The 100 most-cited articles in Analytical Chemistry, 2022

This work presented a point-of-care (POC) photoelectrochemical (PEC) biosensing for the detection of human papillomavirus-16 (HPV-16) on a portable electrochemical detection system by using CRISPR-Cas12a trans-cleaving the G-quadruplex for the biorecognition/amplification and a hollow In 2 O 3 –In 2 S 3 -modified screen-printed electrode (In 2 O 3 –In 2 S 3 /SPE) as the photoactive material. G-quadruplexes were capable of biocatalytic precipitation (H 2 O 2 -mediated 4-chloro-1-naphthol oxidation) on the In 2 O 3 –In 2 S 3 /SPE surface, resulting in a weakened photocurrent, but suffered from trans-cleavage when the CRISPR-Cas12a system specifically recognized the analyte. The photocurrent results could be directly observed with the card-sized electrochemical device via a smartphone, which displayed a high-value photocurrent for these positive samples, while a low-value photocurrent for the target-free samples. Such a system exhibited satisfying photocurrent responses toward HPV-16 within a wide working range from 5.0 to 5000 pM and allowed for detection of HPV-16 at a concentration…

Large-scale, rapid, and inexpensive serological diagnoses of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) are of great interest in reducing virus transmission at the population level; however, their development is greatly plagued by the lack of available point-of-care methods, leading to low detection efficiency. Herein, an ultrasensitive smartphone-based electrochemical immunoassay is reported for rapid (less than 5 min), low-cost, easy-to-implement detection of the SARS-CoV-2 nucleocapsid protein (SARS-CoV-2 N protein). Specifically, the electrochemical immunoassay was fabricated on a screen-printed carbon electrode coated with electrodeposited gold nanoparticles, followed by incubation of anti-N antibody (Ab) and bovine serum albumin as the working electrode. Accompanied by the antigen–antibody reaction between the SARS-CoV-2 N protein and the Ab, the electron transfer between the electroactive species [Fe(CN) 6 ] 3– / 4– and the electrode surface is disturbed, resulting in reduced square-wave voltammetry currents at 0.075 V versus the Ag/AgCl reference electrode.…

Common homogeneous electrochemical (HEC) sensors usually suffer from the drawbacks of high background signal, low signal-to-noise ratio, and even false positive results due to the preaddition of electroactive substances. Thus, it is necessary to develop novel HEC sensors based on in situ generation of electroactive substances to overcome these shortcomings, which, however, is underexplored. In this work, two-dimensional (2D) nanozymes, i.e., cobalt-doped 2D Ti 3 C 2 MXene nanosheets (CMNSs), with excellent peroxidase-like properties were utilized to develop HEC sensors based on the in situ generation of electroactive substances for organophosphate pesticides (OPs) detection. The 2D CMNSs were synthesized via a template-directed wet chemical approach and displayed outstanding features of hydrophilia and water dispersibility, which could catalyze the oxidation of o -phenylenediamine (OPD) to generate significantly increased reduction current. Interestingly, the 2D CMNSs with peroxidase-like properties exhibited a unique response to thiol compounds and were thus employed as highly…

Exploiting innovative sensing mechanisms and their rational implementation for selective and sensitive detection has recently become one of the mainstream research directions of photoelectrochemical (PEC) bioanalysis. In contrast to existing conventional strategies, this study presents a new liposome-mediated method via in situ combining ZnInS nanosheets (ZIS NSs) with SnS 2 to form a ZIS NSs/SnS 2 type-I heterojunction on fluorine-doped tin oxide (FTO) electrodes for highly sensitive PEC immunoassays. Specifically, alkaline phosphatase (ALP)-encapsulated liposomes were confined within 96-well plates by sandwich immunorecognition and subsequently subjected to lysis treatment. Enzymatically produced H 2 S by the released ALP was then directed to react with Sn(IV) to engender the ZIS NSs/SnS 2 type-I heterojunction on the FTO/ZIS NSs-Sn(IV) electrode, resulting in a change in the photogenerated electron–hole transfer path of the photoelectrode and reduction in current signaling. Exemplified by heart-type fatty acid binding protein (h-FABP) as a target, the constructed PEC sensor…

CRISPR/Cas12, a highly efficient and specific nucleic acid recognition system, has been broadly employed to detect amplified DNA products. However, most reported methods adopt a two-step detection mode that needs a liquid transfer step, thus complicating the detection procedure and posing a risk of aerosol contamination. A one-pot detection method can obviate these problems, but it suffers from poor detection efficiency due to the loss of amplification templates elicited by CRISPR/Cas12 cleavage. In this study, we discovered that a glycerol additive dramatically promoted the detection efficiency of the one-pot recombinase polymerase amplification (RPA)-CRISPR/Cas12a method. Compared with the glycerol-free version, its sensitivity was nearly 100-fold higher and was close to that of the canonical two-step method. Further investigation displayed that the enhanced detection efficiency was attributed to the phase separation of the RPA and CRISPR/Cas12a system during the initial phase of the RPA reaction caused by the glycerol viscosity. This highly efficient one-pot method has been…

Distinguished by the coupled catalysis-facilitated high turnover and admirable specificity, enzyme cascades have sparked tremendous attention in bioanalysis. However, three-enzyme cascade-based versatile platforms have rarely been explored without resorting to tedious immobilization procedures. Herein, we have demonstrated that formamide-converted transition metal–nitrogen–carbon (f-MNC, M = Fe, Cu, Mn, Co, Zn) with a high loading of atomically dispersed active sites possesses intrinsic peroxidase-mimetic activity following the activity order of f-FeNC > f-CuNC > f-MnNC > f-CoNC > f-ZnNC. Ulteriorly, benefitting from the greatest catalytic performance and explicit catalytic mechanism of f-FeNC, versatile enzyme cascade-based colorimetric bioassays for ultrasensitive detection of diabetes-related glucose and α-glucosidase (α-Glu) have been unprecedentedly devised using f-FeNC-triggered chromogenic reaction of 3,3′,5,5′-tetramethylbenzidine as an amplifier. Notably, several types of α-Glu substrates can be effectively utilized in this three-enzyme cascade-based α-Glu assay, and it can…

Point-of-care testing (POCT) technology has made major breakthroughs in community medicine and physician office situations, in tandem with the more ubiquitous and intensive usage of highly integrated quick detection equipment for illness diagnosis, personal care, and mobile healthcare. Although the photoelectrochemical (PEC)-based POCT platform offers the benefits of cheap cost and good user engagement, its commercialization is still limited by the photodetection components’ downsizing and mobility, among other factors. In this work, a novel highly integrated PEC biosensor aided by piezophototronics to enhance the efficiency of PEC testing was reported for flexible detection of cancer-associated antigens in biological fluids (prostate-specific antigen, PSA, used as an example). Multiple signal enhancement strategies, including a magnetic bead-linked enzyme-linked immune system catalyzing the production of ascorbic acid from the substrate and a piezoelectric-assisted enhancement strategy, were used for sensitive detection of the analyte to be tested in human body fluids. Unlike the…

The ability to continuously monitor the concentration of specific molecules in the body is a long-sought goal of biomedical research. For this purpose, interstitial fluid (ISF) was proposed as the ideal target biofluid because its composition can rapidly equilibrate with that of systemic blood, allowing the assessment of molecular concentrations that reflect full-body physiology. In the past, continuous monitoring in ISF was enabled by microneedle sensor arrays. Yet, benchmark microneedle sensors can only detect molecules that undergo redox reactions, which limits the ability to sense metabolites, biomarkers, and therapeutics that are not redox-active. To overcome this barrier, here, we expand the scope of these devices by demonstrating the first use of microneedle-supported electrochemical, aptamer-based (E-AB) sensors. This platform achieves molecular recognition based on affinity interactions, vastly expanding the scope of molecules that can be sensed. We report the fabrication of microneedle E-AB sensor arrays and a method to regenerate them for multiple uses. In addition, we…

Recently emerged Z-scheme heterostructure-based immunoassays have presented new opportunities for photoelectrochemical (PEC) biosensing development. Here, we described a tunable signal-on PEC biosensor for the detection of cardiac troponin I (cTnI), which exploited a competitive absorption effect between Cu(II) ions and a Zr metal–organic framework (Zr-MOF) constructed on TiO 2 nanorods (Cu 2+ @Zr-MOF@TiO 2 NRs). Water-stable Zr-MOF was coated onto TiO 2 NRs on fluorine-doped tin oxide to form a Z-scheme heterostructure substrate (Zr-MOF@TiO 2 NRs), which exhibited a high photoelectric response. Cu 2+ @Zr-MOF@TiO 2 NRs, constructed by loading Cu(II) ions onto the architecture of Zr-MOF by electrostatic interaction, demonstrated a low background signal. After sandwich immunorecognition within a 96-well plate, H 2 S, generated by confined alkaline phosphatase on zeolitic imidazolate framework-8, was directed to react with Cu(II) ions to form CuS. This resulted in an in situ change in the photoelectrode and an enhanced photoelectric signal. The developed PEC biosensing platform…

As a golden partner of recombinase polymerase amplification (RPA), CRISPR/Cas12a has been proven to solve the false-positive problem caused by nonspecific amplification perfectly; meanwhile, its trans -cleave activity has further enhanced the sensitivity. However, the solution transfer operation after tube cap opening greatly increases the risk of aerosol contamination of amplicon, which is inconsistent with point-of-care (POC) diagnostics requirements. This study proposes a photoactivated CRISPR/Cas12a strategy to achieve one-pot high-sensitivity nucleic acid detection. Using photocleavable complementary ssDNA to block crRNA, RPA amplification can smoothly pass through the exponential interval without being affected by activated Cas12a in the critical early stage. After enough amplicons were produced, the Cas12a test was activated by short bursts of ultraviolet radiation at 365 nm. This one-pot method achieved a sensitivity of 2.5 copies within 40 min. This simple and sensitive one-pot method can effectively avoid amplicon contamination and lower the threshold for molecular…

Biomimic nanozymes coassembled by peptides or proteins and small active molecules provide an effective strategy to design attractive nanozymes. Although some promising nanozymes have been reported, rational regulation for higher catalytic activity of biomimic nanozymes remains challenging. Hence, we proposed a novel biomimic nanozyme by encapsulating the coassembly of hemin/bovine serum albumin (BSA) in zeolite imidazolate frameworks (ZIF-8) to achieve controllable tailoring of peroxidase-like activity via the confinement effect. The assembly of Hemin@BSA was inspired by the structure of horseradish peroxidase (HRP), in which hemin served as the active cofactor surrounded by BSA as a blocking pocket to construct a favorable hydrophobic space for substrate enrichment. Benefiting from the confinement effect, ZIF-8 with a porous intracavity was identified as the ideal outer layer for Hemin@BSA to accelerate substrate transport and achieve internal circulation of peroxidase-like catalysis, significantly enhancing its peroxidase-like activity. Especially, the precise encapsulation of…

This work reports a contactless photoelectrochemical biosensor based on an ultraviolet-assisted gas sensor (UV–AGS) with a homemade three-dimensional (3D)-SnS 2 nanosheet-functionalized interdigitated electrode. After rigorous examination, it was found that the gas responsiveness accelerated and the sensitivity increased using the UV irradiation strategy. The effects of the interlayer structure and the Schottky heterojunction on the gas-sensitive response of O 2 and NH 3 under UV irradiation were further investigated theoretically by 3D electrostatic field simulations and first-principles density functional theory to reveal the mechanism. Finally, a UV–AGS device was developed to quantify the blood ammonia bioassay in a small-volume whole blood sample by alkalizing blood to release gas-phase ammonia with a linear range of 25–5000 μM with a limit of detection (LOD) of 29.5 μM. The device also enables a rapid immunoassay of human cardiac troponin I (cTnI) with a linear range of 0.4–25.6 ng/mL and an LOD of 0.37 ng/mL using a urease-labeled antibody as the immune recognition molecule.…

Meat’s freshness is closely related to food safety and human health and has received increasing attention nowadays. To on-site visually screen meat freshness in a fast and non-destructive manner, we rationally constructed a series of fluorescent probes ( JDCN, JDNS, and JDPY ) with distinct electron-withdrawing substitution groups based on julolidine-fused coumarin. These probes underwent an aza-Michael addition followed by an elimination reaction with cadaverine to generate a colorimetric and ratiometric fluorescence response, and their sensing performance was rationally enhanced by improving the electron-withdrawing strength of substitution groups. Particularly, JDCN with a dicyanovinyl group as the reaction site exhibited outstanding sensing performance including rapid response (∼60 s), high selectivity, and low detection limit (14 nM). Furthermore, JDCN was fabricated into test kits to detect cadaverine vapor with a high-contrast fluorescence change from red to green. Based on two-color visualization of cadaverine vapor, on-site non-contact and non-destructive monitoring of meat…

Interest in CRISPR-Cas12 and CRISPR-Cas13 detection continues to increase as these detection schemes enable the specific recognition of nucleic acids. The fundamental sensitivity limits of these schemes (and their applicability in amplification-free assays) are governed by kinetic rates. However, these kinetic rates remain poorly understood, and their reporting has been inconsistent. We quantify kinetic parameters for several enzymes (LbCas12a, AsCas12a, AapCas12b, LwaCas13a, and LbuCas13a) and their corresponding limits of detection (LoD). Collectively, we present quantification of enzyme kinetics for 14 guide RNAs (gRNAs) and nucleic acid targets for a total of 50 sets of kinetic rate parameters and 25 LoDs. We validate the self-consistency of our measurements by comparing trends and limiting behaviors with a Michaelis–Menten trans -cleavage reaction kinetics model. For our assay conditions, activated Cas12 and Cas13 enzymes exhibit trans -cleavage catalytic efficiencies between order 10 5 and 10 6 M –1 s –1 . For assays that use fluorescent reporter molecules (ssDNA and ssRNA)…

Immunoglobulin detection is essential for diagnosing progression of SARS-CoV-2 infection, for which SARS-CoV-2 IgG is one of the most important indexes. In this paper, Ag nanoparticles with ultrathin Au shells (∼2 nm) embedded with 4-mercaptobenzoic acid (MBA) (Ag MBA @Au) were manufactured via a ligand-assisted epitaxial growth method and integrated into lateral flow immunoassay (LFIA) for colorimetric and SERS dual-mode detection of SARS-CoV-2 IgG. Ag MBA @Au possessed not only the surface chemistry advantages of Au but also the superior optical characteristics of Ag. Moreover, the nanogap between the Ag core and the Au shell also greatly enhanced the Raman signal. After being modified with anti-human antibodies, Ag MBA @Au recognized and combined with SARS-CoV-2 IgG, which was captured by the SARS-CoV-2 spike protein on the T line. Qualitative analysis was achieved by visually observing the color of the T line, and quantitative analysis was conducted by measuring the SERS signal with a sensitivity four orders of magnitude higher (detection limit: 0.22 pg/mL). The intra-assay and…

Hydrogen sulfide (H 2 S) is an important endogenous gas signal molecule in living system, which participates in a variety of physiological processes. Very recent evidence has accumulated to show that endogenous H 2 S is closely associated with various cancers and can be regarded as a biomarker of cancer. Herein, we have constructed a new near-infrared fluorescent probe (DCP-H 2 S) based on isophorone-xanthene dye for sensing hydrogen sulfide (H 2 S). The probe shows remarkable NIR turn-on signal at 770 nm with a large Stokes shift of 200 nm, together with high sensitivity (15-fold) and rapid detection ability for H 2 S (4 min). The probe also possesses excellent selectivity for H 2 S over various other analytes including biothiols containing sulfhydryl (−SH). Moreover, DCP-H 2 S has been successfully applied to visualize endogenous and exogenous H 2 S in living cells (293T, Caco-2 and CT-26 cells). In particular, the excellent ability of DCP-H 2 S to distinguish normal mice and tumor mice is shown, and it is expected to be a powerful tool for detection of H 2 S in cancer diagnosis.

Functional photothermal nanomaterials have gained widespread attention in the field of precise cancer therapy and early disease diagnosis due to their unique photothermal conversion properties. However, the relatively narrow temperature response range and the outputable accuracy of commercial thermometers limit the accurate detection of biomarkers. Herein, we designed a liposome-embedded Cu 2– x Ag x S amplification-based photothermal sensor for the accurate determination of cardiac troponin I (cTnI) in health monitoring and point-of-care testing (POCT). The combinable 3D-printing detecting device monitored and visualized target signal changes in the testing system under the excitation of near-infrared (NIR) light, which was recorded and evaluated for possible pathogenicity by a smartphone. Notably, we predicted the potentially efficient thermal conversion efficiency of Cu 2– x Ag x S from the structure and charge density distribution, calculated by the first-principles and density functional theory (DFT), which provided a theoretical basis for the construction of novel photothermal…

Great enthusiasm in single-atom catalysts for various catalytic reactions continues to heat up. However, the poor activity of the existing single/dual-metal-atom catalysts does not meet the actual requirement. In this scenario, the precise design of triple-metal-atom catalysts is vital but still challenging. Here, a triple-atom site catalyst of FeCoZn catalyst coordinated with S and N, which is doped in the carbon matrix (named FeCoZn-TAC/SNC), is designed. The FeCoZn catalyst can mimic the activity of oxidase by activating O 2 into • O 2 – radicals by virtue of its atomically dispersed metal active sites. Employing this characteristic, triple-atom catalysts can become a great driving force for the development of novel biosensors featuring adequate sensitivity. First, the property of FeCoZn catalyst as an oxidase-like nanozyme was explored. The obtained FeCoZn-TAC/SNC shows remarkably enhanced catalytic performance than that of FeCoZn-TAC/NC and single/dual-atom site catalysts (FeZn, CoZn, FeCo-DAC/NC and Fe, Zn, Co-SAC/NC) because of trimetallic sites, demonstrating the synergistic…

Two kinds of two-dimensional (2D) covalent-organic frameworks (COF) were used to construct a sandwich-type electrochemical immunosensor for a proof-of-concept study. Vinyl-functionalized COFTab-Dva could be linked with Ab1 by the thiol-ene "click" reaction. Electroactive COFTFPB-Thi was modified with gold nanoparticles (AuNPs) to ensure the successful connection with Ab2 through Au-S bond. Meanwhile, electroactive COFTFPB-Thi was used to as signal probe to realize both the detection of carcinoembryonic antigen (CEA) and the amplification of detection signal. In detection process of the sandwich-type electrochemical immunosensor, glassy carbon electrode (GCE) was modified with 2D COFTab-Dva first then connected with Ab1 by the thiol-ene "click" reaction, next quantitative CEA was captured, followed by specificially capturing signal probe of Ab2/AuNPs/COFTFPB-Thi where AuNPs acted as nanocarriers of Ab2 and COFTFPB-Thi served as the signal producers. As the amount of CEA was increased, the amount of signal probe captured to the electrode was also increased, and the peak signal…

High Resolution Image Download MS PowerPoint Slide The inner filter effect (IFE) hinders fluorescence measurements, limiting linear dependence of fluorescence signals to low sample concentrations. Modern microplate readers allow movement of the optical element in the vertical axis, changing the relative position of the focus and thus the sample geometry. The proposed Z -position IFE correction method requires only two fluorescence measurements at different known vertical axis positions ( z -positions) of the optical element for the same sample. Samples of quinine sulfate, both pure and in mixtures with potassium dichromate, showed a linear dependence of corrected fluorescence on fluorophore concentration ( R 2 > 0.999), up to A ex ≈ 2 and A em ≈ 0.5. The correction extended linear fluorescence response over ≈98% of the concentration range with ≈1% deviation of the calibration slope, effectively eliminating the need for sample dilution or separate absorbance measurements to account for IFE. The companion numerical IFE correction method further eliminates the need for any geometric…

Photo-assisted biofuel cell-based self-powered biosensors (PBFC-SPBs) possess the advantages of no need for external power supply, ease of sensing design, and simple instruments. In this work, a robust anti-interference PBFC-SPB for microRNA detection was constructed based on the Pt–S bond and the inorganic–organic hybridization strategy. The organic semiconductor [6,6]-phenyl-C61-butyric acid methylester@anthraquinone (PCBM@anthraquinone) served as an efficient light-harvesting material, and gold nanoparticle@Pt (AuNP@Pt) nanomaterials were immobilized on the surface via electrostatic adsorption for the binding of DNA. Notably, compared to Au–S bonds for DNA immobilization, the Pt–S bond exhibited better anti-interference ability. Ingeniously, cadmium sulfide quantum dots (CdS QDs) were close to the PCBM@anthraquinone substrate electrode to form sensitization structures, which was beneficial to enhance the photocurrent signal. Combining with the laccase-mimicking activity Cu2+/carbon nanotubes (Cu2+/CNTs) cathode, the PBFC-SPB for microRNA detection was achieved. Once the target…

Complete, reproducible extraction of protein material is essential for comprehensive and unbiased proteome analyses. A current gold standard is single-pot, solid-phase-enhanced sample preparation (SP3), in which organic solvent and magnetic beads are used to denature and capture proteins, with subsequently washes allowing contaminant removal. However, SP3 is dependent on effective protein immobilisation onto beads, risks losses during wash steps, and experiences a drop-off in protein recovery at higher protein inputs. Magnetic beads may also contaminate samples and instruments, and become costly for larger scale protein preparations. Here, we propose solvent precipitation SP3 (SP4) as an alternative to SP3, omitting magnetic beads and employing brief centrifugation—either with or without low-cost inert glass beads—as the means of aggregated protein capture. SP4 recovered equivalent or greater protein yields for 1–5000 µg preparations and improved reproducibility (median protein R 2 0.99 (SP4) vs . 0.97 (SP3)). Deep proteome profiling (n = 9,076) also demonstrated improved recovery…

Herein, we developed a flexible, low-cost thermosensitive fiber paper for the visual display in photothermal biosensing systems for early acute myocardial infarction. The thermal signal visualization device was encapsulated with rewritable thermal fibers, which exhibited excellent stability and reversibility. The mechanism of color change in thermal paper was based on a temperature-driven reversible transformation of the structure of the dye molecule (crystalline violet lactone, CVL). It exhibits a gradation from blue to colorless at higher temperatures and gradually returns to blue when the temperature drops. Immobilization and cascade enzymatic reactions of target molecules occurred in an integrated 3D-printed detection device, a photothermal conversion process occurred under near-infrared light excitation, and the colorimetric change values of the encapsulated thermal paper were recorded and evaluated for possible pathogenicity using a smartphone. It was worth noting that the effect of the thermogenic ring-opening behavior of CVL on the macroscopic phenomenon of color change was…

The clustered regularly interspaced short palindromic repeats (CRISPR)-based nucleic acid detection can be combined with recombinase-aided amplification (RAA) to enable rapid, accurate, and early detection of SARS-CoV-2. Current CRISPR-based approaches to detecting viral nucleic acid typically require immense manual operations to transfer RPA amplicons for CRISPR detection or suffer from compromised sensitivity by mixing the competing RPA amplification and CRISPR detection. Here, we develop dual-CRISPR/Cas12a-assisted RT-RAA assay and a ″sample-to-answer″ centrifugal microfluidic platform that can automatically detect 1 copy/μL of the SARS-CoV-2 within 30 min. This chip separates the amplification (RAA) from detection (CRISPR), such that sensitivity is maximized and the time consumption is decreased by a factor of 3. For the 26 positive and 8 negative clinical SARS-CoV-2 samples, this automated centrifugal microfluidics achieved 100% accuracy compared to the gold-standard RT-PCR technique. This point-of-care test, with the advantages of being one-step, automated, rapid, and…

Herein, we developed a flexible, low-cost non-enzymatic sweat sensing chip for in situ acquisition of bioinformation in sweat of individuals under exercise conditions to advance personal health monitoring and medication management for patients with Parkinson’s disease. This low-cost, flexible, wearable sweat sensor consists of a printed screen electrode modified with g-C 3 N 4 material and an external MSME element. The doping strategy and surface activation strategy of the g-C 3 N 4 -based exhibited efficient glucose oxidase-like activity and electrochemical activity when testing l -dopa and glucose in sweat. The optimized signal was transmitted to a smartphone for processing 12 individuals with simulated dosing, enabling continuous monitoring of l -dopa metabolism in sweat and management of dosing. The generalization ability and robustness of models constructed by methods such as multiple linear regression, artificial neural networks, and convolutional neural networks were compared cross-sectionally. Deep learning models based on artificial neural networks help develop a…

Visual one-step simultaneous detection of low-abundance methylation is a crucial challenge in early cancer diagnosis in a simple manner. Through the design of a closed split bipolar electrochemistry system (BE), detection of promoter methylation of tumor suppressor genes in papillary thyroid cancer, RASSF1A and SLC5A8, was achieved using electrochemiluminescence. For this purpose, electrochemiluminescence of luminol loaded into the Fe 3 O 4 @UiO-66 and gold nanorod-functionalized graphite-like carbon nitride nanosheet (AuNRs@C 3 N 4 NS), separately, on the anodic and cathodic pole bipolar electrodes (BPEs) in two different chambers of a bipolar cell were recorded on a smartphone camera. To provide the same electric potential (Δ E elec ) through the BPEs to conduct simultaneous light emission, as well as to achieve higher sensitivity, anodic and cathodic poles BPEs were separately connected to ruthenium nanoparticles electrodeposited on nitrogen-doped graphene-coated Cu foam (fCu/N-GN/RuNPs) to provide a hydrogen evolution reaction (HER) and polycatechol-modified reduced graphene…

Sweat pH is a critical indicator for evaluating human health. With the extensive attention on the wearable and flexible biosensing devices, the technology for the monitoring of human sweat can be realized. In this study, a sensitive, miniaturized, and flexible electrochemical sweat pH sensor was developed for the continuous and real-time monitoring of the hydrogen-ion concentration in human sweat. A flexible electrode was fabricated on the poly(ethylene terephthalate) (PET) substrate by a simple and low-cost screen-printing technology, which was based on the integration of fluoroalkyl silane-functionalized Ti 3 C 2 T x (F–Ti 3 C 2 T x ) and the polyaniline (PANI) membrane technology instead of the traditional ion-sensitive membrane. The surface functionalization strategy for Ti 3 C 2 T x with perfluorodecyltrichlorosilane can provide environmental stability. Functionalized Ti 3 C 2 T x (F–Ti 3 C 2 T x ) was doped with PANI to obtain improved responsiveness, sensitivity, and reversibility. The constructed microsize, portable, and wearable F–Ti 3 C 2 T x /PANI pH sensor aimed to…

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has lasted for almost 2 years. Stemming its spread has posed severe challenges for clinical virus detection. A long turnaround time, complicated operation, and low accuracy have become bottlenecks in developing detection techniques. Adopting a direct antigen detection strategy, we developed a fast-responding and quantitative capacitive aptasensor for ultratrace nucleocapsid protein detection based on a low-cost microelectrode array (MEA) chip. Employing the solid–liquid interface capacitance with a sensitivity of picofarad level, the tiny change on the MEA surface can be definitively detected. As a result, the limit of detection reaches an ultralow level of femtogram per milliliter in different matrices. Integrated with efficient microfluidic enrichment, the response time of this sensor from the sample to the result is shortened to 15 s, completely meeting the real-time detection demand. Moreover, the wide linear range of the sensor is from 10 –5 to 10 –2 ng/mL, and a high selectivity of 6369:1 is…

Tetracyclines are a group of important antibiotics with a common four-ring scaffold. While most tetracyclines are currently used only in animals, their leaching into the environment and residues in food have caused health concerns. Aptamers are an attractive way to detect tetracyclines, and all previously reported aptamers for tetracyclines were obtained by immobilizing target molecules. In this work, we selected a few DNA aptamers by immobilizing the DNA library using oxytetracycline as the target. We obtained new aptamers with no overlapping sequences compared to the previously reported ones, and a representative sequence named OTC5 had a dissociation constant of 147 nM measured by isothermal titration calorimetry. Similar binding affinities were also observed with tetracycline and doxycycline. Because tetracyclines are fluorescent and their fluorescence intensity was enhanced by binding to the aptamers, a label-free and dye-free fluorescent biosensor was developed with a detection limit of 25 nM oxytetracycline. The sensor was able to detect targets in milk after extraction.…

In this work, a novel ratio electrochemical biosensing platform based on catalytic hairpin assembly target recovery to trigger dual-signal output was developed for ultrasensitive detection of microRNA (miRNA). To achieve the ratiometric dual-signal strategy, methylene blue (MB), an electrochemical indicator, was ingeniously loaded into the pores of graphene aerogel (GA) and metal–organic framework (MOF) composites with high porosity and large specific surface area, and another electrochemical indicator Fe-MOFs with distinct separation of redox potential was selected as a signal probe. Concretely, with the presence of the target miRNA, the CHA process was initiated and the signal probe was introduced to the electrode surface, producing abundant double-stranded H1-H2@Fe-MOFs-NH 2 . Then, the measurement and analysis of the prepared ratiometric electrochemical biosensor by differential pulse voltammetry (DPV) showed that the introduction of the target miRNA led to an increase in the oxidation peak signal of Fe-MOFs (+0.8 V) and a decrease in the oxidation peak signal of MB (−0.23 V).…

The conventional cathodic electrochemiluminescence (ECL) always requires a more negative potential to trigger strong emission, which inevitably damages the bioactivity of targets and decreases the sensitivity and specificity. In this work, iron single-atom catalysts (Fe–N–C SACs) were employed as an efficient co-reaction accelerator for the first time to achieve the impressively cathodic emission of a luminol–H 2 O 2 ECL system at an ultralow potential. Benefiting from the distinct electronic structure, Fe–N–C SACs exhibit remarkable properties for the activation of H 2 O 2 to produce massive reactive oxygen species (ROS) under a negative scanning potential from 0 to −0.2 V. The ROS can oxidize the luminol anions into luminol anion radicals, avoiding the tedious electrochemical oxidation process of luminol. Then, the in situ -formed luminol anion radicals will directly react with ROS for the strong ECL emission. As a proof of concept, sensitive detection of the carcinoembryonic antigen was realized by glucose oxidase-mediated ECL immunoassay, shedding light on the superiority of…

Hypochlorite (ClO – ) and peroxynitrite (ONOO – ) are two crucial highly reactive oxygen/nitrogen species, which interplay with each other, and are implicated in numerous pathophysiological processes. The simultaneous detection of ClO – and ONOO – is immensely significant in evaluating the occurrence and progress of related diseases. Herein, a dual-responsive ratiometric fluorescent probe PTZ-H for the separate and simultaneous detection of ClO – and ONOO – was designed and synthesized. In this probe, the phenothiazine-based coumarin moiety was chosen as the ClO – responsive fluorescent fragment, and the precursor of 2-(benzo[ d ]thiazol-2-yl)aniline was employed as the sensor for ONOO – . The PTZ-H emitted red fluorescence (640 nm) can switch to green (520 nm) and turn on blue fluorescence (450 nm) in response to ClO – and ONOO –, respectively. This allowed the specific recognition and ratiometric quantification of ClO – and ONOO – with the detection limits of 17 and 21 nM, respectively. Notably, confocal laser scanning microscopy revealed that the PTZ-H probe could…

The prominent pathological feature of fatty liver disease lesions is excessive fat accumulation in lipid droplets in hepatocytes. Thus, developing fluorescent lipid droplet-specific probes with high permeability and a high imaging contrast provides a robust tool for diagnosing fatty liver diseases. Herein, we rationally developed a novel donor-acceptor lipophilic fluorescent probe ANI with high photostability for wash-free visualization of lipid droplets and fatty liver disease characteristics. ANI showed a typical twisted intramolecular charge transfer effect with very faint fluorescence in high-polar solvents, but dramatically boosted emissions in low-polar environments. The solvatochromic probe can selectively light up lipid droplets with a high contrast in a wash-free manner. Further use of ANI to reveal the excessive accumulation of lipid droplets with a significantly large size in the liver tissues from the fatty liver disease model mice was successfully demonstrated. The remarkable imaging performances rendered ANI an alternative tool for accurately evaluating fatty liver…

High Resolution Image Download MS PowerPoint Slide Conventional nanomaterials in electrochemical nonenzymatic sensing face huge challenge due to their complex size-, surface-, and composition-dependent catalytic properties and low active site density. In this work, we designed a single-atom Pt supported on Ni(OH) 2 nanoplates/nitrogen-doped graphene (Pt 1 /Ni(OH) 2 /NG) as the first example for constructing a single-atom catalyst based electrochemical nonenzymatic glucose sensor. The resulting Pt 1 /Ni(OH) 2 /NG exhibited a low anode peak potential of 0.48 V and high sensitivity of 220.75 μA mM –1 cm –2 toward glucose, which are 45 mV lower and 12 times higher than those of Ni(OH) 2, respectively. The catalyst also showed excellent selectivity for several important interferences, short response time of 4.6 s, and high stability over 4 weeks. Experimental and density functional theory (DFT) calculated results reveal that the improved performance of Pt 1 /Ni(OH) 2 /NG could be attributed to stronger binding strength of glucose on single-atom Pt active centers and their surrounding Ni…

Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas (CRISPR-associated) system has been widely explored for the detection of disease-related nucleic acids. Nevertheless, the simultaneous detection of multiple nucleic acids within one assay using the CRISPR-Cas system is still challenging. In this study, we develop an AND logic-gate-based CRISPR-Cas12a biosensing platform to achieve the sensitive colorimetric detection of dual miRNAs. Specifically, the DNA probe was designed to recognize the binary input of miRNAs and to output trigger DNA, which activated the CRISPR-Cas12a system to cut single-stranded DNA (ssDNA). The ssDNA on magnetic beads (MBs) was cleaved by the activated CRISPR-Cas12a, causing the separation of glucose oxidase (GOx) from MB and the subsequent generation of a colorimetric signal. The color change induced by 1 pM of target miRNAs can be directly distinguished by the naked eye and the instrumental limit of detection reaches 36.4 fM. The overexpressed miR-205 and miR-944 in the real human serum can be detected, allowing us to differentiate…

Defect engineering is an effective strategy to improve the catalytic activity of metal oxides, and quantitative characterization of surface defects is thus vital to the understanding and application of metal oxide catalysts. Herein, we found that ZnO nanoparticles with oxygen vacancy could trigger the luminol–H 2 O 2 system to emit a strong chemiluminescence (CL), and the CL intensity was strongly dependent on the oxygen vacancy of the ZnO nanoparticles. The mechanism of this CL reaction was discussed by means of the electron-spin resonance spectrum, X-ray photoelectron spectrum (XPS), and CL spectrum. The oxygen vacancy-dependent CL was attributed to the ability of the oxygen vacancy to readily adsorb and further dissociate H 2 O 2 into active • OH radicals. Taking advantage of this oxygen vacancy-dependent CL, we presented one method for quantifying the oxygen defects in ZnO. Compared with the current evaluation techniques (XPS and Raman spectroscopy), this CL method is rapid, low-cost, and easy to operate. This work introduces the CL technique into the field of material…

(Mi)RNAs are important biomarkers for cancers diagnosis and pandemic diseases, which require fast, ultrasensitive, and economical detection strategies to quantitatively detect exact (mi)RNAs expression levels. The novel coronavirus disease (SARS-CoV-2) has been breaking out globally, and RNA detection is the most effective way to identify the SARS-CoV-2 virus. Here, we developed an ultrasensitive poly-l-lysine (PLL)-functionalized graphene field-effect transistor (PGFET) biosensor for breast cancer miRNAs and viral RNA detection. PLL is functionalized on the channel surface of GFET to immobilize DNA probes by the electrostatic force. The results show that PGFET biosensors can achieve a (mi)RNA detection range of five orders with a detection limit of 1 fM and an entire detection time within 20 min using 2 μL of human serum and throat swab samples, which exhibits more than 113% enhancement in terms of sensitivity compared to that of GFET biosensors. The performance enhancement mechanisms of PGFET biosensors were comprehensively studied based on an electrical biosensor theoretical…

The successful application of electrochemiluminescence (ECL) in various fields required continuous exploration of novel ECL signal emitters. In this work, we have proposed a pristine ECL luminophor named NiRu MOFs, which owned extremely high and stable ECL transmission efficiency and was synthesized via a straightforward two-step hydrothermal pathway. The foundation framework of pure Ni-MOFs with the initial structure was layered-pillared constructed by the coordinated octahedrally divalent between nickel and terephthalic acid (BDC). The terephthalates were coordinated and pillared directly to the nickel hydroxide layers and the three-dimensional framework was formed, which had a weak ECL response strength. Then, the ruthenium pyridine complex was recombined with pure Ni-MOFs to produce NiRu MOFs and part of the introduced ruthenium was atomically dispersed in the layered-pillared structure through an ion-exchange method, which led to the ECL luminous efficiency being significantly boosted more than pure Ni-MOFs. In order to verify the superiority of this newly synthesized…

Antibiotic residues in the environment and in foods pose a serious threat to ecosystems and human health. Developing sensitive and on-site detection methods is therefore in high demand. In this work, a portable paper-based colorimetric sensor with a smartphone platform with an ultrahigh sensitivity has been designed for on-site and quantitative analysis of antibiotic residues based on aptamer-regulated nanozyme activity. The developed excellent peroxidase-like nanozymes, carbon-protected NiCo bimetal oxides with a unique hollow nanocage structure (NiCo@C HCs), could effectively catalyze the oxidation of chromogenic substrates by H 2 O 2 . Once bound to a specific aptamer, the enzyme-mimicking activity of NiCo@C HCs is obviously inhibited as a result of the masking of active sites but could be restored via the target-aptamer recognition. Herein, the aptamer-modified NiCo@C HCs are embedded on paper pieces to construct paper-based biochips for visual detection. Meanwhile, a smartphone platform is integrated for the signal readout. Using enrofloxacin (ENR) as an analyte model, the…

The functionalization of covalent organic frameworks (COFs) with biomacromolecules can extend their functions, which is the premise of their application in biomedical research. However, strategies to functionalize COFs with biomacromolecules, which can ensure the stability in complex medium and minimize the undesired effects, are still lacking. In this work, we have proposed a strategy to functionalize COFs with DNA by covalently linking DNA to the functional group on the COF surface through Cu(I)-catalyzed azide/alkyne cycloaddition (CuAAC) reaction. The as-prepared DNA-functionalized COFs (DNA-COFs) can exhibit good hybridization ability and cargo loading ability; thus, we have designed a DNA-COF-based nanoprobe and then fabricated an electrochemical biosensor for the detection of exosomes. In this design, the functionalization with DNA enables COFs to recognize and capture exosomes, and the encapsulation of a large number of methylene blue (MB) in COFs facilitates signal amplification, which can enhance the sensitivity of the biosensor. Moreover, by simply replacing the…

Toward the adoption of artificial intelligence-enabled wearable sensors interconnected with intelligent medical objects, this contactless multi-intelligent wearable technology provides a solution for healthcare to monitor hard-to-heal wounds and create optimal efficiencies for clinical professionals by minimizing the risk of disease infection. This article addressed a flexible artificial intelligence-guiding (FLEX-AI) wearable sensor that can be operated with a deep artificial neural network (deep ANN) algorithm for chronic wound monitoring via short-range communication toward a seamless, MXENE-attached, radio frequency-tuned, and wound dressing-integrated (SMART-WD) bandage. Based on a supervised training set of on-contact pH-responsive voltage output, the confusion matrix for healing-stage recognition from this deep ANN machine learning revealed an accuracy of 94.6% for the contactless measurement. The core analytical design of these smart bandages integrated wound dressing of poly(vinyl acrylic) gel@PANI/Cu 2 O NPs for instigating pH-responsive current during the wound healing…

Porous organic cages (POCs) are an emerging class of porous materials that have aroused considerable research interest because of their unique characteristics, including good solubility and a well-defined intrinsic cavity. However, there have so far been no reports of chiral POCs as chiral stationary phases (CSPs) for enantioseparation by high-performance liquid chromatography (HPLC). Herein, we report the first immobilization of a chiral POC, NC1-R, on thiol-functionalized silica using a mild thiol-ene click reaction to prepare novel CSPs for HPLC. Two CSPs (CSP-1 and CSP-2) with different spacers have been prepared. CSP-1, with a cationic imidazolium spacer, exhibited excellent enantioselectivity for the resolution of various racemates. Twenty-three and 12 racemic compounds or chiral drugs were well enantioseparated on the CSP-1-packed column under normal-phase and reversed-phase conditions, respectively, including alcohols, diols, esters, ethers, ketones, epoxides, organic acids, and amines. In contrast, chiral resolution using CSP-2 (without a cationic imidazolium spacer)-packed…

Mitochondria, as “cell energy stations”, are involved in the regulation of various cell functions. Recent investigations revealed that mitochondrial dysfunction that can cause an intracellular viscosity mutation, a process that is associated with an increasing number of diseases that are not curable or manageable. However, conventional viscometers cannot be used to monitor the viscosity changes in living cells and in vivo. In order to cater to the complex biological environment, we present a chemical toolbox, MI-BP-CC, that employs N, N -diethyl and double bonds as sensitive sites for viscosity based on the TICT mechanism (twisted intramolecular charge transfer) to monitor the viscosity of living cells and fatter liver mice. MI-BP-CC features good mitochondrial targeting and a near-infrared emission. Surprisingly, in the presence of viscosity, the MI-BP-CC probe exhibited an ultrasensitive model for viscosity detection showing a red fluorescence signal from a silent “off” state to “on”. More importantly, utilizing the satisfactory detection performance of MI-BP-CC, we have…

In this work, we synthesized a two-dimensional fluorescent covalent–organic framework (TFPB–TTA COF) nanosheet by selecting and designing reactive monomers to realize the dual-functional processing of nitrophenols. The staggered benzene ring, triazine structure, and imine bond (C═N) of the TFPB–TTA COF can capture free nitrophenols through hydrogen bonding and conjugation interaction, and then, the photoinduced electron transfer and fluorescence resonance energy transfer (FRET) between the TFPB–TTA COF and nitrophenols affects the fluorescence emission of the TFPB–TTA COF, realizing the fluorescence sensing of nitrophenols. The large K sv values and the low detection limit suggest that the TFPB–TTA COF can serve as sensitive and selective fluorescence sensors for nitrophenol detection in an aqueous system. At the same time, the strong interaction combined with the porous network structure of the TFPB–TTA COF facilitates the efficient adsorption and removal of nitrophenols. Especially for 2,4,6-trinitrophenol, the maximum adsorption capacity can reach 1045.53 mg/g with good…

Nonalcoholic fatty liver disease (NAFLD) is a global health issue. Peroxynitrite and liver viscosity have recently been found to be potential biomarkers of NAFLD. Therefore, it is of great significance to develop dual-response fluorescent probes for simultaneous detecting peroxynitrite and viscosity. We report herein a new probe (CQ) that can simultaneously detect peroxynitrite and viscosity at two independent fluorescent channels without signal crosstalk. CQ shows high selectivity, rapid response, good water solubility, low cytotoxicity, and mitochondrial localization properties. In particular, CQ responds sensitively to viscosity and peroxynitrite with off-on fluorescence changes at 710 and 505 nm, respectively. The wavelength gap between these two channels is more than 200 nm, ensuring that there is no signal crosstalk during detection. With this property, the probe was applied to simultaneously detect mitochondrial viscosity and peroxynitrite and image the changes of liver viscosity and peroxynitrite concentration during the pathogenesis of NAFLD. All results show that the CQ…

Simple and accurate monitoring of urinary dopamine (DA) concentration is significant, which is helpful for the assessment or exclusion of catecholamine-producing tumors, such as pheochromocytoma and paraganglioma. Herein, a fluorescence/colorimetry/smartphone triple-mode sensing platform for DA determination was constructed using copper ion (Cu 2+ )-modified hydrazone-linked covalent organic frameworks (Cu-BTA-COF). Cu-BTA-COF with 21.67 wt % copper content exhibited peroxidase-mimic activity. After adding H 2 O 2 and 1,3-dihydroxynaphthalene, the Cu-BTA-COF platform can sensitively and selectively detect DA in three modes with consistent results. In fluorescence/colorimetry/smartphone modes, the linear ranges of DA were 1–10, 0.2–40, and 1–10 μM, with related detection limits of 7.2, 8.6, and 23 nM, respectively. Moreover, the Cu-BTA-COF platform can be explored for DA determination in human urine samples with satisfactory recoveries (97.6–100.4%) in all the three modes, suggesting the potential practical application of the Cu-BTA-COF platform for DA detection in urine.

Graphdiyne (GDY) has been considered as an appealing electrode material for electrochemical sensing because of its alkyne-rich structure and high degrees of π-conjugation, which shows great affinity to heavy metal ions and pollutant molecules via d−π and π–π interactions. However, the low surface area and poor conductivity of bulk GDY limit its electrochemical performance. Herein, a two-dimensional ultrathin GDY/graphene (GDY/G) nanostructure was synthesized and used as an electrode material for electrochemical sensing. Graphene plays the role of an epitaxy template for few-layered GDY growth and conductive layers. The formed few-layered GDY with a high surface area possesses abundant affinity sites toward heavy metal ions (Cd 2+, Pb 2+ ) and toxic molecules, for example, nitrobenzene and 4-nitrophenol, via d−π and π–π interactions, respectively. Moreover, hemin as a key part of the enzyme catalytic motif was immobilized on GDY/G via π–π interactions. The artificial enzyme mimic hemin/GDY/G-modified electrode exhibited promising ascorbic acid and uric acid detection performance with…

Available automated methods for peak detection in untargeted metabolomics suffer from poor precision. We present NeatMS, which uses machine learning based on a convoluted neural network to reduce the number and fraction of false peaks. NeatMS comes with a pre-trained model representing expert knowledge in the differentiation of true chemical signal from noise. Furthermore, it provides all necessary functions to easily train new models or improve existing ones by transfer learning. Thus, the tool improves peak curation and contributes to the robust and scalable analysis of large-scale experiments. We show how to integrate it into different liquid chromatography-mass spectrometry (LC-MS) analysis workflows, quantify its performance, and compare it to various other approaches. NeatMS software is available as open source on github under permissive MIT license and is also provided as easy-to-install PyPi and Bioconda packages.

Ferroptosis is of great importance in physiological and pathological processes, which is associated with various inflammation-related diseases, cardiovascular diseases, and even cancer. Ferroptosis can cause abnormal change of reactive oxygen species (ROS) in mitochondria. Hypochlorous acid (HClO) acts as a typical ROS. Therefore, it is needed to study the relationship between mitochondrial morphology and HClO changes during ferroptosis at the subcellular level. To this end, a near-infrared-excitation/emission fluorescent probe, HD-Br-1, for rapid detection of mitochondrial HClO was developed based on the specific oxidative cleavage of the N, N -dimethylthiocarbamate moiety. The fluctuation in mitochondrial HClO content and the change in mitochondrial morphology during ferroptosis were monitored in real time by super-resolution imaging. In addition, HD-Br-1 was successfully applied to monitor exogenous and endogenous mitochondrial HClO during cell ferroptosis and visualize tumor to discriminate from healthy tissues. Therefore, we believe that HD-Br-1 could provide a valuable…

Mercury ion (Hg 2+ ) is one of the most harmful heavy metal ions with the greatest impact on public health. Herein, based on the excellent catalytic activity toward 3,3′,5,5′-tetramethylbenzidine (TMB) and the strong photocurrent-polarity-switching ability to SnS 2 photoanode of the split G-quadruplex–hemin complex, the magnetic NiCo 2 O 4 @SiO 2 –NH 2 sphere-assisted colorimetric and photoelectrochemical (PEC) dual-mode sensing platform was developed for the Hg 2+ assay. First, the amino-labelled single-stranded DNA1 (S1) was immobilized on NiCo 2 O 4 @SiO 2 –NH 2 and then partly hybridized with another single-stranded DNA2 (S2). When Hg 2+ was present, the thymine–Hg 2+ –thymine base pairs between S1 and S2 were formed, causing the formation of the split G-quadruplex in the presence of K + . After addition of hemin, the split G-quadruplex–hemin complex was obtained and effectually catalyzed the H 2 O 2 -mediated oxidation of TMB. Thus, the color and absorbance intensity of the TMB solution were changed, resulting in the visual and colorimetric detection of Hg 2+ . The linear…

An electrochemiluminescence (ECL) sensor provides a sensitive and convenient method for early diagnosis of diseases; however, it is still a challenge to develop simple and sensitive sensing platforms based on efficient ECL signals and luminophore groups. Porphyrin-based metal–organic frameworks (MOFs) show great potential in ECL sensing; however, the mechanism and structure–activity relationship, as well as application, are rarely reported. Herein, hydrothermal reactions obtained porphyrin Zr-MOFs (PCN-222) with different specific surface areas, pore sizes, structures, and surface charge states by tuning the reaction time were developed, which served both as the ECL luminophore, coreaction promoter for S 2 O 8 2–, and a connection in the ECL immunoassay. By progressively controlling the condition of the hydrothermal reaction, PCN-222 with large surface area-abundant micropores can be obtained, which has good conductivity and positively charged surfaces, obtaining excellent ECL performance. The ECL performance and the enhancement mechanism were investigated in detail. Using…

Analysis of cortisol levels in human sweat is increasingly important as it can be a “stress biomarker” in stress-related disorders, giving real-time information about human health status. In this study, a portable 3D microfluidic origami biosensor based on a smartphone was developed for cortisol-level detection in human sweat. Molybdenum disulfide (MoS 2 ) nanosheet-mediated fluorescence resonance energy transfer (FRET) and fluorescently labeled aptamers were employed in the biosensing process. A multilayer-structured 3D origami microfluidic chip was fabricated and functionalized to facilitate low-volume perspired human sweat collection, transportation, and detection. The translatability of the biosensor was exhibited by the fluorescence analysis in a smartphone mounted in a custom-designed holder. The critical design parameters of the microfluidic origami biosensor, including the characterization of various paper substrates, the concentration of MoS 2 nanosheets, and the incubation/reaction time, were adjusted to obtain an acceptable range for the assay dynamic range and limit of…

CRISPR-associated (Cas) protein systems have been increasingly incorporated in nucleic-acid diagnosis. CRISPR/Cas12a can cleave single-stranded DNA (ssDNA) after being guided to the target double-stranded DNA (dsDNA) with crRNA, making it a specific tool for dsDNA detection. Assisted by nucleic acid preamplification, CRISPR/Cas12a enables dsDNA detection at the attomolar level. However, such mandatory preamplification in CRISPR/Cas12a also accompanies the extra step of transferring preamplification products into the CRISPR/Cas12a system, which is not only cumbersome and time-consuming but also induces the risk of cross-contamination. Herein, we demonstrate a multiplex-crRNA strategy to enhance the sensitivity of the CRISPR/Cas12a system without any preamplification. This multiplex-crRNA strategy harnesses multiple sequences of crRNA which target different regions of the same dsDNA substrate in the same CRISPR/Cas12a system. Therefore, detection signals are accumulated without amplification, which augments the conventional detection limit. For application demonstration, the B646L…

We develop a novel noninvasive wearable biosensor to simultaneously assay multiple key biomarkers in human tears. The sensor is uniquely designed as a convenient-to-use eye patch that can be easily and comfortably fit under eyes. Different sensing regions on the eye patch are modified with specific chromogenic reagents to selectively determine the hydrogen ion (pH), protein, ascorbic acid, and glucose in tears. The assay takes only one drop of tear (∼20 μL) and a 30 s response time. The resulting color signals from each sensing region on the eye patch are captured and analyzed using a smartphone, and a cloud model based on the internet of things concept is recommended, which can extend the application of the eye patch biosensors in point-of-care testing and health monitoring, especially during the pandemic. Tear detection of the human subjects is performed using the proposed method, and the results display good correlation to parallel measurements using assay kits. The results show the great potential and versatility of the proposed eye patch biosensor for noninvasive determination…

Ultrasmall gold nanoclusters (AuNCs) are emerging as promising luminescent nanoprobes for bioimaging due to their fantastic photoluminescence (PL) and renal-clearable ability. However, it remains a great challenge to design them for in vivo sensitive molecular imaging in desired tissues. Herein, we have developed a strategy to tailor the PL and biofate of near-infrared II (NIR-II)-emitting AuNCs via ligand anchoring for improved bioimaging. By optimizing the ligand types in AuNCs and using Er 3+ -doped lanthanide (Ln) nanoparticles as models, core–satellite Ln@AuNCs assemblies were rationally constructed, which enabled 2.5-fold PL enhancement of AuNCs at 1100 nm and prolonged blood circulation compared to AuNCs. Significantly, Ln@AuNCs with dual intense NIR-II PL (from AuNCs and Er 3+ ) can effectively accumulate in the liver for ratiometric NIR-II imaging of H 2 S, facilitated by H 2 S-mediated selective PL quenching of AuNCs. We have then demonstrated the real-time imaging evaluation of liver delivery efficacy and dynamics of two H 2 S prodrugs. This shows a paradigm to visualize…

High sensitivity and specificity nucleic acid detection has been achieved by the Cas13a collateral effect in combination with a separate recombinase polymerase amplification (RPA). However, these emerging methods cannot provide accurate quantification of nucleic acids because the two-step assay performance may be compromised if the RPA and Cas13a reactions are simply unified in a single step. In this work, we first addressed the challenges associated with enzymatic incompatibility and the macromolecular crowding effect in the one-pot assay development, making the consolidated RPA-Cas13a assay a facile and robust diagnostic tool. Next, we found that the one-pot reaction cannot precisely quantify the targets at low concentrations. Thus, by leveraging droplet microfluidics, we converted the one-pot assay to a digital quantification format, termed Microfluidics-Enabled Digital Isothermal Cas13a Assay (MEDICA). Due to the droplet compartmentation, MEDICA greatly accelerates the reaction and enables relative detection in 10 min and the end-point quantification in 25 min. Moreover, MEDICA…

The electrochemiluminescence (ECL) immunoassay with its visual and high-throughput detection has received considerable attention in the past decade. However, the development of a facile and cost-effective ECL device is still a great challenge. Herein, a single-electrode electrochemical system (SEES) for the visual and high-throughput ECL immunoassay was developed. The SEES was designed by attaching a plastic sticker with multiple holes onto a single carbon ink screen-printed electrode based on a resistance-induced potential difference. Due to its excellent properties of adsorption and bioaffinity, the carbon ink screen-printed electrode is applied to immobilize antibodies. When cardiac troponin I (cTnI), a specific biomarker of acute myocardial infarction, is present, it will be captured by the immobilized cTnI antibodies on the electrode surface, inhibiting electron transfer, resulting in a decrease of the ECL intensity of the luminol–H 2 O 2 system. Using a smartphone as the detector, cTnI could be determined, ranging from 1 to 1000 ng mL –1, with a detection limit of 0.94 ng mL…

Circulating tumor cells (CTCs) are considered reliable cancer biomarkers for the liquid biopsy of many types of tumors. The direct detection of CTCs in human blood with normal biosensors, however, remains challenging because of severe biofouling in blood that contains various proteins and a large number of cells. Herein, we report the construction of an antifouling electrochemical biosensor capable of assaying CTCs directly in blood, based on a designed multifunctional peptide and the electrodeposited conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT). The designed peptide possesses antifouling capability in complex biological media and specific recognition ability to capture breast cancer cells MCF-7. Meanwhile, electrodeposited PEDOT can promote electron transfer at the sensing interface, improve the signal-to-noise ratio for the detection, and thus enhance the sensitivity of the biosensor. The integration of the multifunctional peptide and conducting polymer PEDOT ensures that the developed biosensor is able to perform directly in blood samples without purification or…

Skin interstitial fluid (ISF) containing a great variety of molecular biomarkers derived from cells and subcutaneous blood capillaries has recently emerged as a clinically potential component for early diagnosis of a wide range of diseases; however, the minimally invasive sampling and detection of cell-free biomarkers in ISF is still a key challenge. Herein, we developed microneedles (MNs) that consist of gelatin methacryloyl (GelMA) and graphene oxide (GO) for the enrichment and sensitive detection of multiple microRNA (miRNA) biomarkers from skin ISF. The GO-GelMA MNs exhibited robust mechanical properties, fast sampling kinetics, and large swelling capacity, which enabled collecting ISF volume high to 21.34 μL in 30 min, facilitating effective miRNA analysis. It preliminarily realized the sensitive detection of three types of psoriasis-related miRNAs biomarkers either on the patch itself or in solution after release from the hydrogel by combining catalytic hairpin assembly signal amplification reaction. The automated and minimally invasive ISF miRNA detection technology of…

In traditional lateral flow immunoassays (LFIA) for pathogens detection, capture antibody (CA) is necessary and usually conjugated to Au nanoparticles (NPs) in order to label the target analyte. However, the acquisition process of the Au–CA nanoprobe is relatively complicated and costly, which will limit the application of LFIA. Herein, p-mercaptophenylboronic acid-modified Au NPs (namely Au–PMBA nanocrabs), were synthesized and applied for a new CA-independent LFIA method. The stable Au–PMBA nanocrabs showed outstanding capability to capture both Gram-negative bacteria and Gram-positive bacteria through covalent bonding. The acquired Au–PMBA–bacteria complexes were dropped onto the strip, and then captured by the detection antibody on the test line (T-line). Take Escherichia coli O157:H7 as an example, the gray value of T-line was proportional to the bacteria concentration and the linear range was 10 3 –10 7 cfu·mL –1 . This CA-independent strategy exhibited higher sensitivity than the traditional CA-dependent double antibody sandwich method, because detection limit of the former…

Recently, single-atom catalysts (SACs) have been used to construct biosensors for the determination of organophosphorus pesticides (OPs). However, most nanozymes including SACs are peroxidase-like enzymes and require highly toxic and unstable hydrogen peroxide (H 2 O 2 ) as a co-reactant to generate reactive oxygen species. Inspired by the heme site of cytochrome c oxidases (C c os), the construction of Fe-N 5 -coordinated SACs by introducing axial N ligands is expected to bind O 2 to generate active metal–oxygen intermediates. Herein, a SAC with an Fe-N 5 active center confined by hierarchically porous carbon nanoframes (Fe SAs/N 5 -pC-4) was prepared by a polymerization–pyrolysis–evaporation–etching strategy, and its underlying enzyme-like mechanism was uncovered through experiments and density functional theory calculations. The 100% metal atom utilization, increased accessible active sites, accelerated mass transfer, excellent hydrophilicity, and an electron-driven mechanism of axial N endow the SAC with enhanced oxidase-like activity. Notably, its catalytic rate constant (0.398…

Enzymes featuring high catalytic efficiency and selectivity have been widely used as the sensing element in analytical chemistry. However, the structural fragility and poor machinability of an enzyme significantly limit its practicability in biosensors. Herein, we develop a robust and sensitive hybrid biosensor by means of co-encapsulating enzymes into a defective metal-organic framework (MOF), followed by a double-crosslinked alginate gelatinization. The defective MOF encapsulation can enhance the stability of enzymes, yet well preserve their biocatalytic function, while the alginate gelatinization allows the MOF biohybrid high stretchability and mechanical strength, which facilitates the integration of a bead-, fiber-, and sheet-like portable biosensor. In this work, the enzymes consisting of glucose oxidase and peroxidase are co-encapsulated into this MOF hydrogel, and it can efficiently convert glucose into a blue-violet product through the biocatalytic cascade of encapsulated enzymes, enabling the colorimetric biosensing of glucose on a miniaturized MOF hydrogel when coupling…

The self-luminescence behavior of lanthanide MOFs (Ln-MOFs) due to the unique antenna effect is considered to be a promising electrochemiluminescence (ECL) emission for biosensors. It is more challenging for Ln-MOFs on account of the difficulty to stimulate Ln ions with the desired energy-transfer efficiency to produce stronger ECL emissions at a low potential. Here, guided by a second ligand-assisted energy-transfer strategy, we present an efficient self-enhanced luminescence mixed-ligand Eu-MOF as an ECL signal probe for an oriented antibody-decorated biosensing platform with a low detection limit and a broad detection range. Diamino terephthalic acid (NH 2 –H 2 BDC) and 1,10-phenanthroline (Phen) were selected as the first and second ligands, respectively, to form highly conjugated structures, as well as suppress the nonradiative energy transfer. Impressively, Phen precisely adjusts the energy gap between the triplet ligand and the excited state of Eu 3+, realizing the self-enhancement of ECL efficiency of the Eu-MOF. The mixed ligand adjusted the molar ratio to obtain the stable…

Single-cell proteomics (SCP) has great potential to advance biomedical research and personalized medicine. The sensitivity of such measurements increases with low-flow separations (<100 nL/min) due to improved ionization efficiency, but the time required for sample loading, column washing, and regeneration in these systems can lead to low measurement throughput and inefficient utilization of the mass spectrometer. Herein, we developed a two-column liquid chromatography (LC) system that dramatically increases the throughput of label-free SCP using two parallel subsystems to multiplex sample loading, online desalting, analysis, and column regeneration. The integration of MS1-based feature matching increased proteome coverage when short LC gradients were used. The high-throughput LC system was reproducible between the columns, with a 4% difference in median peptide abundance and a median CV of 18% across 100 replicate analyses of a single-cell-sized peptide standard. An average of 621, 774, 952, and 1622 protein groups were identified with total analysis times of 7, 10, 15, and 30 min,…

Metal–organic frameworks (MOFs) are widely used to mimic enzymes for catalyzing chemical reactions; however, low enzyme activity limit their large-scale application. In this work, gold nanorods/metal–organic frameworks (Au NRs/Fe-MOF) hybrids were successfully synthesized for photo-enhanced peroxidase-like catalysis and surface-enhanced Raman spectroscopy (SERS). The enzyme-like activity of Au NRs/Fe-MOF hybrids was significantly enhanced under localized surface plasmon resonance (LSPR), because the hot electrons produced on Au NRs surface were transferred into Fe-MOF, activating the Fenton reaction by Fe 3+ /Fe 2+ conversion and preventing the recombination of hot electrons and holes. This photo-enhanced enzyme-like catalytic performance was investigated by X-ray photoelectron spectrometry (XPS), electrochemical analysis, activation energy measurement, and in situ Raman spectroscopy. Afterward, Methylene Blue (MB) was chosen to demonstrate the photo-enhanced peroxidase-like performance of Au NRs/Fe-MOFs. The Au NRs/Fe-MOF caused chemical and electromagnetic enhancement of Raman…

The innovative assembly of luminescent hydrogen-bonded organic frameworks (HOFs) into multifunctional optical sensors is of great significance for developing advanced materials. Herein, we report a facile room-temperature synthesis strategy for the luminol HOF modified by Tb 3+ (Lumi-HOF@Tb) and featuring sensitive chemiluminescence and fluorescence characteristics. Lumi-HOF@Tb is further pioneered as a dual-signal sensor for selective detection of α-glucosidase, a type of enzyme that plays a crucial role in the digestion of carbohydrates, and screening of its inhibitors. The sensor is constructed by combining the dual optical characteristics of luminol from the HOF and lanthanide ion assistance. From the hydrolysis of α-glucosidase and the 4-nitrophenyl-α- d -glucopyranoside (pNGP) substrate emerges the fluorescent luminol– p -nitrophenol (pNP) complex at 466 nm and changes the inner filter absorption to recover Tb 3+ characteristic fluorescence at 546 nm; luminol also produces a chemiluminescence signal driven by H 2 O 2 from additional glucose oxidase-catalyzed hydrolysis of α- d…

Meso-substituted boron dipyrromethenes (BODIPYs) provide a potential and innovative strategy for the synergistic construction of aggregation-induced emission (AIE) probes and fluorescent rotors for monitoring cellular viscosity changes, which play critical roles in understanding the function of viscosity in its closely associated diseases. Therefore, for the first time, a BODIPY-based fluorescent probe ( 1 ) with a rotatable meso- benzothiazole group was rationally designed and synthesized, showing both good viscosity-responsive and AIE properties. Probe 1 through direct linkage with the thiazole group, showed nearly no emission in low viscous solvents; however, a strong emission at 534 nm appeared and increased gradually with the increase in viscosity, attributing to the efficient restriction of the rotatable meso -benzothiazole group. The intensity (log I 534 ) displayed a good linear relationship with viscosity (log η) in the viscous range of 0.59–945 cP in methanol/glycerol mixtures. Interestingly, 1 showed enhanced emission at 534 nm in 70% water compared to pure acetonitrile…

Covalent organic frameworks (COFs) have attracted widespread attention in the electrochemiluminescence (ECL) field owing to their high load capacity of ECL luminophores and porous structures, but their ECL performance is still limited by the intrinsic poor conductivity (generally <10 –8 S m –1 ). To address this shortcoming, we used 2,3,6,7,10,11-hexaaminotriphenylene (HATP) and 2,3,6,7,10,11-hexahydroxytriphenylene (HHTP) to synthesize a conductive COF (HHTP-HATP-COF, conductivity = 3.11 × 10 –4 S m –1 ). Compared with HATP, HHTP, and low-conductive HHTP-DABZ-COF, HHTP-HATP-COF exhibited superior ECL performance, not only because HHTP-HATP-COF possessed massive ECL luminophores but also because its conductive porous framework accelerated charge transport in the whole framework and improved the utilization ratio of ECL luminophores. More interestingly, the ECL intensity of the HHTP-HATP-COF/S 2 O 8 2– system was further improved after pre-reduction electrolysis due to the accumulation of HHTP-HATP-COF cation radicals. The experimental results showed that the ECL intensity of the…

The development of modern technologies has acclimatized biosensors to complicated applicable scenarios with integrated properties as a whole instead of the pursuit of a single-point breakthrough. Here, we targeted a few concerns in the development of enzyme-based biosensors, including stability, analyte enrichment, and signal transduction, and developed a general biosensing model utilizing enzymes, aggregation-induced emission (AIE) luminogens, and stimuli-responsive framework materials as the units. We propose such proof-of-concept of glucose biosensors by coencapsulating glucose oxidase and AIE-type gold nanoclusters into acid-sensitive zeolite imidazolate framework (ZIF)-8 nanocrystals. The acid-activated degradation of ZIF-8 bridges the molecular signals produced by the enzyme-catalytic reaction of glucose and the photon signals generated by ZIF-8-induced AIE effects of gold nanoclusters, resulting in the "turn-off" model nanoprobes for glucose detection with high selectivity. After embedding the nanoprobes into hollow-out tapes, the formed paper biosensors can conveniently…

Energy deprivation and reduced levels of hydrogen sulfide (H 2 S) in the brain is closely associated with Alzheimer’s disease (AD). However, there is currently no fluorescent probe for precise exploration of both H 2 S and adenosine triphosphate (ATP) to directly demonstrate their relationship and their dynamic pattern changes. Herein, we developed a two-photon fluorescent probe, named AD-3, to simultaneously image endogenous H 2 S and ATP from two emission channels of fluorescent signals in live rat brains with AD. The probe achieved excellent selectivity and good detection linearity for H 2 S in the 0–100 μM concentration range and ATP in the 2–5 mM concentration range, respectively, with a detection limit of 0.19 μM for H 2 S and 0.01 mM for ATP. Fluorescence imaging in live cells reveals that such probe could successfully apply for simultaneous imaging and accurate quantification of H 2 S and ATP in neuronal cells. Further using real-time quantitative polymerase chain reaction and Western blots, we confirmed that H 2 S regulates ATP synthesis by acting on cytochrome C,…

Herein, a hand-drawing paper-based bipolar electrode (BPE) electrochemiluminescence (ECL) platform for M.SssI methyltransferase (M.SssI MTase) assay was proposed via employing high electrocatalytic Pt@CeO 2 as an ECL co-reaction accelerator and pencil-drawing graphite electric circuits as wires and electrodes. Notably, the introduction of pencil-drawing trace not only simplified the manufacturing process but also reduced the cost and saved fabricating time. Meanwhile, Pt@CeO 2 with good electrocatalytic activity and satisfactory chemical stability was used at the anode of the closed BPE-ECL device to accelerate the oxidation rate of uric acid. Due to the balanced charges of the bipolar electrode, the ECL response of the MnS: CdS@ZnS/S 2 O 8 2– system emitted on the cathode was enhanced. In situ growth of gold nanoparticles in the two electrode areas was convenient for DNA immobilization. With the above points in mind, the specific DNA double strands functionalized via Pt@CeO 2 were employed to identify M.SssI MTase. The unmethylated DNA double strands were cut by Hpa II…

A magnetic-assisted photoelectrochemical (PEC) and colorimetric (CL) dual-modal biosensing platform with high precision was established to monitor prostate-specific antigen (PSA) based on Bi 2 MoO 6 nanosheets (BMO) by coupling the aptamer-guided hybridization chain reaction (HCR) with the hydrolysate-induced vulcanization reaction of Bi 2 MoO 6 nanosheets. Upon addition of PSA, trigger DNA (tDNA) was released by the interaction between the target analyte and the aptamer and then further hybridized with anchor DNA (aDNA) conjugated on magnetic beads (MBs). The as-released tDNA initiated the target-assisted HCR in the presence of two alternating hairpin sequences (Bio-H1 and Bio-H2) to produce nicked long double-stranded DNA on the surface of MBs, where numerous alkaline phosphatase (ALP) enzymes could assemble with MBs through the biotin–avidin reaction, resulting in the hydrolysis of sodium thiophosphate (TP) to H 2 S. The as-produced H 2 S reacted with BMO to form vulcanized BMO (BMO-S), thus leading to obvious enhanced PEC performance under visible light with the color change…

Cancer is a health threat worldwide, and it is urgent to develop more sensitive cancer detection methods. Herein, a polarity-sensitive cell membrane probe (named COP ) was developed for detecting cancer cells and tumors sensitively and selectively at the cell membrane level. The probe shows a strong polarity-dependent fluorescence and excellent cell membrane targeting ability to visualize cell membrane with red fluorescence with a non-washing process. Notably, COP can selectively light up the tumor cell membranes, which reveals that cancer cell membranes have lower polarity than normal cell membranes. The giant unilamellar vesicle model and cell imaging studies proved this. Moreover, COP can effectively and selectively light up tumors. Overall, this work demonstrates that the polarity of the tumor cell membrane is quite different to normal cell membranes, and based on this, sensitive membrane probes can be developed to selectively visualize cancer cells and tumors, which opens up a new way for tumor diagnosis at the cellular level.

By combining the distinctive noninvasive feature with the peculiar complete functional implementation trait, fully integrated raw noninvasive biofluid glucose biosensors offer active and remote glucose monitoring while posing minimal harm or infection risks compared to the traditional invasive manner. However, each previously reported fully integrated raw noninvasive biofluid glucose biosensor is solely focused on single-type raw noninvasive biofluid analysis. Given the diversity and complexity of subjects’ physical conditions, single-type raw noninvasive biofluids are inappropriate to all crowds (e.g., sweat collection/analysis could be inapplicable for dermatopathic subjects). Here, we demonstrate the first example of a universal fully integrated nanoelectronic system with the unique capability to point-of-care and universally monitor diverse raw noninvasive biofluid (i.e., sweat, tears, and saliva) glucose by combining a flexible and disposable microfluidic enzymatic biosensor (named i ez Slice) for raw biofluid pump-free sampling and measurement with a customized, handheld, and…

The necessary step of directly adding hydrogen peroxide (H 2 O 2 ) into the detection system in traditional immunoassays hampers their applications as a portable device for point-of-care analysis due to the unstable liquid form of H 2 O 2 . Herein, a strategy of self-supplying H 2 O 2 and signal amplification triggering by copper peroxide nanodots encapsulated (CPNs) in metal–organic frameworks (ZIF-8) was proposed in an immunoassay for dual-signal detection of bisphenol A (a typical emerging organic pollutant), which was further fabricated as a lab-in-a-tube device integrated with a smartphone sensing platform. Herein, CPNs@ZIF-8 was modified on the antibody against bisphenol A; after the competitive binding of analytes, coating antigens, and antibodies, the released H 2 O 2 and Cu 2+ from encapsulated CPNs under the acidic condition will trigger a Fenton-like reaction to generate ·OH for oxidization of TMB; meanwhile, Cu 2+ could quench the fluorescence of GSH-Au NCs, resulting in dual-mode signals for measurements. Most importantly, self-supplying H 2 O 2 with high stability was…

Herein, we revealed the factors that affect the emission in mixed-ligand metal–organic frameworks (MOFs) with the combination of terephthalic acid (BDC), 2-aminoterephthalic acid (BDC-NH 2 ), and 2,5-dihydroxylterephthalic acid [BDC-(OH) 2 ] as models. The −NH 2 and −(OH) 2 groups change the π-conjugation and luminescence behaviors than BDC, so the ligands show different optical behaviors. The Zn 2+ ion with a 3d 10 full electronic structure shows little effect on the emission of the ligand and is selected as the metal node. We found that the emission of BDC is weak and incompatible to that of BDC-NH 2, so only the emission of BDC-NH 2 was observed in the BDC/BDC-NH 2 -MOF. Crosstalk occurs between the emissions from BDC and BDC-(OH) 2 for the single emission from BDC/BDC-(OH) 2 -MOFs, even different ratios are selected. The MOFs prepared with BDC-NH 2 and BDC-(OH) 2 show dual emission at 450 and 550 nm, while the relative intensity was easily tuned with the ligand ratio and excitation wavelength. Thus, abundant optical behaviors and extensive applications were realized, including…

A novel high-throughput aptamer microarray fluorescent method based on thioflavin T (ThT) was established for the sensitive detection of phoxim, parathion, fensulfothion, and isocarbophos. In this work, the aptamers in binding buffer tended to have the antiparallel G-quadruplex structure, which can bind ThT and release its potential fluorescence signal. However, when the organophosphorus pesticides (OPs) were present, partial aptamers preferred to bind them, forcing the displacement of ThT from the G-quadruplex and resulting in the significant decrease in fluorescence signal. Under optimal experimental conditions (12T spacer, 300 nM aptamer, and 80 μM ThT), the OP aptamer microarray has low limits of detection of 25.4 ng/mL for phoxim, 12.0 ng/mL for parathion, 7.7 ng/mL for fensulfothion, and 9.9 ng/mL for isocarbophos. The accuracy and reliability of the method is further verified by testing the recovery rate of OPs spiked in two different complicated sample matrices (pears and radishes). It is worth mentioning that not only the developed aptamer microarray technology has low…

The future of personalized diagnostics and treatment of cardiovascular diseases lies in the use of portable sensors. Portable sensors can acquire biomarker information in biological fluids such as sweat, an approach that mitigates the shortcomings of conventional hospital-centered healthcare. Low sensitivity, selectivity, and specificity remain bottlenecks for the widespread use of portable sensors. Herein, we demonstrate a portable sensor that simultaneously detects Na +, ascorbic acid, and human neuropeptide Y in sweat, all useful biomarkers to index cardiovascular health. The portable sensor comprises a six-electrode system containing three working electrodes, two reference electrodes, and one counter electrode. The working electrodes were prepared by depositing sensing components on carbon quantum dot (CQD) electrodes. The sensing mechanisms were based on selective ion recognition, enzyme catalytic reaction, and immune response, which guarantees specificity to corresponding targets. The CQDs offer massive reactive sites and effectively reduce the interfacial impedance during the…

Lipid droplets (LDs), which are ubiquitous organelles existing in almost all eukaryotic cells, have attracted a lot of attention in the field of cell biology over the last decade. For the biological study of LDs via fluorescence imaging, the superior LD fluorescent probes with environmental polarity-sensitive character are highly desired and powerful but are very scarce. Herein, we have newly developed such a kind of fluorescent probe named LDs-Red which enables us to visualize LDs and to further reveal their polarity information. This fluorescent probe displays the advantages of intense red/near-infrared emission, high LD staining specificity, and good photostability; thus, it would be very useful for LD fluorescence imaging application. As a result, the three-dimensional confocal imaging to visualize spatial distribution of LDs and the multicolor confocal imaging to simultaneously observe LDs and other cellular organelles have been realized using this new LD fluorescent probe. Furthermore, the polarity-sensitive emission character of this probe enables us to quantitatively…

Since aggregation-induced electrochemiluminescence (AIECL) combined the merits of aggregation-induced emission (AIE) and electrochemiluminescence (ECL), it has become a research hotspot recently. Herein, novel kinds of functional metal-organic frameworks (MOFs) with strong AIECL were reported through doping tetraphenylethylene (TPE) into UiO-66. Due to the porosity and highly ordered topological structure that caused the confinement effect of MOFs, the molecular motion of TPE was effectively limited within UiO-66, resulting in strong AIE. Meanwhile, the large specific surface area and porous structure of UiO-66 allowed TPE to react with coreactants more effectively, which was beneficial to ECL. Thus, the TPE-functionalized UiO-66 (TPE-UiO-66) showed excellent AIECL performance surprisingly. Inspired by this, a multiple convertible ECL resonance energy transfer (ECL-RET) system was constructed through a DNA Y structure that regulated the distance between the energy donor (TPE-UiO-66) and different energy acceptors (gold nanoparticles and Adriamycin). Furthermore, an ultrasensitive…

Exploring new electrochemiluminescence (ECL) luminophores with strong ECL emission is highly desirable for developing ultrasensitive ECL sensors. Herein, a pyrene-based hydrogen-bonded organic framework (Py-HOF) featuring prominent ECL performance was prepared by utilizing 1,3,6,8-tetrakis( p -benzoic acid) pyrene (H 4 TBAPy) with an aggregation-induced enhanced emission (AIEE) property as a building block, exhibiting a stronger ECL emission than those of H 4 TBAPy monomers, H 4 TBAPy aggregates, the low-porosity Py-HOF-210 °C and Py-HOF-180 °C. We have coined the term “the porosity- and aggregation-induced enhanced ECL (PAIE-ECL)” for this intriguing phenomenon. The Py-HOF displayed superb and stable ECL intensity, not only because the luminophore H 4 TBAPy was assembled into the Py-HOF via four pairs of O–H···O hydrogen bonds, which constrained the intramolecular movements to reduce nonradiative transition, but also because the H 4 TBAPy in Py-HOF was stacked in a slipped face-to-face mode to form J-aggregates that benefited the ECL enhancement. Furthermore, the high porosity of…

Exploiting low cost, water-soluble, and near-infrared (NIR) emissive electrochemiluminophores (ECLphores) is significantly important for biological applications. In this study, bright and NIR electrogenerated chemiluminescence (ECL) emissive copper nanoclusters (Cu NCs) were synthesized through a facile one-pot wet chemical reduction method. ECL properties of obtained Cu NCs were examined in the presence of potassium persulfate, resulting in maximum intensity at 735 nm, at least 135 nm red-shifted with respect to all other Cu NCs. Electrochemistry, photoluminescence (PL), and spooling ECL spectroscopies were used to track NIR ECL emission of Cu NCs ascribed to the monomeric excited states. Due to the abundant binding sites of bovine serum albumin (BSA) to anchor target biomolecules, a sandwich-type ECL immunosensor was thus fabricated using such BSA-templated Cu NCs as tags and alpha fetoprotein antigen (AFP) as a model protein for the first time. Without assisting any signal amplification strategies, the proposed NIR ECL biosensor exhibited a wide linear range (1–400 ng mL –1 ) and…

Raman spectroscopy is commonly used in microplastics identification, but equipment variations yield inconsistent data structures that disrupt the development of communal analytical tools. We report a strategy to overcome the issue using a database of high-resolution, full-window Raman spectra. This approach enables customizable analytical tools to be easily created─a feature we demonstrate by creating machine-learning classification models using open-source random-forest, K-nearest neighbors, and multi-layer perceptron algorithms. These models yield >95% classification accuracy when trained on spectroscopic data with spectroscopic data downgraded to 1, 2, 4, or 8 cm –1 spacings in Raman shift. The accuracy can be maintained even in non-ideal conditions, such as with spectroscopic sampling rates of 1 kHz and when microplastic particles are outside the focal plane of the laser. This approach enables the creation of classification models that are robust and adaptable to varied spectrometer setups and experimental needs.

In this work, an antigen-down photoelectrochemical (PEC) immunosensor based on a signal polarity switching strategy for the detection of cytokeratin 19 fragment 21-1 (CYFRA21-1) was proposed. 3,4,9,10-Perylene tetracarboxylic acid (PTCA) is a conjugated organic dye containing five benzene nuclei, which has excellent film-forming and optical properties. PTCA sensitized by SnS 2 can further improve the basal signal and the stability of the PEC immunosensor. Moreover, avidin-functionalized CuInS 2 as a signal probe can convert the basal anodic photocurrent to a cathodic photocurrent. Therefore, the PEC sensor realized the photocurrent polarity conversion before and after labeling. With avidin-functionalized CuInS 2, the polarity of the photocurrent was changed once CYFRA21-1 was detected. Therefore, the PEC immunosensor owns high sensitivity. The linear range of the immunosensor for the detection of CYFRA21-1 is 0.00001–500 ng·mL –1, and the detection limit is 3.5 fg·mL –1 . The PEC immunosensor has good stability, high selectivity, and good repeatability. This work may provide a new…

As emerging contaminants, microplastics are challenging to characterize, particularly when their size is at the nanoscale. While imaging technology has received increasing attention recently, such as Raman imaging, decoding the scanning spectrum matrix can be difficult to achieve result digitally and automatically via software and usually requires the involvement of personal experience and expertise. Herewith, we show a dual-principal component analysis (PCA) approach, where (i) the first round of PCA analysis focuses on the raw spectrum data from the Raman scanning matrix and generates two new matrices, with one containing the spectrum profile to yield the PCA spectrum and the other containing the PCA intensity to be mapped as an image; (ii) the second round of PCA analysis merges the spectrum from the first round of PCA with the standard spectra of eight common plastics, to generate a correlation matrix. From the correlation value, we can digitally assign the principal components from the first round of PCA analysis to the plastics toward imaging, akin to dataset indexing. We also…

Gastric cancer is a malignant tumor, and its early diagnosis remains challenging due to the lack of simple and sensitive detection methods and specific biomarkers. In this work, to improve the detection reliability, we developed a dual-mode detection strategy for the detection of two biomarkers associated with it. First, an N- and S-doped carbon dots-N-rich porous carbon nanoenzyme (N/ [email protected] ) was prepared by a two-step pyrolysis of thiourea-penetrated zinc-based zeolite imidazole framework. It was then combined with the 3,3′,5,5′-tetramethylbenzidine–H 2 O 2 system for the colorimetric detection of d -amino acids (i.e., d -proline ( d -Pro) and d -alanine ( d -Ala)) in saliva, based on d -amino acid oxidase catalyzing d -amino acid oxidation to produce H 2 O 2 . In this way, the low detection limits (S/N = 3) of d -Pro and d -Ala were 0.14 and 0.35 μM, respectively. Furthermore, N/ [email protected] was combined with the luminol–H 2 O 2 electrochemiluminescence (ECL) system and magnetic immune accumulation/separation strategy to detect the carcinoembryonic antigen (CEA)…

Phenol and its derivatives, known as persistent organic pollutants, have long threatened human health and environmental safety. There is an urgent need to develop convenient, low-cost, and multiplex analytical methods. Since phenols are substrates of laccase, they can be detected via laccase-catalyzed colorimetric assays. Nevertheless, the laccase-based assays cannot distinguish different phenols. Moreover, natural laccases suffer from high cost and low stability issues. To meet these needs, here we developed a laccase-like nanozyme sensor array for phenol detection and differentiation, which takes advantage of both nanozymes and cross-reactive sensor arrays. First, we examined a series of spinel-type transition metal oxides and found that manganese on octahedral sites profoundly affects the laccase-like activity of the materials. Based on the developed manganese-based spinel oxides (i.e., Mn 3 O 4, Zn 0.4 Li 0.6 Mn 2 O 4, and LiMn 2 O 4 ), a colorimetric sensor array was constructed. The sensor array could effectively identify and discriminate phenol and its derivatives and showed…

Improving the sensitivity of electrochemiluminescence (ECL) systems is highly desired for in vitro ECL diagnosis and bio-detections due to the often-low content of biomarkers in diseases. And dissolved O 2 (DO) as a co-reactant is considered superior to H 2 O 2 in the most commonly used luminol ECL systems due to better stability and low biotoxicity, but it still suffers from low ECL performance due to the low reactivity of DO. In this study, an efficient luminol-DO ECL system was developed through the complexing of Fe, Co dual single-atom catalysts (D-SACs) supported by N-doped graphene with the luminol-capped Ag nanoparticles (AgNPs). Benefiting from the electronic interaction between Fe and Co metal sites in the relevant D-SACs and plasmon enhancement of AgNPs, the performance of the corresponding ECL system could be significantly boosted up to ≈677-fold under optimal testing conditions, comparable to the classic luminol-O 2 system. Furthermore, the developed luminol-DO ECL system was successfully applied for the stable ultrasensitive detection of prostate-specific antigen (PSA)…

Eliminating false positives or negatives in analysis has been a challenge. Herein, a phenomenon of polarity-switching photocurrent of AgInS 2 quantum dot (QD)–DNA nanowires reversing nitrogen-doped porous carbon–ZnO (NPC–ZnO) nanopolyhedra was found for the first time, and a versatile photoelectrochemical (PEC) biosensor with a reversed signal was innovatively proposed for dual-target detection. NPC–ZnO is a photoactive material with excellent PEC properties, while AgInS 2 QDs as a photosensitive material match NPC–ZnO in the energy level, which not only promotes the transfer of photogenerated carriers but also switches the direction of PEC current. Furthermore, in order to prevent spontaneous agglomeration of AgInS 2 (AIS) QDs and improve its utilization rate, a new multiple-branched DNA nanowire was specially designed to assemble AgInS 2 QDs for constructing amplified signal probes, which not only greatly increased the load of AgInS 2 QDs but also further enhanced the photoelectric signal. When the target Hg 2+ -induced cyclic amplification process generated abundant RDNA, the DNA…

Nanozymes are a kind of nanomaterial mimicking enzyme catalytic activity, which has aroused extensive interest in the fields of biosensors, biomedicine, and climate and ecosystems management. However, due to the complexity of structures and composition of nanozymes, atomic scale active centers have been extensively investigated, which helps with in-depth understanding of the nature of the biocatalysis. Single atom nanozymes (SANs) cannot only significantly enhance the activity of nanozymes but also effectively improve the selectivity of nanozymes owing to the characteristics of simple and adjustable coordination environment and have been becoming the brightest star in the nanozyme spectrum. The SANs based sensors have also been widely investigated due to their definite structural features, which can be helpful to study the catalytic mechanism and provide ways to improve catalytic activity. This perspective presents a comprehensive understanding on the advances and challenges on SANs based sensors. The catalytic mechanisms of SANs and then the sensing application from the…

Drug-induced liver injury (DILI) is the most common clinical adverse drug reaction, which is closely associated with the oxidative stress caused by overproduced reactive oxygen species. Hepatic H 2 O 2, as an important biomarker of DILI, plays a crucial role in the progression of DILI. However, there remains a challenge to develop H 2 O 2 -activatable second near-infrared (NIR-II, 1000–1700 nm) small molecular probes with both a large Stokes shift and a long emission wavelength beyond 950 nm. Herein, we developed an activatable NIR-II fluorescent probe ( IR-990 ) with an acceptor-π-acceptor (A-π-A) skeleton for real-time detection of H 2 O 2 in vivo. In the presence of H 2 O 2, nonfluorescent probe IR-990 was successfully unlocked by generating a donor-π-acceptor (D-π-A) structure and switched on intense NIR-II fluorescence, exhibiting a peak emission wavelength at 990 nm and a large Stokes shift of 200 nm. Moreover, it was able to detect H 2 O 2 with high sensitivity and selectivity in vitro (LOD = 0.59 μM) and monitor the behavior of endogenous H 2 O 2 in the HepG2 cell model of…

The simultaneous analysis of the levels of multiple microRNAs (miRNAs) is critical to the early diagnosis of cancer. However, this analysis is challenging because of the low concentrations of miRNAs and their high sequence homology. Here, we report a general and programmable diagnostic strategy for miRNA analysis: Thermus thermophilus Argonaute (TtAgo)-assisted exponential isothermal amplification for multiplex detection (TEAM). This system combines exponential isothermal amplification (EXPAR), for target amplification, with programmable TtAgo cleavage, for the generation of the reporting signal. The TEAM assay achieved attomolar sensitivity with a rapid turnaround time (30–35 min). Because of the single-nucleotide precision of TtAgo, the system demonstrated robust multiplex capability in the simultaneous detection of four miRNA targets and the classification of let-7 family members. The TEAM assay was superior in differentiating colorectal cancer patients from healthy individuals relative to the conventional EXPAR and reverse transcription polymerase chain reaction (RT-PCR)…

Large RNA including mRNA (mRNA) has emerged as an important new class of therapeutics. Recently, this has been demonstrated by two highly efficacious vaccines based on mRNA sequences encoding for a modified version of the SARS-CoV-2 spike protein. There is currently significant demand for the development of new and improved analytical methods for the characterization of large RNA including mRNA therapeutics. In this study, we have developed an automated, high-throughput workflow for the rapid characterization and direct sequence mapping of large RNA and mRNA therapeutics. Partial RNase digestions using RNase T1 immobilized on magnetic particles were performed in conjunction with high-resolution liquid chromatography-mass spectrometry analysis. Sequence mapping was performed using automated oligoribonucleotide annotation and identifications based on MS/MS spectra. Using this approach, a >80% sequence of coverage of a range of large RNAs and mRNA therapeutics including the SARS-CoV-2 spike protein was obtained in a single analysis. The analytical workflow, including automated sample…

Lipid droplets (LDs) are involved in various physiological processes and associated with cancer development, and are regarded as a potential tumor marker for cancer diagnosis. Monitoring LDs is of prior importance to understand their involvement in biological mechanisms and the early detection of cancers. Highly sensitive and specific noninvasive fluorescent probes are particularly desirable for imaging LDs and cancer diagnosis. Herein, according to the high-viscosity and low-polarity microenvironment in LDs, we developed four easily prepared LDs-specific probes based on noncharged merocyanines. Among them, LD-1 absorbs and emits in the near-infrared (NIR) region with a large Stokes shift. Importantly, LD-1 displayed high sensitivity to high viscosity and low polarity, which allowed it to show high LDs-targeting ability. In cell imaging, LD-1 successfully probed the changes in LDs in the presence of oleic acid or during ferroptosis and was used to distinguish cancer cells from normal cells.

The discrimination between dead and live cells is crucial for cell viability evaluation. Carbon dots (CDs), with advantages like simple and cost-effective synthesis, excellent biocompatibility, and high photostability, have shown potential for realizing selective live/dead cell staining. However, most of the developed CDs with the live/dead cell discrimination capacity usually have low photoluminescence quantum yields (PLQYs) and excitation wavelength-dependent fluorescence emission (which can cause fluorescence overlap with other fluorescent probes and make dual-color live/dead staining impossible), and hence, developing ultrabright CDs with excitation wavelength-independent fluorescence emission property for live/dead cell discrimination becomes an important task. Here, using a one-pot hydrothermal method, we prepared ultrasmall (∼1.6 nm), ultrabright (PLQY: ∼78%), and excitation wavelength-independent sulfur-doped carbon dots (termed S-CDs) using rose bengal and 1,4-dimercaptobenzene as raw materials and demonstrated that the S-CDs could rapidly (∼5 min) and accurately…

With the growing consumption of caffeine-containing beverages, detection of caffeine has become an important biomedical, bioanalytical, and environmental topic. We herein isolated four high-quality aptamers for caffeine with dissociation constants ranging from 2.2 to 14.6 μM as characterized using isothermal titration calorimetry. Different binding patterns were obtained for the three single demethylated analogues: theobromine, theophylline, and paraxanthine, highlighting the effect of the molecular symmetry of the arrangement of the three methyl groups in caffeine. A structure-switching fluorescent sensor was designed showing a detection limit of 1.2 μM caffeine, which reflected the labeled caffeine concentration within 6.1% difference for eight commercial beverages. In 20% human serum, a detection limit of 4.0 μM caffeine was achieved. With the four aptamer sensors forming an array, caffeine and the three analogues were well separated from nine other closely related molecules.

This work proposed a novel double-engine powered paper photoelectrochemical (PEC) biosensor based on an anode–cathode cooperative amplification strategy and various signal enhancement mechanisms, which realized the monitoring of multiple miRNAs (such as miRNA-141 and miRNA-21). Specifically, C 3 N 4 quantum dots (QDs) sensitized ZnO nanostars and BiOI nanospheres simultaneously to construct a composite photoelectric layer that amplified the original photocurrent of the photoanode and photocathode, respectively. Through the independent design and partition of a flexible paper chip to functionalize injection holes and electrode areas, the bipolar combination completed the secondary upgrade of signals, which also provided biological reaction sites for multitarget detection. With the synergistic participation of a three-dimensional (3D) DNA nanomachine and programmable CRISPR/Cas12a shearing tool, C 3 N 4 QDs lost their attachment away from the electrode surface to quench the signal. Moreover, electrode zoning significantly reduced the spatial cross talk of related substances for…

A photoactive molecularly imprinted poly(2,5-benzimidazole)-modified vanadium dioxide–cupric tungstate (VO 2 –CuWO 4 ) as an efficient photosensitive n – n type-II heterojunction thin film was electrochemically deposited on Ti substrate for the selective and robust photoelectrochemical (PEC) bioanalysis of dopamine (DA). The optical absorption of n -VO 2 / n -CuWO 4 type-II heterojunction was capably broadened toward the visible region, which permitted superior light-harvesting and robust carriers generation, separation, and transfer processes significantly enhancing the anodic photocurrent, as confirmed by a series of PEC analyses. Findings revealed that the as-prepared label-free MIP-PEC sensor can quantitatively monitor DA in a linear range of 1 nM to 200 μM with a detection limit of 0.15 nM. This MIP-PEC sensor showed robust selectivity under conditions with high concentrations of interfering substances, which can be recovered in the real samples of urine, cocoa chocolate, and diluted yogurt, indicating its promising potential applications in biological and food samples. This…

There is a great demand to develop high-performance hydrogen sulfide (H 2 S) sensors, especially at the trace level for environmental protection, human healthcare, and food freshness monitoring. To this end, cellulose nanofiber (CNF)-templated CuO (CuO-C) was decorated with tungsten disulfide (WS 2 ) nanosheets (W-Cu-C) as the sensing layer of chemiresistive microelectromechanical system (MEMS) sensors for H 2 S recognition in this work. As compared to pristine CuO counterparts at 160.5 °C, the as-prepared W-Cu-C-10 sensors delivered a 30-fold enhanced response of 37 toward 0.5 ppm H 2 S at a lower optimal operation temperature of 100.1 °C. Moreover, a fast response/recovery speed of 37.2/33.9 s toward 0.5 ppm H 2 S and excellent long-term stability and selectivity were achieved. Compared with existing research and commercial products, the W-CU-C-10 sensors exhibited the remarkable superiorities of high sensitivity, the lowest detection limit of 200 ppb, and ultralow power consumption (8 mW). Also, the sensor showcased a nice on-site application potential for evaluating eggs’…