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The polarity and viscosity of the microenvironment are associated with the control of the onset and progression of pathological diseases, including inflammation, immuno-suppression and cancer. If appropriate treatment is neglected, alcoholic acute liver injury (AALI), the initial sign of alcoholic liver diseases, may transform into hepatic lesions. Therefore, it's crucial to create a particular probe to detect AALI swiftly and track its progression. Herein a polarity and viscosity dual-responsive crimson fluorescent probe (PPBI) was designed and developed, which can target mitochondria and lipid droplets. PPBI possesses aggregation-induced emission properties, good photostability and strong anti-interference ability against pH, metal ions, anions and biomolecules. This probe can distinguish cancer cells from normal ones using changes of green and red fluorescence, as well as identify changes in the cellular microenvironment associated with inflammatory and ferroptosis processes. In addition, changes in polarity and viscosity can be amplified by in vivo imaging in a mouse model to monitor alcohol-induced acute liver injury and to effectively detect the course of pharmacological intervention therapy. All the results suggest that PPBI could be a promising real-time fluorescence imaging tool for diagnosis and treatment of acute alcoholic liver injury. [Display omitted] [ABSTRACT FROM AUTHOR]

Alternariol (AOH) is one of the common mycotoxins existing in a variety of foods at low level. Aptamers hold great promise for the development of sensitive and rapid aptasensors, but suffer from the excessive length and the difficulty in identification of critical binding domains (CBDs). In this study, the 5 nt CBD of the original 59-nt AOH aptamer (AOH-59, K D = 423 nM) was identified to be a 'C' bulge in between two A-T base pairs. AOH-59 was successfully shortened to a 23 nt aptamer (AOH 6C, K D = 701 nM). A 30 nt bivalent aptamer B-2-3 (K D = 445 nM) and a 39 nt trivalent aptamer T-2-3 (K D = 274 nM) were obtained by simply incorporating one or two CBDs into AOH 6C. The AOH 6C-, B-2-3-, and T-2-3-based optical waveguide aptasensors possessed the unprecedented detection of limits (LODs, S/N = 3) of 42 ± 3, 6 ± 1 and 2 ± 1 fM, respectively. Using the AOH 6C-based sensor as an example, we further demonstrated the detection of AOH spiked in wheat powder with a LOD of 37 pg/g, 20-230-fold lower than those achieved by ELISAs. The sensor was capable for 35 times 2-min regeneration and the assay time including the extraction of AOH from wheat was only about 1 h. We not only devised the first aptasensors for AOH detection, but also provided a simple strategy to design multivalent aptamers for small molecule targets. [Display omitted] • The first DNA aptamer for alternariol with nM dissociation constant. • Identification of the 5 nt critical binding domain. • Bivalent and trivalent alternariol aptamers with improved affinity and short length. • Waveguide aptasensors for detection of alternariol in wheat with fM LODs. • Up to 35 times sensor regeneration. [ABSTRACT FROM AUTHOR]

Metastasis is a leading cause of cancer-related deaths. Hence, the discovery of more reliable metastasis-related biomarkers is crucial to improve the survival rate of cancer patients. W3 is an aptamer previously produced by the subtractive cell-SELEX using metastatic colorectal cancer cells as target cells and non-metastatic cells as negative cells. In this study, we aimed to evaluate whether the target molecule of W3 can potentially act as a metastatic biomarker. First, we obtained two cell subpopulations with different expression levels of the target molecule by W3-based cell sorting. Subsequently, we demonstrated that W3high cells have a higher metastatic potential than W3low cells both in vitro and in vivo. Further, immunohistochemical analysis revealed that W3 target expression is positively associated with metastasis and poor prognosis of CRC patients. Using mass spectrometry (MS) combined with pull-down, we identified that Ephrin type-A receptor 2 (EphA2) is the target of W3. EphA2's potential as a metastatic predictor was demonstrated by capturing W3-positive circulating tumor cells from CRC patients using a W3 probe. Based on these results, we put forward a stratagem for cell-SELEX-based biomarker discovery: selecting an aptamer through subtractive cell-SELEX towards the phenotype of interest; evaluating the functional phenotype of the target molecule by aptamer-based target cell sorting and analysis of clinical samples; and identifying the aptamer's target molecule using MS and aptamer-based target enrichment. This stratagem not only shortens the time for the clinical application of aptamers but also enables a more targeted and efficient discovery of biomarkers. [ABSTRACT FROM AUTHOR]

A novel molecularly imprinted polymer (MIP) based on hydrophobic CdSe quantum dots (QDs) was synthesized using a one-pot room-temperature reverse microemulsion polymerization, and this polymer was applied as a molecular recognition element to construct a ractopamine (RAC) optosensor. Here, hydrophobic CdSe QDs were first introduced to the hydrophilic analyte-imprinted polymer for highly selective and sensitive detection of RAC via the change in fluorescence intensity, because of the high-quality hydrophobic QDs with high quantum yield, sharp photoluminescence spectra and chemical and fluorescent stability. Under optimal conditions, the relative fluorescence intensity of MIP based on hydrophobic QDs decreased linearly with the increasing concentration of RAC in the range of 1.21×10−9–3.03×10−6 molL−1 with a detection limit of 7.57×10−10 molL−1, and the precision for five replicate detections of 1.51×10−8 molL−1 RAC was 2.09% (relative standard deviation). The proposed method was successfully applied for the determination of trace RAC in pork samples, with good recoveries ranging from 82.79% to 97.23%. [ABSTRACT FROM AUTHOR]

Micro/nanomaterials display considerable potential for increasing the sensitivity of lateral flow immunoassay (LFIA) by acting as 3D carriers for both antibodies and signals. The key to achieving high detection sensitivity depends on the probe's orientation on the material surface and its multivalent biomolecular interactions with targets. Here, we engineer Lactococcus lactis as the bacterial microcarrier (BMC) for a multivalent immunorecognition probe that was genetically programmed to display multifunctional components including a phage-screened single-chain variable fragment (scFv), an enhanced green fluorescent protein (eGFP), and a C-terminal peptidoglycan-binding domain (AcmA) anchored on BMC through the cell wall peptidoglycan. The innovative design of this biocarrier system, which incorporates a lab-on-a-chip microfluidic device, allows for the rapid and non-destructive self-assembly of the multivalent scFv-eGFP-AcmA@BMC probe, in which the 3D structure of BMC with a large peptidoglycan surface area facilitates the precisely orientated attachment and immobilization of scFv-eGFP-AcmA. This leads to a remarkable fluorescence aggregation amplification effect in LFIA, outperforming a monovalent 2D scFv-eGFP-AcmA probe for florfenicol detection. By designing a portable sensing device, we achieved an exceptionally low detection limit of 0.28 pg/mL and 0.21 pg/mL for florfenicol in lake water and milk sample, respectively. The successful microfabrication of this biocarrier holds potential to inspire innovative biohybrid designs for environment and food safety biosensing applications. [ABSTRACT FROM AUTHOR]

For the first time, a quantitative photothermal-sensing immunochromatographic sensor (PT-ICS) is described using Au nanoparticle-enhanced two-dimensional black phosphorus (BP-Au) as signal component for the photothermal-sensing antibody probe. BP-Au has good photothermal properties at 808 nm, and the photothermal conversion efficiency of the BP-Au nanosheet increased by 12.9% over the black phosphorus nanosheet alone. In addition, the antibody was more easily coupled to this nanosheet due to the good physical adsorption capacity of Au nanoparticles. We used this PT-ICS to detect veterinary antibiotics enrofloxacin (ENR), the photothermal-sensing antibody probe was competitive captured by ENR target and antigen coating on test (T) lines of the sensor. This process was exothermic under an 808 nm laser, and the thermal energy decreased as the ENR in the sample increased. This thermal energy was recorded by an infrared thermal imager or an infrared thermometer, and the concentration of the ENR residues in animal-derived foods was obtained by analyzing the temperature changes in T-lines. Under optimal conditions, the PT-ICS exhibited sensitive and specific detection of ENR from 0.03 μg/L to 10 μg/L with detection limits of 0.023 μg/L. The results agreed well with a commercial enzyme-linked-immunosorbent assay kit. This PT-ICS provided a promising strategy for the detection of ENR residues in animal-derived foods and expected to be used for the detection of other highly sensitive biomacromolecules. fx1 • Au nanoparticle-enhanced black phosphorus nanosheet was synthesized and used as photothermal-sensing immuno-probe. • Developed a novel photothermal sensing immunochromatographic sensor for ENR detecting in animal derived food samples. • Quantitative analysis was realized by an infrared thermometer or a thermal imager. • The photothermal-sensing immunochromatographic sensor increases the sensitivity and improves the detection range of an assay. [ABSTRACT FROM AUTHOR]

Abstract For the determination of dopamine (DA) in serum samples, a quenching-type electrochemiluminescence sensor (MIECLS) was constructed in this study. Upconversion nanoparticles (UCNPs) enhanced by covalent organic frameworks (COFs)-based hybrid and oligoaniline-crosslinked gold nanoparticles (AuNPs) imprinting recognition sites were introduced in electrochemiluminescence (ECL) system at the first time. The porous COFs-based hybrid with large specific surface area was modified on the electrode firstly to hold more UCNPs and imprinting recognition sites. AuNPs was employed in the developed sensor for two objectives: 1) AuNPs on the COFs-based hybrid enabled the hybrid to tunnel the electrons, which helped to improve the ECL intensity; 2) AuNPs-based thioaniline units (PATP@AuNPs) electropolymerized on the electrode in the presence of template to form oligoaniline-crosslinked AuNPs matrix. Then the exclusion of template from matrix yielded the molecularly three-dimensional imprinted contours with conductivity, which facilitated specific recognition and further amplified the ECL. The double recognition mode in this work involves the recognition effect of imprinted contours and quenching effect of o -benzoquinone species. The quantum chemical calculation was performed to analyze the possible recognition and the binding mechanisms of molecularly three-dimensional imprinted contours. The results showed imprinted contours could bind the targets by complementary spatial cavities and weak interactions. The proposed approach yielded a wide detection range (10–14–10−6 M), low limit of detection (LOD = 2 ×10–15 M) and acceptable recoveries (93.25–112.97%) in rat serum sample, demonstrating that the developed method holds great promise to be applied to DA detection in practical samples. Highlights • CTpBD-Au enhanced UCNPs and oligoaniline-crosslinked AuNPs imprinting recognition sites were introduced in ECL system. • The double recognition was constructed by the recognition effect of imprinted contours and quenching effect for ECL. • The proposed approach yields a wide detection range, low limit of detection and acceptable recoveries in rat serum sample. [ABSTRACT FROM AUTHOR]

A molecularly imprinted electrochemiluminescence sensor (MIECLS) is constructed to selectively detect clenbuterol (CLB) based on boron nitride quantum dots@gold nanoflowers/silver nanowires (BNQDs@AuNFs/AgNWs). The abundant amino and hydroxyl groups on the surface of the BNQDs generate an electrostatic self-assembly effect with the multi-tipped spatial structure of AuNFs, constituting a novel nanoscale co-reaction accelerator (NCRA) with high activity and large load capacity. An NCRA embedded in the network structure of the AgNW luminophores significantly promotes the reduction of peroxydisulfate (S 2 O 8 2−) to sulfate anion radicals (SO 4 −•) through the catalysis of amino groups and boron radicals (B•) and the electron acceleration of AuNFs while also reducing the reaction distance between SO 4 −• and AgNWs−•, realizing the multiple synergistic amplification of the electrochemiluminescence (ECL) signal. Imprinted cavities in the molecularly imprinted polymers (MIPs) prepared by electropolymerization can generate a "blocking-effect" by recognizing CLB, realizing ECL signal quenching. Analytical results indicate that the established MIECLS detects CLB in a line concentration range of 0.5–50000 nM and detection limit of 0.00693 nM. The spiked recoveries are 85.90%–97.77%, with the relative standard deviations (RSD) under 5.1%, consistent with those of high-performance liquid chromatography (HPLC). This work demonstrates that an efficient NCRA can significantly enhance the output of the ECL signal in collaboration with the original luminophore, providing a new method to realize the ultra-detection of targeted substances by MIECLS. [Display omitted] • AgNWs with excellent conductivity and transparency were used as ECL luminophores. • Green BNQDs can be electrostatically assembled onto surface of multi-tipped AuNFs. • Novel NCRA based on BNQDs@AuNFs had multiply synergistic catalyses of S 2 O 8 2−. • The MIIECLS showed selective "blocking-effect" detection strategy for clenbuterol. • The sensitive MIECLS had been successfully applied to the detection of real samples. [ABSTRACT FROM AUTHOR]

A simple, efficient and sensitive molecularly imprinted electrochemiluminescence sensor (MIECLS) based on reduced graphene oxide (rGO) and upconversion nanoparticles (UCNPs) was developed for determination of clenbuterol (CLB). In this study, rGO generated by electrodeposition of graphene oxide not only acted as carrier for immobilizing UCNPs, but also had a significant impact in boosting electrochemiluminescence (ECL) response of UCNPs thanks to its high conductivity, superior electron transport rate and large specific surface area. UCNPs as an advanced ECL emitter possessed wonderful ECL performance. Furthermore, the introduction of molecularly imprinted polymers (MIP) endowed the ECL sensor a new character of specifically identifying analyte CLB. Under the optimal experimental conditions, the ECL signal was proportional to the logarithm of CLB concentration in the range of 10 nM to 100 µM with a low detection limit of 6.3 nM. The proposed MIECLS combining the advantages of UCNPs-ECL and MIP exhibited good sensitivity, desirable selectivity and favorable stability, indicating enormous potential in the future of food safety detection. [ABSTRACT FROM AUTHOR]

Since HCV and HIV share a common transmission path, high sensitive detection of HIV and HCV gene is of significant importance to improve diagnosis accuracy and cure rate at early stage for HIV virus-infected patients. In our investigation, a novel nanozyme-based bio-barcode fluorescence amplified assay is successfully developed for simultaneous detection of HIV and HCV DNAs with excellent sensitivity in an enzyme-free and label-free condition. Here, bimetallic nanoparticles, PtAu NPs , present outstanding peroxidase-like activity and act as barcode to catalyze oxidation of nonfluorescent substrate of amplex red (AR) into fluorescent resorufin generating stable and sensitive "Turn On" fluorescent output signal, which is for the first time to be integrated with bio-barcode strategy for fluorescence detection DNA. Furthermore, the provided strategy presents excellent specificity and can distinguish single-base mismatched mutant from target DNA. What interesting is that cascaded INHIBIT-OR logic gate is integrated with biosensors for the first time to distinguish individual target DNA from each other under logic function control, which presents great application in development of rapid and intelligent detection. [ABSTRACT FROM AUTHOR]

A novel molecular imprinting fluorescent sensor synthesized avoiding complicated design and synthesis of a fluorescent functional monomer, and without the extra cost of additional fluorescence materials (quantum dots, carbon dots and fluorescein), only by a click reaction making the common protein imprinting polymers upgraded to a fluorescent sensing material which can be applied to the fast detection of glycoproteins. In this study, a common protein imprinted polymer (protein-IP) which containing a disulfide bond in the imprinted cavities was first synthesized, then the disulfide linkage was cleaved by reduction, the exposed thiol groups were used for the click reaction to introduce 4-vinyl phenylboronic acid to improve the imprinted recognition for glycoproteins and convert recognition behavior into fluorescent signal simultaneously in one-step site-specific post-imprinting modifications (PIMs). Good linear relationship was obtained in the range of 0.1–10 mg mL −1 by fluorescence assay, comparison of the adsorption capacity of the MIPs before and after modification proved that the method has no negative impact on the imprinting effect and exhibit good imprinting effect for glycoproteins of different molecular size and weight. [ABSTRACT FROM AUTHOR]

Sensitive and rapid detection of Escherichia coli O157:H7, one of the most notorious bacterial pathogens, is urgently needed for public health protection. Yet, the existing methods are either lack of speed or limited in discriminating viable and dead cells. Using a recombinant bacteriophage, here we report the development of a rapid and sensitive method for live E. coli O157:H7 detection. First, the wild-type PP01 phage was engineered with a tetracysteine (TC)-tag fused with the small outer capsid (SOC) protein. Then, this PP01-TC phage was used to inoculate bacterial sample for 30 min. Specific infection and rapid replication of PP01-TC phage in viable E. coli O157:H7 host cell yields a large number of progeny phages with capsids displaying TC tags that can be fluorescently labeled by a membrane permeable biarsenical dye (FlAsH). The bright green fluorescence of single E. coli O157:H7 cells can be readily detected by flow cytometry (FCM) and fluorescence microscopy. High specificity of the assay was verified with seven other bacterial strains. Practical application in E. coli O157:H7 detection in drinks was successfully demonstrated with artificially contaminated 100% apple juice. In less than three hours (including sample preconcentration) and with 40 mL of sample volume, as low as 1 cfu/mL E. coli O157:H7 can be detected in the presence of large excess of other nontarget bacteria via fluorescence microscopic measurement. The as-developed TC-PP01-FlAsH approach shows a great potential in the safeguard of liquid food products by providing rapid, sensitive, and specific detection of live E. coli O157:H7. [ABSTRACT FROM AUTHOR]

In this study, the three nanomaterials: colloidal gold, nanogold-polyaniline-nanogold microspheres (GPGs) and colloidal carbon were respectively labeled with the antibody against salbutamol (SAL). We aimed to develop immunochromatographic strips with these nanomaterial labels and determine their performance in visual detection of SAL. For the colloidal gold-based strip, the detection limit of SAL was 1.0 µg L −1 in standard solution and 5.0 µg kg −1 in meat samples. For the GPG- and colloidal carbon-based strips, the limit of detection was 2.0 µg L −1 in standard solution and 10 µg kg −1 in meat samples. The results obtained using the test strips were found to be highly consistent with those obtained using a commercial kit, indicating the high accuracy of these strips. The three strips were also found to be stable up to 18 weeks under laboratory conditions. In terms of sensitivity, the colloidal gold-based strip was slightly better than the other two. For the GPG- and colloidal carbon-based strips, the difference between the results obtained for different batches was small (high consistency), and the stability was much better than that of the colloidal gold-based one. Our results indicate that colloidal carbon can be used as a label in immunochromatographic tests; it can also help reduce the cost involved and scale-up the production. The use of immunochromatographic test strips labeled with colloidal carbon can be a rapid and inexpensive method for SAL assays in on-site applications. [ABSTRACT FROM AUTHOR]

A novel strategy was developed to prepare double responsive smart upconversion fluorescence material for highly specific enrichment and sensing of glycoprotein. The novel double responsive smart sensing material was synthesized by choosing Horse radish peroxidase (HRP) as modal protein, the grapheme oxide (GO) as support material, upconversion nanoparticles (UCNPs) as fluorescence signal reporter, N-isopropyl acrylamide (NIPAAM) and 4-vinylphenylboronic acid (VPBA) as functional monomers. The structure and component of smart sensing material was investigated by transmission electron microscopy (TEM), Scanning electron microscopy (SEM), X-ray photoelectron spectroscopic (XPS) and Fourier transform infrared (FTIR), respectively. These results illustrated the smart sensing material was prepared successfully. The recognition characterizations of smart sensing material were evaluated, and results showed that the fluorescence intensity of smart sensing material was reduced gradually, as the concentration of protein increased, and the smart sensing material showed selective recognition for HRP among other proteins. Furthermore, the recognition ability of the smart sensing material for glycoprotein was regulated by controlling the pH value and temperature. Therefore, this strategy opens up new way to construct smart material for detection of glycoprotein. [ABSTRACT FROM AUTHOR]

A novel approach to synthesize molecularly imprinted polymer (MIP) nanoparticles using a MIL-101 support (a type of metal–organic framework) is reported herein for the first time; the sample is referred as MIL@MIP. The nanoparticles were well distributed within the polymer film, and exhibit an octahedral shape, satisfied thermal stability, and a high specific surface area (SSA) of 1579.43 m 2 g −1 . The adsorption behavior of MIL@MIP toward metolcarb in aqueous solution was subsequently examined. The synthesized MIL@MIP displayed satisfactory high transfer mass rates and a high selective adsorption affinity for metolcarb. Based on these results, a quartz crystal microbalance (QCM) sensor based on MIL@MIP was subsequently constructed and examined for the sensitive detection of metolcarb. Under optimal conditions, the detection limit of the system assessed in pear juice was 0.0689 mg L −1 within a linear concentration range of 0.1–0.9 mg L −1 . MIL@MIP–QCM system combines the advantages of MIL-101 and molecularly imprinted technology (MIT), thereby achieving high detection sensitivity and selectivity. The current findings suggest the potential of MIL@MIP for detecting trace level pesticides and veterinary drugs for food safety and environmental control. [ABSTRACT FROM AUTHOR]

A novel fluorescence material with thermo-sensitive for the enrichment and sensing of protein was successfully prepared by combining molecular imprinting technology with upconversion nanoparticles (UCNPs) and metal-organic frameworks (MOFs). Herein, the UCNPs acted as signal reporter for composite materials because of its excellent fluorescence property and chemical stability. MOFs were introduced to molecularly imprinted polymer (MIP) due to its high specific surface area which increases the rate of mass transfer relative to that of traditional bulk MIP. The thermo-sensitive imprinted material which allows for swelling and shrinking with response to temperature changes was prepared by choosing Bovine hemoglobin (BHB) as the template, N-isopropyl acrylamide (NIPAAM) as the temperature-sensitive functional monomer and N,N-methylenebisacrylamide (MBA) as the cross-linker. The recognition characterizations of imprinted material-coated UCNPs/MOFs (UCNPs/MOFs/MIP) were evaluated, and the results showed that the fluorescence intensity of UCNPs/MOFs/MIP reduced gradually with the increase of BHB concentration. The fluorescence material was response to the temperature. The adsorption capacity was as much as 167.6 mg/g at 28 °C and 101.2 mg/g at 44 °C, which was higher than that of traditional MIP. Therefore, this new fluorescence material for enrichment and sensing protein is very promising for future applications. [ABSTRACT FROM AUTHOR]

A novel sensitive fluorescent sensor for determination of sterigmatocystin (ST), which was based on carbon dots-embedded molecularly imprinted polymer (CDs@MIP), was prepared by an efficient one-pot reaction. First, highly blue luminescent CDs were synthesized via a one-step reaction. Then, through a non-hydrolytic sol–gel process, MIP was formed on the CDs surface in the presence of 1,8-dihydroxyanthraquinone as an alternative template molecule to obtain CDs@MIP. The CDs acted as antennas for signal amplification and optical readout, and the MIP coated on the CDs surface provided specific binding sites for ST. The performance of CDs@MIP was compared with that of CDs embedded in non-imprinted polymer (CDs@NIP). CDs@MIP exhibited high selectivity and sensitivity toward ST. Under optimized conditions, the relative fluorescence intensity of CDs@MIP decreased linearly with the concentration of ST from 0.05 to 2.0 mg L −1 with a detection limit of 0.019 mg L −1 ( S / N =3) and the precision for five replicate detections of 0.10 mg L −1 ST was 2.31%. The sensor was also used to determine the content of ST in grains with satisfactory results. [ABSTRACT FROM AUTHOR]

A novel strategy is reported for the fabrication of bis-aniline-crosslinked Au nanoparticles (NPs)-CdSe/ZnS quantum dots (QDs) array composite by facil one-step co-electropolymerization of thioaniline-functionalized AuNPs and thioaniline-functionalized CdSe/ZnS QDs onto thioaniline-functionalized Au elctrodes (AuE). Stable and enhanced cathodic electrochemiluminescence (ECL) of CdSe/ZnS QDs is observed on the modified electrode in neutral solution, suggesting promising applications in ECL sensing. An advanced ECL sensor is explored for detection of 2-methyl-4-chlorophenoxyacetic acid (MCPA) which quenches the ECL signal through electron-transfer pathway. The sensitive determination of MCPA with limit of detection (LOD) of 2.2 nmol L −1 (S/N=3) is achieved by π-donor–acceptor interactions between MCPA and the bis-aniline bridging units. Impressively, the imprinting of molecular recognition sites into the bis-aniline-crosslinked AuNPs–CdSe/ZnS QDs array yields a functionalized electrode with an extremely sensitive response to MCPA in a linear range of 10 pmol L −1 –50 μmol L −1 with a LOD of 4.3 pmol L −1 ( S/N=3). The proposed ECL sensor with high sensitivity, good selectivity, reproducibility and stability has been successfully applied for the determination of MCPA in real samples with satisfactory recoveries. In this study, ECL sensor combined the merits of QDs-ECL and molecularly imprinting technology is reported for the first time. The developed ECL sensor holds great promise for the fabrication of QDs-based ECL sensors with improved sensitivity and furthermore opens the door to wide applications of QDs-based ECL in food safety and environmental monitoring. [ABSTRACT FROM AUTHOR]

A triple-dimensional sensing chip is developed based on simultaneous utilization of fluorescence (FL), electrochemical (ECL) and mass-sensitivity (MS) properties of a novel nanocomposites. The sensing nanomaterial is composed of CdSe/ZnS quantum dots (QDs) and graphene through a one-pot room-temperature reverse microemulsion polymerization. Here, full integration of QDs and graphene on one chip provides triple-dimensional sensing signals. It enables quick and accurate discrimination of eight analytes in a “lab-on-a-nanomaterial” approach and notably improves the overall sensor performance. Unknown samples randomly taken from the training set at concentrations of 0.7 μM are successfully classified by principal component analysis (PCA) with accuracies of 92.5%, compared with the high performance liquid chromatography (HPLC) method. We further apply it to discriminate eight antioxidants from real oil samples, and explore the mechanism. [ABSTRACT FROM AUTHOR]

A novel method combined the high selectivity of molecular imprinting technology with the strong fluorescence property of upconversion nanoparticles (UCNPs) for sensing of Cytochrome c (Cyt c) was proposed. The molecularly imprinted material-coated upconversion nanoparticles (UCNPs@MIP) were obtained by in situ coating Cyt c imprinted materials to the surface of the carboxyl modified UCNPs through sol–gel technique. The structure and component of the prepared UCNPs@MIP was investigated by transmission electron microscopy (TEM), power X-ray diffraction (XRD), energy-dispersive X-ray analysis (EDXA) and X-ray photoelectron spectroscopic (XPS). The TEM showed the diameter of UCNPs was 40 nm, and thickness of MIP was 5–10 nm. The fluorescence intensity of UCNPs@MIP reduced gradually with the increase of Cyt c concentration. Under optimum conditions, the imprinting factor is 3.19, and the UCNPs@MIP showed selective recognition for Cyt c among other proteins such as bovine serum albumin (BSA) and Lysozyme (Lyz). Therefore, this new method for sensing protein is very promising for future applications. [ABSTRACT FROM AUTHOR]

A molecularly imprinted quartz crystal microbalance (QCM) sensor for ractopamine (RAC) detection was developed by electrodepositing a poly-o-aminothiophenol membrane on an Au electrode surface modified by self-assembled Au nanoparticles (AuNPs). The modified electrodes were characterized by cyclic voltammetry, electrochemical impedance spectroscopy and scanning electron microscopy. This molecularly imprinted QCM sensor showed good frequency response in RAC binding measurements and the introduction of AuNPs demonstrated performance improvements. Frequency shifts were found to be proportional to concentration of RAC in the range of 2.5×10−6 to 1.5×10−4 molL−1 with a detection limit of 1.17×10−6 molL−1 (S/N=3). The sensor showed a good selective affinity for RAC (selectivity coefficient >3) compared with similar molecules and good reproducibility and long-term stability. This research has combined the advantages of high specific surface area of AuNPs, high selectivity from molecularly imprinted electrodeposited membrane and high sensitivity from quartz crystal microgravimetry. In addition, the modified electrode sensor was successfully applied to determine RAC residues in spiked swine feed samples with satisfactory recoveries ranging from 87.7 to 95.2%. [ABSTRACT FROM AUTHOR]

Abstract: Single-cell analysis is vital in providing insights into the heterogeneity in molecular content and phenotypic characteristics of complex or clonal cell populations. As many essential proteins and most transcription factors are produced at a low copy number, analytical tools with superior sensitivity to enable the analysis of low abundance proteins in single cells are in high demand. β-galactosidase (β-gal) has been the standard cellular reporter for gene expression in both prokaryotic and eukaryotic cells. Here we report the development of a high-throughput method for the single-cell analysis of low copy number β-gal proteins using a laboratory-built high-sensitivity flow cytometer (HSFCM). Upon fluorescence staining with a fluorogenic substrate, quantitative measurements of the basal and near-basal expression of β-gal in single Escherichia coli BL21(DE3) cells were demonstrated. Statistical distribution can be determined quickly by analyzing thousands of individual cells in 1–2min, which reveals the heterogeneous expression pattern that is otherwise masked by the ensemble analysis. Combined with the quantitative fluorometric assay and the rapid bacterial enumeration by HSFCM, the β-gal expression distribution profile could be converted from arbitrary fluorescence units to protein copy numbers per cell. The sensitivity and speed of the HSFCM offers great capability in quantitative analysis of low abundance proteins in single cells, which would help gaining a deeper insight into the heterogeneity and fundamental biological processes in microbial populations. [Copyright &y& Elsevier]

Quinoxaline-2-carboxylic acid (QCA) is difficult to measure since only trace levels are present in commercial meat products. In this study, a rapid, sensitive and selective molecularly imprinted electrochemical sensor for QCA determination was successfully constructed by combination of a novel modified glassy carbon electrode (GCE) and differential pulse voltammetry (DPV). The GCE was fabricated via stepwise modification of multi-walled carbon nanotubes (MWNTs)-chitosan (CS) functional composite and a sol–gel molecularly imprinted polymer (MIP) film on the surface. MWNTs-CS composite was used to enhance the electron transfer rate and expand electrode surface area, and consequently amplify QCA reduction electrochemical response. The imprinted mechanism and experimental parameters affecting the performance of MIP film were discussed in detail. The resulting MIP/sol–gel/MWNTs-CS/GCE was characterized using various electrochemical methods involving cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and DPV. The sensor using MIP/sol–gel/MWNTs-CS/GCE as working electrode showed a linear current response to the target QCA concentration in the wide range from 2.0×10−6 to 1.0×10−3 molL−1 with a low detection limit of 4.4×10−7 molL−1 (S/N=3). The established sensor with excellent reproductivity and stability was applied to evaluate commercial pork products. At five concentration levels, the recoveries and standard deviations were calculated as 93.5–98.6% and 1.7–3.3%, respectively, suggesting the proposed sensor is promising for the accurate quantification of QCA at trace levels in meat samples. [ABSTRACT FROM AUTHOR]

This work prepared a novel ionic liquid stabilized molecularly imprinted optosensing material based on quantum dots (QDs) and graphene oxide (GO) composites using a one-step polymerization for highly selective and sensitive specific recognition of vitamin E (VE). Here, GO was first introduced to the molecularly imprinted polymer (MIP) because of its ultra-high specific surface area which increases the rate of mass-transfer relative to that of traditional bulk MIP. The ionic liquid was used to provide the desired surface binding groups between the QDs and the GO, but also to improve their fluorescence stability by virtue of its high thermal and chemical stability. Under optimal conditions, the relative fluorescence intensity of MIP decreased linearly with the increasing concentration of VE in the range of 2.30×10−2–9.20×102 μM with a detection limit of 3.5nM and the precision for five replicate detections of 92μM VE was 1.67% (relative standard deviation). At three concentration levels, the recoveries for two samples achieved were 93.00–100.14% and 92.00–102.00%, respectively. [ABSTRACT FROM AUTHOR]

A hydrophobic carboxyl functionalized phosphonium-based ionic liquid (IL) ((5 -carboxypentyl) triphenylphosphonium bis (trifluoromethyl)sulfonyl) amide, TPP-HA[TFSI]) was synthesized through a simple hydrothermal approach. Based on the π-π and cation-π interactions with multi-wall carbon nanotubes (MWCNTs), a TPP-HA[TFSI]@MWCNTs hybrid was prepared to immobilize hemoglobin (Hb) to fabricate a simple and effective electrochemical sensing platform for the detection of methyl parathion (MP) in vegetables. Spectroscopic and electrochemical results show that TPP-HA[TFSI]@MWCNTs substrate synergistically provided a good biocompatible microenvironment for Hb, and the hydrophobicity of TPP-HA[TFSI] and the π-π interaction and hydrogen bonding between TPP-HA[TFSI]@MWCNTs, Hb and nafion (NF) were conducive to maintain the stability and integrity of the modified electrode interface. The TPP-HA[TFSI]@MWCNTs with large surface area and high conductivity promoted the exposure of the electroactive center of Hb and the direct electron transfer between Hb and the electrode, which effectively amplified the electrochemical signal and improved the sensitivity of MP detection. The constructed electrochemical sensing platform had a wider linear range (2–14 ng mL-1) and a lower detection limit (0.62 ng mL-1) for MP, and had acceptable repeatability, reproducibility, stability and anti-interference ability. This results indicated that the phosphonium-based ILs functionalized MWCNTs was an effective substrate for the immobilization of biological components, which have broad prospect in the construction of electrochemical sensing interfaces. • The hydrophobic phosphonium-based ionic liquid (TPP-HA[TFSI]) was synthesized through a simple hydrothermal approach. • The TPP-HA[TFSI]@MWCNTs hybrid was prepared based on π-π and cation-π interactions between MWCNTs and TPP-HA[TFSI]. • The TPP-HA[TFSI]@MWCNTs hybrid has good immobilization ability and improved catalytic effect on hemoglobin (Hb). • An electrochemical sensing platform with enhanced stability and conductivity was applied to develop. • The NF/Hb/TPP-HA[TFSI]@MWCNTs/GCE is simple to prepare and exhibits a good performance in methyl parathion detection. [ABSTRACT FROM AUTHOR]

Abstract: This study describes the development of a sensor with molecularly imprinted polymer (MIP) sensitized with MWCNTs and Salen-Co(III) as recognition element for methimazole (MMI) determination. This is the first report of MWCNTs and Salen-Co(III) in MIP being used to enhance its conductivity and catalytic activity in the electrochemical oxidation process. The electrocatalytic mechanism of MMI was explained in detail by evaluating the obtained voltammograms at various potential sweep rates and pH of buffer solutions. A stable, sensitive analytical method of differential pulse voltammetry (DPV) was developed using the prepared MWCNTs–Salen-Co(III)–MIP electrode for the determination of amounts of MMI, resulting in a linear range of 0.5–6.0mgL−1 with detection limit of 0.048mgL−1. It was also successfully applied for MMI determination in tablets and spiked urine sample. At three concentration levels, the recoveries for two samples were achieved to 94.0–100.1% and 87.8–101.8%, respectively. Analytical reproducibility and stability of the developed method were also evaluated by RSD, which were calculated as 4.6–6.6% and 2.0–6.1% (n =3), respectively. [Copyright &y& Elsevier]

Cell-free synthetic biology provides a promising strategy for developing high-performance biosensors by integrating with advanced testing technologies. However, the combination of synthetic biology with electrochemical testing techniques is still underdeveloped. Here, we proposed an electrochemical biosensor for the label-free and ultrasensitive detection of target protease biomarker by coupling a protease-responsive RNA polymerase (PR) for signal amplification. Taking tumor biomarker matrix metalloprotease-2 (MMP-2) as a model protease, we employed PR to transduce each proteolysis reaction mediated by MMP-2 into multiple programmable RNA outputs that can be captured by the DNA probes immobilized on a gold electrode. Moreover, the captured RNAs are designed to contain a guanine-rich sequence that can form G-quadruplex and bind to hemin in the presence of potassium ions. In this scenario, the activity of MMP-2 is converted and amplified into the electrochemical signals of hemin. Under the optimal conditions, this PR-based electrochemical biosensor enabled the sensitive detection of MMP-2 in a wide linear dynamic range from 10 fM to 1.0 nM, with a limit of detection of 7.1 fM. Moreover, the proposed biosensor was further applied in evaluating MMP-2 activities in different cell cultures and human tissue samples, demonstrating its potential in the analysis of protease biomarkers in complex clinical samples. • Integrating protease-triggered transcription to fabricate an electrochemical sensor. • Amplified output of RNA G-quadruplexes enabled label-free electrochemical detection. • Detection of matrix metalloproteinase-2 with femtomolar sensitivity was achieved. • Specific analysis of matrix metalloproteinase-2 in human tissue samples was performed. [ABSTRACT FROM AUTHOR]

Bacterial infection is a growing public health concern and causes a huge medical and financial burden. It is of significance to efficiently construct multifunctional platforms for bacterial point-of-care testing (POCT) and elimination. Herein, near-infrared (NIR) light-responded vancomycin-doped prussian blue nanoparticles (PB-VAN NPs) with high efficient photothermal conversion was synthesized for binding, dual-mode portable detection, and elimination of bacteria. The PB-VAN NPs can bind to the surface of Gram-positive bacteria such as Staphylococcus aureus (S. aureus), forming complex of PB-VAN NPs / S. aureus. After being centrifugated, the suspension solution of PB-VAN NPs can stimulate perfluorohexane (PFH) to rapidly release oxygen (O 2) under NIR irradiation. Thus, the bacteria can be sensitively detected with portable pressure meter as signal reader, reporting a limit of detection (LOD) of 1.0 CFU mL-1. On the other side, the sediment of PB-VAN NPs / S. aureus can be detected via thermal camera, reporting a LOD of 1.0 CFU mL-1. Interestingly, the bacteria can be effectively inactivated with the local temperature elevation during temperature-based detection. The antibacterial efficiency reaches as high as 99.8%. The developed multifunctional nanoplatform not only provides a straightforward "mix-then-test" way for portable detection of bacteria with high sensitivity, also realizes high efficiency elimination of bacteria simultaneously. The developed strategy was further applied for promoting wound healing of bacteria-infected mice. • A multifunctional nanoplatform was constructed for POCT and elimination of bacteria. • The POCT was implemented either with portable pressure meter or thermal camera. • Bacteria can be high efficiently eliminated via NIR photothermal treatment. • The developed strategy can promote wound healing of bacteria-infected mice. [ABSTRACT FROM AUTHOR]

With increasing concerns related to the abuse of antibiotics in livestock production worldwide, simple and rapid screening methods for monitoring antibiotics in animal-derived foods are highly desirable. In this study, we propose a facile synthesis strategy for gold nanoclusters (AuNCs) exhibiting remarkable optical properties by employing ovalbumin (OVA) as the template. The OVA-stabilized AuNCs (AuNCs@OVA) manifest intriguing multicolour fluorescence and a gradually declining fluorescence intensity at 650 nm with an increasing concentration of tetracycline family antibiotics (TCs) including tetracycline, chlorotetracycline, oxytetracycline, and doxycycline, which are a widely used class of antibiotics for treating infections in food-producing animals. This performance makes AuNCs@OVA particularly attractive as a broad-spectrum detector for TCs sensing, and we demonstrate that this simple sensing procedure can be realized in real time by directly mixing the target sample and AuNCs@OVA components. Based on this sensing strategy, a microfluidic lab-on-a-chip platform was constructed for the ultrarapid detection of TCs within 30 s. The detection limit was determined to be 0.09 μg/mL in chicken muscle extract, with the recovery ranging from 86.20% to 93.57% in spiked samples. This work provides not only a broad-spectrum sensing strategy for TCs but also a pump-free microfluidic chip with the advantages of being portable, ultrarapid, and low cost, offering a viable alternative for on-the-spot ultrarapid screening of TCs. • A facile synthesis strategy for ovalbumin-stabilized gold nanocluster (AuNCs@OVA). • A novel real-time sensing strategy for TCs using AuNCs@OVA as a broad-spectrum detector. • A single small lab-on-a-chip device for easy and ultrarapid detection of TCs. [ABSTRACT FROM AUTHOR]