Exosomes secreted from cells carry rich information from their parent cells, representing a promising biomarker for investigation of diseases. We develop a dual-nanopore biosensor using DNA aptamers to specifically recognize CD63 protein on the exosome’s surface, which enables label-free exosome detection based on ionic current change. The sensor allows for sensitive detection of exosomes with a detection limit of 3.4 × 106 particles/mL. The dual-nanopore biosensor was able to form an intrapipette electric circuit for ionic current measurement due to its unique structure, which is crucial to achieve detection of exosome secretion from a single cell. We utilized a microwell array chip to entrap a single cell into a confined microwell with small volume, enabling the accumulation of exosomes with high concentration. The dual-nanopore biosensor was positioned into the microwell with a single cell, and monitoring of exosome secretion from a single cell in different cell lines and under different stimulations has been achieved. Our design may provide a useful platform for developing nanopore biosensors for detecting cell secretions from a single living cell.

Core body temperature (CBT) is one of the four vital signs that must be monitored continuously. The continuous recording of CBT is possible through invasive methods by inserting a temperature probe into specific body sites. We report a novel method to monitor CBT through the quantitative measurement of skin blood perfusion rate (ωb,skin). By monitoring the skin temperature, heat flux, and ωb,skin, the arterial blood temperature, equivalent to CBT, can be extracted. ωb,skin is quantitatively evaluated thermally via sinusoidal heating with regulated thermal penetration depth so that the blood perfusion rate is acquired only in the skin. Its quantification is significant because it indicates various physiological events including hyper- or hypothermia, tissue death, and delineation of tumors. A subject showed promising results with steady values of ωb,skin and CBT of 5.2 ± 1.05 × 10–4 s–1 and 36.51 ± 0.23 °C, respectively. For periods where the subject’s actual CBT (axillary temperature) did not fall within the estimated range, the average deviation from the actual CBT was only 0.07 °C. This study aims to develop a competent methodology capable of continuously monitoring the CBT and blood perfusion rate at a distant location from the core body region for the diagnosis of a patient’s health condition with wearable devices.

Coronary heart disease (CHD) poses an important threat to human health, and its pathogenesis is the formation of atheromatous plaques in coronary ventricles. Compared to other biomarkers, lipoprotein-associated phospholipase A2 (Lp-PLA2), which is involved in multiple processes of atherosclerosis, is a noticeable inflammatory biomarker related to CHD. Herein, using a multifunctional nanocomposite containing a CoFe Prussian blue analogue (PBA) and Au nanoparticles (AuNPs) (AuNPs@CoFe PBA) as a sensing substrate, an electrochemiluminescent (ECL) immunosensor was developed for the highly sensitive detection of Lp-PLA2. Benefiting from the synergistic effect of the PBA and AuNPs, the nanocomposite exhibits excellent peroxidase-like activity and can catalyze the luminol–ECL reaction, amplifying the ECL signal by ∼29-fold. Meanwhile, the enlarged specific surface area of the nanocomposite and the presence of abundant AuNPs allow the immobilization of more antibody proteins, thereby improving the sensing response of the immunosensor. When the target Lp-PLA2 is captured by the antibody on the sensor surface, the sensor emits a reduced ECL signal because of the increased mass and electron transfer resistance due to the formation of the immune complex. Under optimized conditions, the constructed ECL immunosensor exhibits a broad linear range from 1 to 2200 ng/mL and a low detection limit of 0.21 ng/mL. Additionally, the ECL immunosensor exhibits high specificity, stability, and reproducibility. This work provides a new approach to diagnose CHD and broadened the application of the PBA in the field of ECL sensors.

Potentiometric ion-selective electrodes (ISEs) have broad applications in personalized healthcare, smart agriculture, oil/gas exploration, and environmental monitoring. However, high-precision potentiometric sensing is difficult with field-deployed sensors due to time-dependent voltage drift and the need for frequent calibration. In the laboratory setting, these issues are resolved by repeated calibration by measuring the voltage response at multiple standard solutions at a constant temperature. For field-deployed sensors, it is difficult to frequently interrupt operation and recalibrate with standard solutions. Moreover, the constant surrounding temperature constraint imposed by the traditional calibration process makes it unsuitable for temperature-varying field use. To address the challenges of traditional calibration for field-deployed sensors, in this study, we propose a novel in situ calibration approach in which we use natural/external temperature variation in the field to obtain the time-varying calibration parameters, without having to relocate the sensors or use any complex system. We also develop a temperature-supervised monitoring method to detect the drift of the sensor during operation. Collectively, the temperature-based drift monitoring and in situ calibration methods allow us to monitor the drift of sensors and correct them periodically to achieve high-precision sensing. We demonstrate our approach in three testbeds: (1) under controlled temperature variation in the lab, (2) under natural temperature variation in a greenhouse, and (3) in the field to monitor nitrate activity of an agricultural site. In the laboratory study, we validate that the calibration parameters of printed nitrate ISEs can be reproduced by our proposed calibration process; therefore, it can serve as an alternative to traditional calibration processes. In the greenhouse, we show the use of natural temperature variation to calibrate the sensors and detect the drift in a fixed concentration nitrate solution. Finally, we demonstrate the use of the method to monitor the nitrate activity of an agricultural field within 10% of laboratory-based measurements (i.e., a sensitivity of 0.03 mM) for a period of 22 days. The findings highlight the prospect of temperature-based calibration and drift monitoring for high-precision sensing with field-deployed ISEs.

We demonstrate the reliable creation of multiple layers of Au nanoparticles in random close-packed arrays with sub-nm gaps as a sensitive surface-enhanced Raman scattering substrate. Using oxygen plasma etching, all the original molecules creating the nanogaps can be removed and replaced with scaffolding ligands that deliver extremely consistent gap sizes below 1 nm. This allows precision tailoring of the chemical environment of the nanogaps which is crucial for practical Raman sensing applications. Because the resulting aggregate layers are easily accessible from opposite sides by fluids and by light, high-performance fluidic sensing cells are enabled. The ability to cyclically clean off analytes and reuse these films is shown, exemplified by sensing of toluene, volatile organic compounds, and paracetamol, among others.

Bioelectronic tongues based on umami taste receptors have recently been reported for versatile applications such as food analyses. However, their practical applications are still limited, partly due to their limited stability and non-specific responses in real sample environments. Herein, we have developed a hydrogel-based bioelectronic tongue for the sensitive assessment of umami intensity in fish extract samples. In this study, the T1R1 venus flytrap of an umami taste receptor was immobilized on the gold floating electrodes of a carbon nanotube-based field-effect transistor. A polyacrylamide conducting hydrogel film was further hybridized on the sensor surface via physical adsorption, which could provide a good physiological environment to maintain the activity of receptors due to its excellent hydrophilicity and biocompatibility. The bioelectronic tongue with a receptor-embedded hydrogel structure showed a sensitive detection of umami substances down to 1 fM, and it also had a wide detection range of 10–15–10–2 M for monosodium glutamate and disodium inosinate, which covers the human taste threshold. More importantly, the proposed sensor could significantly reduce the non-specific binding of non-target molecules to a carbon nanotube channel as well as exhibit long-term stability, enabling sensitive detection of umami substances even in fish extract samples. Our hydrogel-based bioelectronic tongue provides a promising platform for future applications such as the flavor evaluation of foods and beverages.

Hydrocarbon (HC) monitoring is necessary for safe and effective operations in industries such as petroleum and gas. In this study, total hydrocarbons can be detected by using yttria-stabilized zirconia (YSZ)-based potentiometric-type gas sensor using MgFe2O4 sensing electrode (SE). The sensor was found to generate a similar response magnitude to those of hydrocarbons that have the same carbon number, irrespective of the type of carbon bond (total hydrocarbon detection). Aside from being capable of detecting total hydrocarbons sensitively and selectively with rapid response time, the sensor using MgFe2O4-SE also exhibited a linear relationship between sensor responses and carbon number. In addition to that, the developed sensor showed a logarithmically linear relationship between sensor responses and HC concentration in the range 20–700 ppm. These sensing characteristics were confirmed to be reproducible, and sensor responses toward HC were found to be repeatable and gradually decreased with increasing in O2 concentration in the range of 3–21 vol %.

The selectivity of the linear polymer chain toward its binding moieties has been considered negligible; thus, a clear demonstration showing the best-fit binding of a linear polymer to its guest counterpart is still unknown. Luminescent poly(acrylic acid) (PAA)-stabilized silver nanodots (PAA-AgNDs) have been applied as a turn-on sensor to monitor the interaction between the PAA chain and its binding cations. The binding of cations ions to the PAA chain may cross-link the linear PAA chain via coordination with carboxylate, which increases the rigidity of the polymer chain, retards the nonradiative decay of PAA-AgNDs, and consequently enhances the emission of silver nanodots while inducing a blue-shift of its emission spectrum. For the first time, we have demonstrated that a linear polymer chain can act as an open host to selectively bind to its best-matching cations. Specifically, among Group 2 cations (Mg2+, Ca2+, Sr2+, Ba2+), calcium ions show the strongest bonding to the PAA polymer chain. Our research suggests that, with extra rigidity, the polymer improves its chemical stability as calcium ions cross-linked the linear polymer. Meanwhile, it has also been demonstrated that luminescent silver nanodots can be excellent probes for the detection of polymer activities with straightforward and simple visualization methods.

Flexible, water-soluble hosts are capable of selective molecular recognition in cellular environments and can detect neurotransmitters such as choline in cells. Both cationic and anionic water-soluble self-folded deep cavitands can recognize suitable styrylpyridinium dyes in cellular interiors. The dyes selectively accumulate in nucleotide-rich regions of the cell nucleus and cytoplasm. The hosts bind the dyes and promote their relocation to the outer cell membrane: the lipophilic cavitands predominantly reside in membrane environments but are still capable of binding suitable targets in other cellular organelles. Incubating the cells with structurally similar biomarkers such as choline, cholamine, betaine, or butyrylcholine illustrates the selective recognition. Choline and butyrylcholine can be bound by the hosts, but minimal binding is seen with betaine or cholamine. Varying the dye allows control of the optical detection method, and both “turn-on” sensing and “turn-off” sensing are possible.

Health initiatives worldwide demand affordable point-of-care devices to aid in the reduction of morbidity and mortality rates of high-incidence infectious and noncommunicable diseases. However, the production of robust and reliable easy-to-use diagnostic platforms showing the ability to quantitatively measure several biomarkers in physiological fluids and that could in turn be decentralized to reach any relevant environment remains a challenge. Here, we show the particular combination of paper-microfluidic technology, electrochemical transduction, and magnetic nanoparticle-based immunoassay approaches to produce a unique, compact, and easily deployable multiplex device to simultaneously measure interleukin-8, tumor necrosis factor-α, and myeloperoxidase biomarkers in sputum, developed with the aim of facilitating the timely detection of acute exacerbations of chronic obstructive pulmonary disease. The device incorporates an on-chip electrochemical cell array and a multichannel paper component, engineered to be easily aligned into a polymeric cartridge and exchanged if necessary. Calibration curves at clinically relevant biomarker concentration ranges are produced in buffer and artificial sputum. The analysis of sputum samples of healthy individuals and acutely exacerbated patients produces statistically significant biomarker concentration differences between the two studied groups. The device can be mass-produced at a low cost, being an easily adaptable platform for measuring other disease-related target biomarkers.

Antibodies are among the most relevant biomolecular targets for diagnostic and clinical applications. In this Perspective, we provide a critical overview of recent research efforts focused on the development and characterization of devices, switches, and reactions based on the use of synthetic antigen-conjugated DNA strands designed to be responsive to specific antibodies. These systems can find applications in sensing, drug-delivery, and antibody–antigen binding characterization. The examples described here demonstrate how the programmability and chemical versatility of synthetic nucleic acids can be used to create innovative analytical tools and target-responsive systems with promising potentials.

Although there is a growing demand for new sensors for environmental monitoring, biofouling continues to plague current sensors and sensing networks. As soon as a sensor is placed in water, the formation of a biofilm begins. Once a biofilm is established, reliable measurements are often no longer possible. Although current biofouling mitigation strategies can slow the biofouling process, a biofilm will eventually develop on or near the sensing surface. While antibiofouling strategies are being continuously developed, the complexity of the biofilm community structure and the surrounding environment means that there is unlikely to be a single solution that will minimize biofilms on all environmental sensors. Thus, antibiofouling research often focuses on optimizing a specific biofilm mitigation approach for a given sensor, application, and environmental condition. While this is practical from the standpoint of a sensor developer, it makes the comparison of different mitigation strategies difficult. In this Perspective, we discuss the application of different biofouling mitigation strategies to sensing and then explore the need for the sensor community to adopt standard protocols to increase the comparability of the biofouling mitigation approaches and help sensor developers identify the most appropriate strategy for their system.

Electrochemical detection methods are attractive for developing miniaturized, disposable, and portable sensors for molecular diagnostics. In this article, we present a cucurbit[7]uril-based chemosensor with an electrochemical signal readout for the micromolar detection of the muscle relaxant pancuronium bromide in buffer and human urine. This is possible through a competitive binding assay using a chemosensor ensemble consisting of cucurbit[7]uril as the host and an electrochemically active platinum(II) compound as the guest indicator. The electrochemical properties of the indicator are strongly modulated depending on the complexation state, a feature that is exploited to establish a functional chemosensor. Our design avoids cumbersome immobilization approaches on electrode surfaces, which are associated with practical and conceptual drawbacks. Moreover, it can be used with commercially available screen-printed electrodes that require minimal sample volume. The design principle presented here can be applied to other cucurbit[n]uril-based chemosensors, providing an alternative to fluorescence-based assays.

Human papillomavirus (HPV) is the causative agent for cervical cancer. Of the various types of HPV, the high-risk HPV-16 type is the most important antigenic high-risk HPV. In this work, the antigenic HPV-16 L1 peptide was immobilized on a glassy carbon electrode and used to detect several concentrations of the anti-HPV-16 L1 antibody, and vice versa. Two electrode platforms were used: onion-like carbon (OLC) and its polyacrylonitrile (OLC-PAN) composites. Both platforms gave a wide linear concentration range (1.95 fg/mL to 6.25 ng/mL), excellent sensitivity (>5.2 μA/log ([HPV-16 L1, fg/mL]), and extra-ordinarily low limit of detection (LoD) of 1.83 fg/mL (32.7 aM) and 0.61 fg/mL (10.9 aM) for OLC-PAN and OLC-based immunosensors, respectively. OLC-PAN modified with the HPV-16 L1 protein showed low LoD for the HPV-16 L1 antibody (2.54 fg/mL, i.e., 45.36 aM), proving its potential use for screening purposes. The specificity of detection was proven with the anti-ovalbumin antibody (anti-OVA) and native ovalbumin protein (OVA). An immobilized antigenic HPV-16 L1 peptide showed insignificant interaction with anti-OVA in contrast with the excellent interaction with anti-HPV-16 L1 antibody, thus proving high specificity. The application of the immunosensor as a potential point-of-care (PoC) diagnostic device was investigated with screen-printed carbon electrodes, which detected ultra-low (ca. 0.7 fg/mL ≈ 12.5 aM) and high (ca. 12 μg/mL ≈ 0.21 μM) concentrations. This study represents the lowest LoD reported for HPV-16 L1. It opens the door for further investigation with other electrode platforms and realization of PoC diagnostic devices for screening and testing of HPV biomarkers for cervical cancer.

Their chemical diversity, uniform pore sizes, and large internal surface areas make metal–organic frameworks (MOFs) highly suitable for volatile organic compound (VOC) adsorption. This work compares two geometries of capacitive VOC sensors that use the MOF material ZIF-8 as an affinity layer. When using a permeable top electrode (thickness < 25 nm), the metal–insulator–metal (MIM) sandwich configuration exhibits superior sensitivity, an improved detection limit, and a smaller footprint than the conventional interdigitated electrode layout. Moreover, the transduction of VOC adsorption in ZIF-8 via MIM capacitors is more sensitive to polar VOCs and provides better selectivity at high loadings than gravimetric and optical transductions.

Serving as the interface between fetal and maternal circulation, the placenta plays a critical role in fetal growth and development. Placental exosomes are small membrane-bound extracellular vesicles released by the placenta during pregnancy. They contain a variety of biomolecules, including lipids, proteins, and nucleic acids, which can potentially be biomarkers of maternal diseases. An increasing number of studies have demonstrated the utility of placental exosomes for the diagnosis and monitoring of pathological conditions such as pre-eclampsia and gestational diabetes. This suggests that placental exosomes may serve as new biomarkers in liquid biopsy analysis. This review provides an overview of the current understanding of the biological function of placental exosomes and their potential as biomarkers of maternal diseases. Additionally, this review highlights current barriers and the way forward for standardization and validation of known techniques for exosome isolation, characterization, and detection. Finally, microfluidic devices for exosome research are discussed.

Airborne transmission via virus-laden aerosols is a dominant route for the transmission of respiratory diseases, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Direct, non-invasive screening of respiratory virus aerosols in patients has been a long-standing technical challenge. Here, we introduce a point-of-care testing platform that directly detects SARS-CoV-2 aerosols in as little as two exhaled breaths of patients and provides results in under 60 s. It integrates a hand-held breath aerosol collector and a llama-derived, SARS-CoV-2 spike-protein specific nanobody bound to an ultrasensitive micro-immunoelectrode biosensor, which detects the oxidation of tyrosine amino acids present in SARS-CoV-2 viral particles. Laboratory and clinical trial results were within 20% of those obtained using standard testing methods. Importantly, the electrochemical biosensor directly detects the virus itself, as opposed to a surrogate or signature of the virus, and is sensitive to as little as 10 viral particles in a sample. Our platform holds the potential to be adapted for multiplexed detection of different respiratory viruses. It provides a rapid and non-invasive alternative to conventional viral diagnostics.

Urinary tract infection (UTI) diagnosis based on urine culture for bacteriuria analysis is time-consuming and often leads to wastage of hospital resources due to false-positive UTI cases. Direct cellular phenotyping (e.g., RBCs, neutrophils, epithelial cells) of urine samples remains a technical challenge as low cell concentrations, and urine characteristics (conductivities, pH, microbes) can affect the accuracy of cell measurements. In this work, we report a microfluidic inertial-impedance cytometry technique for label-free rapid (<5 min) neutrophil sorting and impedance profiling from urine directly. Based on size-based inertial focusing effects, neutrophils are isolated, concentrated, and resuspended in saline (buffer exchange) to improve consistency in impedance-based single-cell analysis. We first observed that both urine pH and the presence of bacteria can affect neutrophil high-frequency impedance measurements possibly due to changes in nucleus morphology as neutrophils undergo NETosis and phagocytosis, respectively. As a proof-of-concept for clinical testing, we report for the first time, rapid UTI testing based on multiparametric impedance profiling of putative neutrophils (electrical size, membrane properties, and distribution) in urine samples from non-UTI (n = 20) and UTI patients (n = 20). A significant increase in cell count was observed in UTI samples, and biophysical parameters were used to develop a UTI classifier with an area under the receiver operating characteristic curve of 0.84. Overall, the developed platform facilitates rapid culture-free urine screening which can be further developed to assess disease severity in UTI and other urologic diseases based on neutrophil electrical signatures.

Although continuous monitoring of constituents in complex sweat is crucial for noninvasive physiological evaluation, biofouling on the sweat sensor surface and inadequate flexible self-healing materials restrict its applications. Herein, a fully self-healing and strong anti-biofouling polypeptide complex hydrogel (AuNPs/MoS2/Pep hydrogel) with excellent electrochemical performances was created. The anti-fouling electrochemical sweat sensor was fabricated based on the AuNPs/MoS2/Pep hydrogel to address these issues. It was found that the polypeptide hydrogel was designed to form a network structure and carried abundant hydrophilic groups, resulting in a AuNPs/MoS2/Pep hydrogel with superior anti-biofouling properties in sweat for 30 min and even long-term stability in undiluted human sweat. In addition, SEM, TEM, UV, XPS, and infrared spectrogram demonstrated that the binding force of π–π stacking force between MoS2 and naphthalene groups in the designed peptide endowed the polypeptide complex hydrogel with an excellent self-healing property. Furthermore, the polypeptide complex hydrogel preserved wearable device function of continuously monitoring uric acid (UA) and ascorbic acid (AA) in sweat in situ. This novel fabricated sweat sensor with high anti-biofouling ability, excellent self-healing property, and sensitive and selective analytical capability describes a new opportunity for health monitoring in situ.

Molecular and physical probes have been widely employed to investigate physicochemical properties and mechanisms of interfaces due to their ability to provide accurate measurements with temporal and spatial resolution. However, the direct measurement of electroactive species diffusion in ion-selective electrode (ISE) membranes and quantification of the water layer have been challenging due to the high impedance and optical opacity of polymer membranes. In the present work, carbon nanoelectrodes with ultrathin insulating encapsulation and good geometrical structure are reported as physical probes for direct electrochemical measurement of the water layer. The scanning electrochemical microscopy experiment exhibits positive feedback at the interface of the fresh ISE, and negative feedback after conditioning for 3 h. The thickness of the water layer was estimated to be ca. 13 nm. For the first time, we provide direct evidence that, during conditioning, the water molecules diffuse through the chloride ion selective membrane (Cl-ISM) until a water layer establishes at almost 3 h. Furthermore, the diffusion coefficient and concentration of oxygen molecules in the Cl-ISM are also directly electrochemical measured by introducing ferrocene (Fc) as a redox molecule probe. The oxygen concentration in the Cl-ISM decreases during conditioning, suggesting the diffusion of oxygen from ISM to the water layer. The proposed method can be used for the electrochemical measurement of solid contact, providing theoretical guidance and advice for the performance optimization of ISEs.