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A solvent-assisted ESIPT fluorescent dye was synthesized and used as a probe (2-PPN) for the detection of F−/Ag+ and high-resolution imaging of the cilia in live cells. The developed ESIPT fluorophore exhibited strong tautomeric fluorescence in protic solvents and normal emission in aprotic solvents, which is a significant departure from that of conventional intramolecular ESIPT compounds. The H-binding interaction of F− and the chelation of Ag+ with the ESIPT module of 2-PPN resulted in significant tautomeric emission quenching. From this basis, the 2-PPN-based assays for the detection of F− and Ag+ were established. The detection limit for F− and Ag+ sensing is 2.4 nM and 1.5 nM, respectively. The selective experimental results showed that no tautomeric fluorescence change of 2-PPN could be observed in the presence of the other inorganic ions in the same medium, revealing high selectivity of 2-PPN to F− and Ag+. Furthermore, MTT assay experiments proved that the probe 2-PPN exhibited low cytotoxicity and good cell membrane permeability. The probe was also further successfully utilized to image the cilia in vitro MCF7 cells, displaying its high-resolution imaging performance. Graphical abstract

Strongly green fluorescent carbon dots (CQDs) have been directly synthesized from 2,4-diaminophenylhydrazine and 2-hydroxy-5-methylisophthalaldehyde through a facile solvothermal method. The novel CQDs exhibit high fluorescence quantum yield and excellent water solubility due to the abundant amino and hydroxy groups on their surface. The use of the as-prepared CQDs combined with Cu2+ constructed a “turn-on” switch cysteine-responsive nanoprobe. In the CQDs-Cu2+ assemblies, the binding of Cu2+ to CQDs results in the fluorescence quenching of CQDs by electron transfer mechanism, while the addition of cysteine leads to the fluorescence recovery because of the competitive binding between cysteine and CQDs to Cu2+. The nanoprobes showed high sensitivity to cysteine with the detection limit of 2.6 nmol L−1. The selectivity investigation results demonstrated that the Cu2+-integrated nanoparticles were highly selective toward cysteine over the other amino acids and biologically related metal ions. The proposed nanoprobe was then employed for detecting the recovery of cysteine in rabbit serum and plasma samples and imaging the cysteine in cancer cells, and the recovery was found to be 98.2–104.0%. This “synthesis-modification integration” strategy for the fabrication of CQDs may offer a new sight for the preparation of multifunctional nanostructures and broadening the application of CQDs in bioimaging.

As a new member of the carbon quantum-dot family, fluorescent nitrogen-rich quantum dots (NRQDs) were prepared by a mixed solvothermal method using 2-azidoimidazole and aqueous ammonia as reactants. These NRQDs are rich in nitrogen up to 40.2%, which are endowed with high fluorescence quantum yield, good photostability, water-solubility and favourable biocompatibility. We further explored the use of NRQDs combined with Cu2+ as a nanoprobe for sensing fluorescently of cysteine (Cys) in complex biological samples. In this sensing system, the fluorescence is significantly quenched via energy transfer from NRQDs to Cu2+ for the coordination of amino-containing groups with Cu2+. The strong affinity between Cu 2+ and Cys leads to the formation of Cu2+-Cys complexes and cause the detachment of Cu2+ from the surface of NRQDs, thus the fluorescence of NRQDs recover. This nanoprobe allows analysis of Cys by modulating the switch of the fluorescence of NRQDs with a detection limit of 5.3nM. As expected, the proposed NRQDs-Cu2+complex-based nanoprobes were successfully applied for the determination of Cys in human serum and plasma samples with recoveries ranging from 97.2% to 105.7%. The probe ensemble was also successfully applied to imaging of Cys in living cells with satisfactory results, which shows strong potential for clinical diagnosis.

A natural product, stilbene glycoside (2,3,5,4'-tet rahydroxy- diphenylethylene-2-O-glucoside, TBG), has been evaluated for the first time as a potential substrate for horseradish peroxidase (HRP )-catalyzed fluorogenic reactions. The properties of TBG as a fluorogenic substrate fo r HRP and its application in a fluorometric enzyme-linked immunosensing system were compared with commercially available substrates such as p-hydroxyphenylpropionic acid (pHPPA), chavicol and Amplex red using Brucella melitensis antibody (BrAb) as a model analyte. The immunosensing body based on HRP-BrAb was constructed by dispersing graphite, BrAg and paraffin wax at room temperature. In a competitive immunoassay procedure, the BrAb competed with HRP-BrAb to react with the immobilized BrAg. In the enzymatic reaction, the binding HRP-BrAb on the sensing body surface can catalyze the polymerization reaction of TBG by H 2O2 forming fluorescent dimers and causing an increase in fluorescence intensity. TBG showed comparable ability for HRP detection and its enzyme-linked immunosensing reaction system, in a linear detection ranging of 3.5 ×10 -8 ~7.6 ×10 -6 g/L and with a detection limit of 1.7 ×10 -9 g/L. The immobilized biocomposite surface could be regenerated with excellent reproducibility (RSD=3.8%) by simply polishing with an alumina paper. The proposed immunosensing system has been used to determine the BrAb in rabbit serum samples with satisfactory results.

A capacitive chemical sensor for fenvalerate is reported. By using ac impedance measurements the sensor has been based on the decrease in capacitance caused by the analyte used as the template in the formulation of an electropolymerized molecularly imprinted polymer as receptor layer. Improvement of the insulating properties of the sensor was investigated in detail. The capacitive sensor was prepared by a deposition of a self-assembled monolayer of 2-mercaptobenzimidazole (2-MBI) before electropolymerization of 2-MBI and subsequent treatment with n-dodecanethiol to eliminate pinholes and defects in the polymerized 2-MBI film. From the calibration curve concentrations of fenvalerate up to 9 microg mL(-1) could be detected with a linear determination range up to 5 microg mL(-1) and a detection limit of 0.36 microg mL(-1). No significant interference was observed from common pyrethroid insecticides.

An amperometric metronidazole (MTZ) sensor using a glycosylated metalloporphyrin as a recognition element, which was incorporated in a carbon paste electrode, is reported. For the preparation of a MTZ-sensitive active material, 5, 10, 15, 20-tetrakis [2-(2, 3, 4, 6-tetraacetyl-β-D-glucopyranosyl)-1-O-phenyl]porphyrin (T(oglu) PPH2) and its Mn(III) complex MnT(o-glu)PPCl were synthesized from the reaction of pyrrole with ortho-acetylglycosylated benzaldehyde by Lindsay’s method. The MnT(oglu) PPCl-modified electrode showed excellent selectivity toward MTZ with respect to a number of interferents and exhibited stable response. The calibration graph obtained with the proposed sensor was linear over the range of 2.9×10-3-5.8×10-8 M/L, with a detection limit of 5.8×10-8 M/L for MTZ. Cyclic voltammetric measurements indicated that MnT(oglu) PPCl included in graphite-epoxy resin matrices could efficiently mediate electron transfer from the base electrode to MTZ causing a decrease of reduction potential for MTZ detection. The sensor could be regenerated by simply polishing with an alumina paper, with an excellent reproducibility (RSD=1.6%). The experimental conditions such as pH and applied working potential were optimized. The prepared sensor is applied for the determination of MTZ in pharmaceutical preparations and the results agreed with the values obtained by the pharmacopoeia method.

A novel amperometric immunosensor setup is described which uses horseradish peroxidase (HRP) as a label in conjunction with a current-based Brucella sensor. The Bacteria modified immunosensor was constructed by using a bio-composite formed by dispersing graphite powder into a mixture of Brucella melitensis and silicate polymer gel. The enzyme-labeled antibody can readily diffuse toward the encapsulated antigen (Brucella melitensis), which retains its binding properties, and the association reaction is easily detected at the surface exposed to the solution. The use of an o-aminophenol (o-AP) substrate and amperometric detection at -150 mV (vs. SCE) results in a relatively low detection limit of 3.5 ng/ml and a linear detection range of 3.5 ng/ml to 200 ng/ml. Based on an optimized parameter, the prepared sensor was used to detect the Brucella melitensis antibody in serum samples by using a competitive binding assay. The results demonstrate the feasibility of employing the proposed immunosensor for the detection for Brucella melitensis antibody in a clinical analysis.

A fluorescence sensor based on the supermolecular recognition by glycosylated metalloporphyrin for levamisole (LEV) assay is reported. For the preparation of a LEV-sensitive active material, 5, 10, 15, 20-tetrakis[2-(2, 3, 4, 6-tetraacetyl-β-D-glucopyranosyl)-1-O-phenyl] porphyrin and its metal complexes were synthesized and used in an optode membrane prepared by including glycosylated metalloporphyrin in chitosan matrice. The immobilized glycosylated metalloporphyrin is shown to be weakly fluorescent as a result of the inhibiting of the electron tansfer by central metal. The fluorescence enhancement of the metalloporphyrin modified optode membrane by LEV is based on the complexation with the central metal moiety of metalloporphyrin and weakening the inhibiting of the electron tansfer for metalloporphyrin. The glycosylated metalloporphyrin/chitosan optode membrane showed excellent selectivity toward LEV with respect to a number of interferents and exhibited stable response. The calibration graph obtained with the proposed sensor was linear over the range of 1.3 x 10 -5 -3.5 x 10 -7 M L -1 , with a detection limit of 3.5 x 10 -7 M L -1 for LEV. The prepared sensor is applied for the determination of LEV in pharmaceutical preparations and the results agreed with the values obtained by the pharmacopoeia method.