 fM–100 µM with a low detection limit of 1 fM where the current response increased linearly with Escherichia coli Shiga toxin dsDNA concentration with R² = 0.99. Additionally, the real sample was spiked with the dsDNA that shows insignificant interference. The results revealed that the developed sensing platform significantly improved the sensitivity and can provide a promising platform for effective detection of biomolecules using minute samples due to its stability and sensitivity. Full article

In red blood cell (RBC) disorders, such as sickle cell disease, hereditary spherocytosis, and diabetes, alterations to the size and shape of RBCs due to either mutations of RBC proteins or changes to the extracellular environment, lead to compromised cell deformability, impaired cell stability, and increased propensity to aggregate. Numerous laboratory approaches have been implemented to elucidate the pathogenesis of RBC disorders. Concurrently, computational RBC models have been developed to simulate the dynamics of RBCs under physiological and pathological conditions. In this work, we review recent laboratory and computational studies of disordered RBCs. Distinguished from previous reviews, we emphasize how experimental techniques and computational modeling can be synergically integrated to improve the understanding of the pathophysiology of hematological disorders. Full article

A theoretical analysis and computational study of biomaterial sample detection with surface plasmon resonance (SPR) phenomenon spectroscopy are presented in this work with the objective of achieving more sensitive detection. In this paper, a Fe₃O₄@Au core-shell, a nanocomposite spherical nanoparticle consisting of a spherical Fe₃O₄ core covered by an Au shell, was used as an active material for biomaterial sample detection, such as for blood plasma, haemoglobin (Hb) cytoplasm and lecithin, with a wavelength of 632.8 nm. We present the detection amplification technique through an attenuated total reflection (ATR) spectrum in the Kretschmann configuration. The system consists of a four-layer material, i.e., prism/Ag/Fe₃O₄@Au + biomaterial sample/air. The effective permittivity determination of the core-shell nanoparticle (Fe₃O₄@Au) and the composite (Fe₃O₄@Au + biomaterial sample) was done by applying the effective medium theory approximation, and the calculation of the reflectivity was carried out by varying the size of the core-shell, volume fraction and biomaterial sample. In this model, the refractive index (RI) of the BK7 prism is 1.51; the RI of the Ag thin film is 0.13455 + 3.98651i with a thickness of 40 nm; and the RI of the composite is varied depending on the size of the nanoparticle core-shell and the RI of the biomaterial samples. Our results show that by varying the sizes of the core-shell, volume fraction and the RIs of the biomaterial samples, the dip in the reflectivity (ATR) spectrum is shifted to the larger angle of incident light, and the addition of a core-shell in the conventional SPR-based biosensor leads to the enhancement of the SPR biosensor sensitivity. For a core-shell with a radius a = 2.5 nm, the sensitivity increased by 10% for blood plasma detection, 47.72% for Hb cytoplasm detection and by 22.08% for lecithin detection compared to the sensitivity of the conventional SPR-based biosensor without core-shell addition. Full article