The 50 most-cited articles in The Analyst, 2020

Optical biosensors have exhibited worthwhile performance in detecting biological systems and promoting significant advances in clinical diagnostics, drug discovery, food process control, and environmental monitoring. Without complexity in their pretreatment and probable influence on the nature of target molecules, these biosensors have additional advantages such as high sensitivity, robustness, reliability, and potential to be integrated on a single chip. In this review, the state of the art optical biosensor technologies, including those based on surface plasmon resonance (SPR), optical waveguides, optical resonators, photonic crystals, and optical fibers, are presented. The principles for each type of biosensor are concisely introduced and particular emphasis has been placed on recent achievements. The strengths and weaknesses of each type of biosensor have been outlined as well. Concluding remarks regarding the perspectives of future developments are discussed.

Liquid chromatography (LC) based techniques in combination with mass spectrometry (MS) detection have had a large impact on the development of new pharmaceuticals in the past decades. Continuous improvements in mass spectrometry and interface technologies, combined with advanced liquid chromatographic techniques for high-throughput qualitative and quantitative analysis, have resulted in a wider scope of applications in the pharmaceutical field. LC-MS tools are increasingly used to analyze pharmaceuticals across a variety of stages in their discovery and development. These stages include drug discovery, product characterization, metabolism studies (in vitro and in vivo) and the identification of impurities and degradation products. The increase in LC-MS applications has been enormous, with retention times and molecular weights (and related fragmentation patterns) emerging as crucial analytical features in the drug development process. The goal of this review is to give an overview of the main developments in LC-MS based techniques for the analysis of small pharmaceutical molecules in…

In recent years, a new group of nanomaterials named nanozymes that exhibit enzyme-mimicking catalytic activity has emerged as a promising alternative to natural enzymes. Nanozymes can address some of the intrinsic limitations of natural enzymes such as high cost, low stability, difficulty in storage, and specific working conditions (i.e., narrow substrate, temperature and pH ranges). Thus, synthesis and applications of hybrid and stimuli-responsive advanced nanozymes could revolutionize the current practice in life sciences and biosensor applications. On the other hand, electrochemical biosensors have long been used as an efficient way for quantitative detection of analytes (biomarkers) of interest. As such, the use of nanozymes in electrochemical biosensors is particularly important to achieve low cost and stable biosensors for prognostics, diagnostics, and therapeutic monitoring of diseases. Herein, we summarize the recent advances in the synthesis and classification of common nanozymes and their application in electrochemical biosensor development. After briefly overviewing the…

Graphene field-effect transistors (GFETs) are emerging as bioanalytical sensors, in which their responsive electrical conductance is used to perform quantitative analyses of biologically-relevant molecules such as DNA, proteins, ions and small molecules. This review provides a detailed evaluation of reported approaches in the design, operation and performance assessment of GFET biosensors. We first dissect key design elements of these devices, along with most common approaches for their fabrication. We compare possible modes of operation of GFETs as sensors, including transfer curves, output curves and time series as well as their integration in real-time or a posteriori protocols. Finally, we review performance metrics reported for the detection and quantification of bioanalytes, and discuss limitations and best practices to optimize the use of GFETs as bioanalytical sensors.

Plasmonic sensors are ideally suited for the design of small, integrated, and portable devices that can be employed in situ for the detection of analytes relevant to environmental sciences, clinical diagnostics, infectious diseases, food, and industrial applications. To successfully deploy plasmonic sensors, scaled-down analytical devices based on surface plasmon resonance (SPR) and localized surface plasmon resonance (LSPR) must integrate optics, plasmonic materials, surface chemistry, fluidics, detectors and data processing in a functional instrument with a small footprint. The field has significantly progressed from the implementation of the various components in specifically designed prism-based instruments to the use of nanomaterials, optical fibers and smartphones to yield increasingly portable devices, which have been shown for a number of applications in the laboratory and deployed on site for environmental, biomedical/clinical, and food applications. A roadmap to deploy plasmonic sensors is provided by reviewing the current successes and by laying out the directions the…

Over the past decade aptamers have emerged as a promising class of bioreceptors for biosensing applications with significant advantages over conventional antibodies. However, experimental studies comparing aptasensors and immunosensors, under equivalent conditions, are limited and the results are inconclusive, in terms of benefits and limitations of each bioreceptor type. In the present work, the performance of aptamer and antibody bioreceptors for the detection of a his-tagged protein, used as a model target, is compared. The bioreceptors are immobilized onto a nanostructured porous silicon (PSi) thin film, used as the optical transducer, and the target protein is detected in a real-time and label-free format by reflective interferometric Fourier transform spectroscopy. For the antibodies, random-oriented immobilization onto the PSi nanostructure results in a poor biosensing performance. Contrary, Fc-oriented immobilization of the antibodies shows a similar biosensing performance to that exhibited by the aptamer-based biosensor, in terms of binding rate, dynamic detection range,…

The ongoing worldwide SARS-CoV-2 epidemic clearly has a tremendous influence on public health. Molecular detection based on oral swabs was used for confirmation of SARS-CoV-2 infection. However, high false negative rates were reported. We describe here the development of a point-of-care (POC) serological assay for the detection of IgG antibody against SARS-CoV-2. The principle of a lateral flow immunoassay strip (LFIAs) consists of fixing SARS-CoV-2 nucleocapsid protein to the surface of the strip and coupling anti-human IgG with colloidal gold nanoparticles (Au NPs). A series of parameters of this method were optimized, including the concentration of coating antigen, BSA blocking concentration and pH value for conjugation. The entire detection process took 15-20 min with a volume of 80 μL of the analyte solution containing 10 μL of serum and 70 μL sample diluent. The performance of the established assay was evaluated using serum samples of the clinically diagnosed cases of Coronavirus Disease 2019 (COVID-19). Our results indicated that the LFIAs for SARS-CoV-2 had satisfactory…

Multiplexed detection of biomarkers, i.e., simultaneous detection of multiple biomarkers in a single assay, is a process of great advantages including enhanced diagnostic precision, improved diagnostic efficiency, reduced diagnostic cost, and alleviated pain of patients. A typical lateral-flow immunoassay (LFIA) is a widely used paper-based immunochromatographic test strip designed to detect a target biomarker through two common formats: sandwich assay and competitive assay. In order to obtain qualitative or quantitative results, a probe with unique optical or magnetic properties is usually employed to characterize the concentration of the target biomarker. The typical LFIA is suitable for point-of-care testing due to its simplicity, portability, cost-effectiveness, and rapid detection of a target biomarker. However, detection of a single biomarker in the typical LFIA is not favorable for high throughput analysis. Therefore, multiplexed detection of biomarkers in LFIAs has been extensively studied in recent years for high throughput analysis. To accomplish multiplexed detection of…

To combat antibiotic resistance, it is extremely important to select the right antibiotic by performing rapid diagnosis of pathogens. Traditional techniques require complicated sample preparation and time-consuming processes which are not suitable for rapid diagnosis. To address this problem, we used surface-enhanced Raman spectroscopy combined with machine learning techniques for rapid identification of methicillin-resistant and methicillin-sensitive Gram-positive Staphylococcus aureus strains and Gram-negative Legionella pneumophila (control group). A total of 10 methicillin-resistant S. aureus (MRSA), 3 methicillin-sensitive S. aureus (MSSA) and 6 L. pneumophila isolates were used. The obtained spectra indicated high reproducibility and repeatability with a high signal to noise ratio. Principal component analysis (PCA), hierarchical cluster analysis (HCA), and various supervised classification algorithms were used to discriminate both S. aureus strains and L. pneumophila. Although there were no noteworthy differences between MRSA and MSSA spectra when viewed with the naked eye,…

Unpredictable and uncontrollable protein adsorption on nanoparticles remains a considerable challenge to achieving effective application of nanotechnologies within biological environments. Nevertheless, engineered nanoparticles offer unprecedented functionality and control in probing and altering biological systems. In this review, we highlight recent advances in harnessing the "protein corona" formed on nanoparticles as a handle to tune functional properties of the protein-nanoparticle complex. Towards this end, we first review nanoparticle properties that influence protein adsorption and design strategies to facilitate selective corona formation, with the corresponding characterization techniques. We next focus on literature detailing corona-mediated functionalities, including stealth to avoid recognition and sequestration while in circulation, targeting of predetermined in vivo locations, and controlled activation once localized to the intended biological compartment. We conclude with a discussion of biocompatibility outcomes for these protein-nanoparticle complexes applied in…

Escherichia coli O157:H7 is a severe foodborne pathogen that causes lots of life-threatening diseases. In the search for a rapid, sensitive, portable and low-cost method to detect this pathogen, we developed a wax-printed paper-based enzyme-linked immunosorbent assay (P-ELISA) based on microfluidic paper-based analytical devices (μPADs), with the whole operation time being less than 3 h and only needing 5 μl samples for detection. The limit of detection (LOD) of E. coli O157:H7 reached 104 CFU ml-1, which is an order of magnitude higher than that of conventional ELISA (C-ELISA). The LOD in artificially contaminated beef samples is 1 CFU per 25 g after enriching the culture for 8 h. This method is superior to the molecular biology method in detection sensitivity and superior to C-ELISA and the national standard method in detection time and cost. Thus, the established P-ELISA method has good sensitivity, specificity and repeatability. It can be suitable for point-of-care testing without expensive and bulky instruments and can also provide a platform for detecting other pathogens,…

Surface-enhanced Raman spectroscopy (SERS) based on machine learning methods has been applied in material analysis, biological detection, food safety, and intelligent analysis. However, machine learning methods generally require extra preprocessing or feature engineering, and handling large-scale data using these methods is challenging. In this study, deep learning networks were used as fully connected networks, convolutional neural networks (CNN), fully convolutional networks (FCN), and principal component analysis networks (PCANet) to determine their abilities to recognise drugs in human urine and measure pirimiphos-methyl in wheat extract in the two input forms of a one-dimensional vector or a two-dimensional matrix. The best recognition result for drugs in urine with an accuracy of 98.05% in the prediction set was obtained using CNN with spectra as input in the matrix form. The optimal quantitation for pirimiphos-methyl was obtained using FCN with spectra in the matrix form, and the analysis was accomplished with a determination coefficient of 0.9997 and a root mean square error…

The combination of electrochemistry and spectroscopy, known as spectroelectrochemistry (SEC), is an already established approach. By combining these two techniques, the relevance of the data obtained is greater than what it would be when using them independently. A number of review papers have been published on this subject, mostly written for experts in the field and focused on recent advances. In this review, written for both the novice in the field and the more experienced reader, the focus is not on the past but on the future. The scope is narrowed down to four techniques the authors claim to have the most potential for the future, namely: infrared spectroelectrochemistry (IR-SEC), Raman spectroelectrochemistry (Raman-SEC), nuclear magnetic resonance spectroelectrochemistry (NMR-SEC) and, perhaps slightly more controversial but certainly promising, electrochemistry mass-spectrometry (EC-MS).

level. The recognition mechanism was based on the catalysis and signal amplification of nanozymes, a new type of nanomaterial possessing specific catalytic activity towards certain chemical reactions such as the oxidation of colorless TMB to blue oxTMB. In this work, we are trying to present readers with a better understanding of this important colorimetric sensing material by illustrating its application in the detection of heavy metal ions using metal nanoparticles, metal oxides, metal sulfides, graphene-based nanomaterials, G-quadruplex, etc. respectively.

Foodborne diseases caused by pathogenic bacteria pose a serious threat to human health. Early and rapid detection of foodborne pathogens is urgently needed. The use of biosensors to identify and detect pathogenic bacteria has attracted ample attention because of their high sensitivity, near real-time quantification without enrichment, on-site detection, simple operation, and so on. As a promising alternative recognition element in biosensors, lectins have been widely studied in bacterial detection because of their high stability and low cost. In this review, we highlight the progress of lectin-based pathogen detection methods, including various electrochemical methods, optical methods and quartz crystal microbalance methods, as well as lectin based microfluidic methods. The interaction mechanism between lectins and bacterial recognition site-sugars is also studied. Finally, the future prospects and challenges in the development of lectin-based biosensors are discussed.

Exosomes are endogenous vesicles of cells, and can be used as important biomarkers for cancers. In this work, we developed a sensitive and reliable SERS sensor for simultaneous detection of multiple cancer-related exosomes. The SERS detection probes were made of bimetallic SERS-active nanotags, gold-silver-silver core-shell-shell nanotrepangs (GSSNTs), which were composed of bumpy surface nanorod (gold nanotrepang, GNT) cores and bilayer silver shells, and decorated with linker DNAs, which were complementary to the aptamer targeting exosomes. Three kinds of SERS detection probes were designed via the adoption of different Raman reporter molecules and linker DNAs. The capture probes were prepared by modifying specific aptamers of the target exosomes on magnetic beads (MBs). In the absence of target exosomes, SERS detection probes were coupled with MBs via specific DNA hybridization for use as aptamer-based SERS sensors. In the presence of target exosomes, the aptamer specifically recognized and captured the exosomes, and GSSNTs were subsequently released into the supernatant.…

Inflammation is a complex biological response of the human body to external or internal stimuli, such as invading pathogens, defective cells, or irritating substances. One important indicator of inflammatory conditions or the progress of various diseases, such as cancer, cardiovascular diseases, neurological diseases, connective tissue diseases, sepsis, or Alzheimer's disease, is the concentration level of inflammatory biomarkers, including immunoglobulins, cytokines, and C-reactive protein (CRP). Since inflammatory biomarkers are highly correlated with each other, it is important to measure them simultaneously. To enable continuous and dynamic inflammatory biomarker detection, we utilized localized surface plasmon resonance (LSPR) to perform label-free molecule sensing. Since the LSPR sensing mechanism requires only a small sensing area with simplified optical setup, it can be easily integrated with a microfluidic device. To simplify reagent operation complexity, we developed an automated microfluidic control system to control reagent guiding and switching in the immunoassay with…

NMR spectroscopy is an essential analytical technique in metabolomics and fluxomics workflows, owing to its high structural elucidation capabilities combined with its intrinsic quantitative nature. However, routine NMR "omic" analytical methods suffer from several drawbacks that may have limited their use as a method of choice, in particular when compared to another widely used technique, mass spectrometry. This review describes, in a critical and perspective discussion, how some of the most recent developments emerging from the NMR community could act as real game changers for metabolomics and fluxomics in the near future. Advanced developments to make NMR metabolomics more resolutive, more sensitive and more accessible are described, as well as new approaches to improve the identification of biomarkers. We hope that this review will convince a broad end-user community of the increasing role of NMR in the "omic" world at the beginning of the 2020s.

Using light as an external stimulus plays a key role not only in modulating activities of nanozymes, but also in constructing efficient biosensing systems. This mini-review highlights recent advances in light-responsive nanozyme-based biosensing systems. First, we introduce the light stimulation for regulating nanozymes' activities. Then, several strategies are presented to construct efficient photo-responsive nanozyme-based detection systems by using metal-based nanozymes, carbon-based nanozymes, and MOF-based nanozymes, respectively. Moreover, the detection mechanisms of current biosensors are discussed. Finally, we discuss the current challenges and future perspectives of this research area.

This work reports the development of a rapid, simple and inexpensive colorimetric paper-based assay for the detection of the severe acute respiratory symptom coronavirus 2 (SARS-CoV-2) humanized antibody. The paper device was prepared with lamination for easy sample handling and coated with the recombinant SARS-CoV-2 nucleocapsid antigen. This assay employed a colorimetric reaction, which is followed by horseradish peroxidase (HRP) conjugated detecting antibody in the presence of the 3,3',5,5'-tetramethylbenzidine (TMB) substrate. The colorimetric readout was evaluated and quantified for specificity and sensitivity. The characterization of this assay includes determining the linear regression curve, the limit of detection (LOD), the repeatability, and testing complex biological samples. We found that the LOD of the assay was 9.00 ng μL-1 (0.112 IU mL-1). The relative standard deviation was approximately 10% for a sample number of n = 3. We believe that our proof-of-concept assay has the potential to be developed for clinical screening of the SARS-CoV-2 humanized antibody as a tool…

Three-dimensional nitrogen-doped graphene (3D-NG) networks, yielded by hydrothermal reaction and freeze-drying treatment, were used as building blocks to construct a metal-free quadruplet electrochemical sensor for simultaneous detection of ascorbic acid (AA), dopamine (DA), uric acid (UA), and acetaminophen (AP). The introduced 3D-NG materials with a 3D porous structure and a nitrogen doping effect were beneficial for the generation of multidimensional electron transfer pathways and the improvement of electrocatalytic activities by modulating their electronic properties, which could contribute to the effective differentiation of the four analytes in their quaternary mixture. Well-resolved oxidation peaks and enhanced response currents of AA, DA, UA, and AP were obtained from the 3D-NG-based electrodes. For the individual determination of one analyte, the linear concentration ranges of AA, DA, UA, and AP were 20-10 000, 1-1000, 0.5-1000, and 0.1-600 μM with detection limits of 3.91, 0.26, 0.12, and 0.02 μM (S/N = 3), respectively. After the synchronous change of the concentrations…

Accelerated development of new therapeutics in an increasingly competitive landscape requires the use of high throughput analytical platforms. In addition, the complexity of novel biotherapeutic formats (e.g. fusion proteins, protein-polymer conjugates, co-formulations, etc.) reinforces the need to improve the selectivity and resolution of conventional one-dimensional (1D) liquid chromatography (LC). Liquid chromatography-mass spectrometry (LC-MS)-based technologies such as native LC-MS for intact mass analysis or peptide mapping (also called bottom-up approach)-based multi-attribute methods (MAM) have already demonstrated their potential to complement the conventional analytical toolbox for monoclonal antibody (mAb) characterization. Two-dimensional liquid-chromatography (2D-LC-MS) methods have emerged in the last ten years as promising approaches to address the increasing analytical challenges faced with novel antibody formats. However, off-line sample preparation procedures are still required for conventional 1D and 2D-LC-MS methods for the in-depth variant characterization at…

for exosomes by detecting three exosomal miRNAs (miR-21, miR-27a and miR-375), respectively. In contrast, there were no obvious correlations between fluorescence intensities and control MCF-10A exosome concentrations. Importantly, by testing multiple exosomal miRNAs using the biosensor in clinical serum samples, breast cancer patients can be effectively differentiated from healthy donors. Consequently, the developed biosensor demonstrates high potential as a routine bioassay for the multiplex quantification of exosomal miRNAs in clinical diagnosis.

The continuing hurdle of developing foodborne pathogen detection techniques is that compromises must be made among simplicity, portability, speed, sensitivity, and quantitation. Herein, we fabricated quantum dot nanobeads (QDNS) by a layer-by-layer assembly of quantum dots on the surface of polymer nanospheres. QDNS exhibited higher fluorescence intensity than the quantum dots at the same particle number. Based on the quantum dot nanobeads as the signal reporter, a quantitative lateral flow immunoassay was demonstrated for Salmonella typhimurium detection with improved sensitivity, specificity and accuracy. A visual detection limit of 5 × 103 CFU mL-1Salmonella typhimurium within 10 min has been proved and demonstrated. Additionally, higher concentrations of non-Salmonella typhimurium bacteria have negligible effects on the detection of Salmonella typhimurium. The results of 50 single blind tests by 10 testers suggested that the assay exhibited 100% accuracy. The results illustrate that the assay provides a balance among simplicity, speed, sensitivity and accuracy, and it can be a…

Establishing standardized methods for a consistent analysis of spectral data remains a largely underexplored aspect in surface-enhanced Raman spectroscopy (SERS), particularly applied to biological and biomedical research. Here we propose an effective machine learning classification of protein species with closely resembled spectral profiles by a mixed data processing based on principal component analysis (PCA) applied to multipeak fitting on SERS spectra. This strategy simultaneously assures a successful discrimination of proteins and a thorough characterization of the chemostructural differences among them, ultimately opening up new routes for SERS evolution toward sensing applications and diagnostics of interest in life sciences.

COVID-19, caused by the infection of SARS-CoV-2, has emerged as a rapidly spreading infection. The disease has now reached the level of a global pandemic and as a result a more rapid and simple detection method is imperative to curb the spread of the virus. We aimed to develop a visual diagnostic platform for SARS-CoV-2 based on colorimetric RT-LAMP with levels of sensitivity and specificity comparable to that of commercial qRT-PCR assays. In this work, the primers were designed to target a conserved region of the RNA-dependent RNA polymerase gene (RdRp). The assay was characterized for its sensitivity and specificity, and validated with clinical specimens collected in Thailand. The developed colorimetric RT-LAMP assay could amplify the target gene and enabled visual interpretation in 60 min at 65 °C. No cross-reactivity with six other common human respiratory viruses (influenza A virus subtypes H1 and H3, influenza B virus, respiratory syncytial virus types A and B, and human metapneumovirus) and five other human coronaviruses (MERS-CoV, HKU-1, OC43, 229E and NL63) was observed.…

Correction for 'Applications of Raman spectroscopy in the development of cell therapies: state of the art and future perspectives' by Shreyas Rangan et al., Analyst, 2020, DOI: 10.1039/c9an01811e.

The monitoring of ammonium ion concentration has gained the attention of researchers from multiple fields since it is a crucial parameter with respect to environmental and biomedical applications. For example, ammonium is considered to be a quality indicator of natural waters as well as a potential biomarker of an enzymatic byproduct in key physiological reactions. Among the classical analytical methods used for the detection of ammonium ions, potentiometric ion-selective electrodes (ISEs) have attracted special attention in the scientific community because of their advantages such as cost-effectiveness, user-friendly features, and miniaturization ability, which facilitate easy portable measurements. Regarding the analytical performance, the key component of ISEs is the selective receptor, labelled as an ionophore in ISE jargon. Indeed, the preference of an ionophore for ammonium amongst other ions (i.e., selectivity) is a factor that primarily dictates the limit of detection of the electrode when performing measurements in real samples. A careful assessment of the literature for…

Despite serving as a gold standard for protein analysis, the classic enzyme-linked immunosorbent assay (ELISA) is currently challenged by the ever-increasing needs of sensitivity and simplicity. Towards the ongoing needs, recent advances in nanomaterials have offered numerous promising tools for enhancing the performance and broadening the applicability of ELISA. In this review, we highlight nanomaterial-enabled strategies that drastically improve the assay performance without significantly altering the classic ELISA format. Particular attention will be focused on the functional roles of nanomaterials as novel readout systems in ELISA, including those serving as substrate-alternatives, enzyme-alternatives, and non-enzymatic signal amplifiers.

Identifying the species and concentrations of antioxidants is really important because antioxidants play important roles in various biological processes and numerous diseases. Compared with an individual sensor detecting a single antioxidant with limited specificity, a sensor array could simultaneously identify various antioxidants, in which 3-5 types of nanomaterials with peroxidase-like activity are absolutely necessary. Herein, as a single-atom nanozyme, Fe-N/C with oxidase-mimicking activity was applied to construct a triple-channel colorimetric sensor array: (1) Fe-N/C catalytically oxidized three substrates 3,3',5,5'-tetramethylbenzidine (TMB), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and o-phenylenediamine (OPD) to produce blue oxidized TMB (oxTMB), green oxidized ABTS (oxABTS) and yellow oxidized OPD (oxOPD), respectively; (2) with oxTMB, oxABTS and oxOPD as three sensing channels, a colorimetric sensor array was constructed for simultaneously discriminating glutathione (GSH), l-cysteine (l-Cys), ascorbic acid (AA), uric acid (UA), and melatonin (MT),…

) to observe the transient response of the sensor when analyte was introduced to the device. The target selectivity of the sensor platform was evaluated and confirmed by challenging the GFET biosensor with various concentrations of lysozyme protein. The results suggest that this device technology has the potential to be used as a general diagnostic platform for measuring clinically relevant biomarkers for point-of-care applications.

Recent advances in the detection of HClO/ClO − in vitro and in vivo using fluorescent, phosphorescent and chemiluminescent probes are reviewed.

Serotonin is a neuromodulator implicated in depression that is often measured in real-time by fast-scan cyclic voltammetry (FSCV). A specialized "Jackson" waveform (JW, 0.2, 1.0 V, -0.1 V, 0.2 V, 1000 V s-1) was developed to reduce serotonin fouling, but the 1.0 V switching potential limits sensitivity and electrodes still foul. The goal of this study was to test the effects of extending the FSCV switching potential to increase serotonin sensitivity and decrease fouling. We compared the Jackson waveform, the dopamine waveform (DA, -0.4 V, 1.3 V, 400 V s-1), and two new waveforms: the extended serotonin waveform (ESW, 0.2, 1.3, -0.1, 0.2, 1000 V s-1) and extended hold serotonin waveform (EHSW, 0.2, 1.3 (hold 1 ms), -0.1, 0.2, 400 V s-1). The EHSW was the most sensitive (LOD = 0.6 nM), and the JW the least sensitive (LOD = 2.4 nM). With the Jackson waveform, electrode fouling was significant with repeated injections of serotonin or exposure to its metabolite, 5-hydroxyindoleacetic acid (5-HIAA). Using the extended waveforms, electrodes fouled 50% less than with the Jackson waveform…

Circulating tumor cells (CTCs) have been widely considered as promising novel biomarkers for molecular research and clinical diagnosis of cancer. However, the sorting of CTCs is very challenging due to the rarity of CTCs in blood and the morphological similarity to blood cells. Although affinity-based CTC sorting methods could capture CTCs using specific biochemical markers, there are limitations such as the loss of cell viability after labeling and a requirement for expensive biochemical marker reagents. Emerging label-free CTC sorting methods rely on the physical properties of cells and can potentially overcome the aforementioned limitations. In this review, we highlight recent advances in label-free CTC sorting methods, with emphasis on device structures and performances. Specifically, we present a detailed discussion on label-free CTC sorting methods, including passive ones that depend on the channel structure or specific fluidic effects and active ones that use external force fields, as well as provide an overview of the principles, advantages, limitations, and applications of…

Nearly 7% of the world's population live with a hemoglobin variant. Hemoglobins S, C, and E are the most common and significant hemoglobin variants worldwide. Sickle cell disease, caused by hemoglobin S, is highly prevalent in sub-Saharan Africa and in tribal populations of Central India. Hemoglobin C is common in West Africa, and hemoglobin E is common in Southeast Asia. Screening for significant hemoglobin disorders is not currently feasible in many low-income countries with the high disease burden. Lack of early diagnosis leads to preventable high morbidity and mortality in children born with hemoglobin variants in low-resource settings. Here, we describe HemeChip, the first miniaturized, paper-based, microchip electrophoresis platform for identifying the most common hemoglobin variants easily and affordably at the point-of-care in low-resource settings. HemeChip test works with a drop of blood. HemeChip system guides the user step-by-step through the test procedure with animated on-screen instructions. Hemoglobin identification and quantification is automatically performed, and…

In most of the sensing systems, specific detection mechanisms are involved during the detection process for a certain analyte irrespective of probes. However, unlike that of various sensing analytes, the detection of the highly toxic and explosive picric acid (PA) analyte was found to involve significant types of distinct sensing mechanisms depending on the nature of probes. Moreover, in the past five years, apart from the plethora of fluorescent probes designed, a number of unique organic small molecules and polymers have been strategically developed at our laboratory for the detection of PA, wherein the involvement of several diverse mechanisms along with a few new mechanisms depending on the electronic and photophysical properties of the probes has been unveiled. This involvement of several distinct mechanisms for the detection of PA motivated us to compile a step-by-step guide for the elucidation of the fluorescence sensing mechanism by taking PA as a model analyte. This "tutorial review" summarizes all the common sensing mechanisms involved for the detection of PA hitherto and…

Neuroinflammation plays a central role in the progression of many neurodegenerative diseases such as Alzheimer's disease, and challenges remain in modeling the complex pathological or physiological processes. Here, we report an acoustofluidic method that can rapidly construct 3D neurospheroids and inflammatory microenvironments for modeling microglia-mediated neuroinflammation in Alzheimer's disease. By incorporating a unique contactless and label-free acoustic assembly, this cell culture platform can assemble dissociated embryonic mouse brain cells into hundreds of uniform 3D neurospheroids with controlled cell numbers, composition (e.g. neurons, astrocytes, and microglia), and environmental components (e.g. amyloid-β aggregates) in hydrogel within minutes. Moreover, this platform can maintain and monitor the interaction among neurons, astrocytes, microglia, and amyloid-β aggregates in real-time for several days to weeks, after the integration of a high-throughput, time-lapse cell imaging approach. We demonstrated that our engineered 3D neurospheroids can represent the amyloid-β…

Machine Learning (ML) has found several applications in spectroscopy, including recognizing minerals and estimating elemental composition. ML algorithms have been widely used on datasets from individual spectroscopy methods such as vibrational Raman scattering, reflective Visible-Near Infrared (VNIR), and Laser-Induced Breakdown Spectroscopy (LIBS). We firstly reviewed and tested several ML approaches to mineral classification from the existing literature, and identified a novel approach for using Deep Learning algorithms for mineral classification from Raman spectra, that outperform previous state-of-the-art methods. We then developed and evaluated a novel method for automatic mineral identification from combining measurements with two complementary spectroscopic methods using Convolutional Neural Networks (CNN) for Raman and VNIR, and cosine similarity for LIBS. Specifically, we evaluated fusing Raman + VNIR, Raman + LIBS or VNIR + LIBS spectra in order to classify minerals. ML methods applied to combined spectral methods presented here are shown to outperform the use of a single…

Nitrate is one of the most common contaminants in food and the environment and mainly arises from intense human activities. Electrochemical sensors have been considered as one of the most promising analytical tools for the rapid detection of nitrate in food and environmental matrices due to their quick response, high sensitivity, ease of operation and miniaturisation, and low sample and power consumption. In this review, we summarise advances in sensors for electrochemical analysis of nitrate over the past decade. We also discuss the application of electrochemical sensing systems for the determination of nitrate in the matrices of fresh water, seawater, food, soil and particulate matter.

This review discusses various bonding strategies for fabricating microfluidic devices, with a special emphasis on the modification of the surface assisted by the use of chemicals to assemble microfluidic devices under mild conditions such as room temperature and atmospheric pressure. The paper includes an overview of the wide ranges of bonding methods used in the fabrication of microfluidic systems, such as anodic bonding, fusion bonding, thermal bonding, solvent bonding, and surface chemical bonding. Among these, surface chemical bonding plays a crucial role between the polydimethylsiloxane (PDMS) and thermoplastics in order to assemble a microfluidic device in a simple and facile manner. The first section reviews relevant research on the sealing for microfluidic devices; in the second section, the materials used for device fabrication are reviewed. The third section discusses the different sealing processes used in the fabrication of microfluidic devices on silicon, glass, thermoplastic, and elastomer. Overall, this review concludes with a discussion on the importance of the…

Chemiluminescence (CL) analysis is a trace analytical method that possesses advantages including high sensitivity, wide linear range, easy operation, and simple instruments. With the development of nanotechnology, many nanomaterial (NM)-enhanced CL systems have been established in recent years and applied for the CL detection of metal ions, anions, small molecules, tumor markers, sequence-specific DNA, and RNA. This review summarizes the research progress of the nanomaterial-enhanced CL systems the past five years. These CL reactions include luminol, peroxyoxalate, lucigenin, ultraweak CL reactions, and so on. The CL mechanisms of the nanomaterial-enhanced CL systems are discussed in the first section. Nanomaterials take part in the CL reactions as the catalyst, CL emitter, energy acceptor, and reductant. Their applications are summarized in the second section. Finally, the challenges and opportunities are discussed.

after 1 min. Moreover, the biosensor was capable of specifically distinguishing S. aureus from other food- and water-borne bacteria such as E. coli and Listeria using the dual mode colorimetric and electrochemical detection. The biosensor was also tested in spiked milk and water samples showing very good recovery percentages. Thus, we believe that this dual mode biosensing platform enables the easy and accurate determination of S. aureus and holds great promise for point-of-care diagnosis.

Fe2+ is a trace metal ion required by the human body, and its abnormal metabolism can cause serious diseases. Herein, we report the development of a highly efficient "ON-OFF" fluorescent probe based on carbon dots (CDs), prepared by a simple one-step hydrothermal method. The CDs exhibited exceptional water dispersibility and stability, superior luminescence performance and low cytotoxicity. The fluorescence could be efficiently quenched by Fe2+ through an electronic transfer process. And under the optimized experimental conditions, this probe shows excellent selectivity and high sensitivity towards Fe2+ with a detection limit of 51 nmol. More interestingly, this probe could realize the visual detection of Fe2+ when Fe3+ and Cu2+ ions were efficiently shielded by tartaric acid in the presence of ascorbic acid. Furthermore, the developed fluorescent probe has been successfully applied for the detection of Fe2+ in tap water and BSA solution as well as for the biosensing of Fe2+ in living cells.

Biothiols play an important role in living cells and are associated with many diseases. Thus, it is necessary to develop a facile, cost-effective, and convenient analytical method for the detection of biothiols. Nanozymes are functional nanomaterials with enzymatic activities. Due to their unique advantages (e.g., low cost, high stability, and multifunctionality), nanozymes have been extensively used to construct sensing systems. Previous studies demonstrated colorimetric assays for biothiol detection because they could competitively inhibit the peroxidase-like activities of nanozymes. However, few studies were able to differentiate biothiols from each other. To address these challenges, herein, we first synthesized Au alloy nanozymes with better peroxidase-like activities than gold nanoparticles (AuNPs). Then, cross-reactive sensor arrays were constructed with three alloy nanozymes. Six typical biothiols (i.e., glutathione, cysteine, dithiothreitol, mercaptoacetic acid, mercaptoethanol, and mercaptosuccinic acid) were successfully detected and discriminated by the as-prepared…

Ferritin, a blood cell protein containing iron, is a crucial biomarker that is used to estimate the risk assessment of iron deficiency anemia. For point-of-care analysis, a reliable, cost-effective, selective, sensitive, and portable tool is extremely necessary. In this study, a label-free electrochemical immunosensor for detecting ferritin using a paper-based analytical device (ePAD) was created. The device pattern was custom designed onto filter paper to successfully fabricate a deliverable immunosensor. Graphene oxide was first modified onto the working electrode using an inkjet printing technique. An activation step of the electrode surface was then performed using standard 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC)/N-hydroxysulfosuccinimide (sulfo-NHS) chemistry. Anti-ferritin antibodies were covalently immobilized onto the amine-reactive ester surface. The amount of ferritin was monitored by observing the electrochemical signal of the selected redox couple by differential pulse voltammetry (DPV). In the presence of ferritin, the sensor showed a considerable decrease…

Functional synapses in the central nervous system depend on a chemical signal exchange process that involves neurotransmitter delivery between neurons and receptor cells in the neuro system. Abnormal neurotransmitter levels and distributions can cause intractable diseases involving descending/ascending modulatory pathways or dysfunctional organs. The detection of abnormal neurotransmitter levels is one of the most promising techniques in the diagnosis of brain diseases. Also, numerous effective methods for the detection of neurotransmitters in vivo have been fabricated. Nowadays, electrochemical, optical, magnetic, and microdialysis methods are among the main techniques used to detect neurotransmitters. Herein, we review current techniques for detecting eight types of neurotransmitters with a focus on in vivo neurotransmitter tracking methods intended for the real-time diagnosis of brain disorders.

Disease diagnostics, food safety monitoring and environmental quality monitoring are the key means to safeguard human health. However, conventional detection devices for health care are costly, bulky and complex, restricting their applications in resource-limited areas of the world. With the rapid development of biosensors and the popularization of smartphones, smartphone-based sensing systems have emerged as novel detection devices that combine the sensitivity of biosensors and diverse functions of smartphones to provide a rapid, low-cost and convenient detection method. In these systems, a smartphone is used as a microscope to observe and count cells, as a camera to record fluorescence images, as an analytical platform to analyze experimental data, and as an effective tool to connect detection devices and online doctors. These systems are widely used for cell analysis, biochemical analysis, immunoassays, and molecular diagnosis, which are applied in the fields of disease diagnostics, food safety monitoring and environmental quality monitoring. Therefore, we discuss four types of…

Nasopharyngeal carcinoma (NPC) is one of the most common malignant tumors in the world, and several studies have associated Epstein-Barr virus (EBV) with NPC occurrence and development. EBV-PCR (polymerase chain reaction), in situ hybridization and immunoassays are the most common methods for NPC identification. However, these approaches have drawbacks, which include tedious procedures and false results. Therefore, a rapid, accurate, and sensitive clinical diagnostic method for the prognosis of EBV-related diseases is needed. In this study, we developed a simple and sensitive approach for EBV detection based on the combination of CRISPR-Cas12a and a lateral flow biosensor (LFB). Cas12a exhibits collateral cleavage propensity of both target DNA and any single-stranded(ss) DNA in the vicinity (herein referred to as a reporter). The LFB test line contained an ssDNA probe complementary to the reporter. In the presence of the target, Cas12a trans-cleaved the ssDNA reporter, which resulted in the inability of cleaved sequences to bind the LFB test line. With a PCR pre-amplification of the…

Food safety and quality regulations inevitably call for sensitive and accurate analytical methods to detect harmful contaminants in food and to ensure safe food for the consumer. Both novel and well-established biorecognition elements, together with different transduction schemes, enable the simple and rapid analysis of various food contaminants. Upconversion nanoparticles (UCNPs) are inorganic nanocrystals that convert near-infrared light into shorter wavelength emission. This unique photophysical feature, along with narrow emission bandwidths and large anti-Stokes shift, render UCNPs excellent optical labels for biosensing because they can be detected without optical background interferences from the sample matrix. In this review, we show how this exciting technique has evolved into biosensing platforms for food quality and safety monitoring and highlight recent applications in the field.

We herein report a simple anthraquinone-imidazole-based compound, 2-(furan-2-yl)-1H-anthra[1,2-d]imidazole-6,11-dione (FAI), as an effective colorimetric and fluorometric Ag+ sensor. In 1,4-dioxane/water (3 : 7, v/v, pH 7.4), FAI could selectively recognize and detect Ag+ ions via significant fluorescence quenching at 606 nm, accompanied by a rapid color change from yellow to orange. FAI exhibited high sensitivity to Ag+ in a linear dynamic range from 0.8-32 μM, with the detection limit of 66 nM. Job's plot analysis and HRMS studies supported the formation of the FAI-Ag+ complex in a ratio of 1 : 1. The in situ generated FAI-Ag+ complex was further utilized as a reversible sensor to detect biothiols (Cys, Hcy, GSH) through a decomplexation reaction, with the detection limit of 89 nM, 174 nM and 208 nM for Cys, Hcy and GSH, respectively. In addition, FAI with low cytotoxicity was efficient in monitoring Ag+ ions and biothiols in living human hepatoma cells SMMC-7721 as demonstrated by fluorescence microscopy imaging.