Highly sensitive, selective, and rapid fluorescence Hg2+ sensor based on DNA duplexes of poly(dT) and graphene oxide

Coupling T base with Hg(2+) to form stable T-Hg(2+)-T complexes represents a new direction in detection of Hg(2+). Here a graphene oxide (GO)-based fluorescence Hg(2+) analysis using DNA duplexes of poly(dT) that allows rapid, sensitive, and selective detection is first reported. The Hg(2+)-induced T(15)-(Hg(2+))(n)-T(15) duplexes make T(15) unable to hybridize with its complementary A(15) labelled with 6'-carboxyfluorescein (FAM-A(15)), which has low fluorescence in the presence of GO. On the contrary, when T(15) hybridizes with FAM-A(15) to form double-stranded DNA because of the absence of Hg(2+), the fluorescence largely remains in the presence of GO. A linear range from 10 nM to 2.0 μM (R(2) = 0.9963) and a detection limit of 0.5 nM for Hg(2+) were obtained under optimal experimental conditions. Other metal ions, such as Al(3+), Ag(+), Ca(2+), Ba(2+), Mg(2+), Zn(2+), Mn(2+), Co(2+), Pb(2+), Ni(2+), Cu(2+), Cd(2+), Cr(3+), Fe(2+), and Fe(3+), had no significant effect on Hg(2+) detection. Moreover, the sensing system was used for the determination of Hg(2+) in river water samples with satisfactory results.     

Colorimetric assay for mercury (II) based on mercury-specific deoxyribonucleic acid-functionalized gold nanoparticles

A colorimetric nanoprobe-mercury-specific DNA-functionalized gold nanoparticles (Au-MSD) was developed for sensing Hg(2+). The new mercury-sensing concept relies on measuring changes in the inhibition of "non-crosslinking" aggregation of Au-MSD-induced by the folding of mercury-specific DNA strand through the thymine-Hg(2+)-thymine (T-Hg(2+)-T) coordination. In the absence of Hg(2+), a high concentration of MgCl(2) (50 mM) results in a rapid aggregation of Au-MSD because of the removal of charge repulsion. When Hg(2+) is present, the particles remain stable due to the folding of MSD functionalized on the particle surface. The assay enables the colorimetric detection of Hg(2+) in the concentration range of 0.1-10 μM Hg(2+) ions with a detection limit of 60 nM, and allows for the selective discrimination of Hg(2+) ions from the other competitive metal ions. Toward the goal for practical applications, the sensor was further evaluated by monitoring Hg(2+) in fish tissue samples.     

Oligonucleotide-functionalized gold nanoparticles-enhanced QCM-D sensor for mercury(II) ions with high sensitivity and tunable dynamic range

A quartz crystal microbalance with dissipation monitoring (QCM-D) sensor was developed for highly sensitive and specific detection of mercury(II) ions (Hg(2+)) with a tunable dynamic range, using oligonucleotide-functionalized gold nanoparticles (GNPs) for both frequency and dissipation amplification. The fabrication of the sensor employed a 'sandwich-type' strategy, and formation of T-Hg(2+)-T structures in linker DNA reduced the hybridization of the GNPs-tagged DNA on the gold electrode, which could be used as the molecular switch for Hg(2+) sensing. This QCM-D mercury sensor showed a linear response of 10-200 nM, with detection limits of 4 nM and 7 nM for frequency and dissipation measurements, respectively. Moreover, the dynamic range of the sensor could be tuned by simply altering the concentration of linker DNA without designing new sensors in the cases where detection of Hg(2+) at different levels is required. This sensor afforded excellent selectivity toward Hg(2+) compared with other potential coexisting metal ions. The feasibility of the sensor was demonstrated by analyzing Hg(2+)-spiked tap- and lake-water samples with satisfactory recoveries. The proposed approach extended the application of the QCM-D system in metal ions sensing, and could be adopted for the detection of other analytes when complemented with the use of functional DNA structures.     

A simple and sensitive fluorescent sensing platform for Hg²+ ions assay based on G-quenching

In this work, a novel fluorescence biosensor was demonstrated for detection of Hg(2+) ions with relatively high selectivity and sensitivity. The sensing scheme was based on G-quenching induced by Hg(2+) ions. In the presence of Hg(2+) ions, the single-stranded signal probe which has carboxylfluorescein (FAM) and guanine segment at its 5' and 3' ends, respectively, folded into duplex-like structure via the Hg(2+)-mediated coordination of T-Hg(2+)-T base pairs. It brought guannine segment close to the dye and caused a remarkable decrease of fluorescence signal. The sensor showed a sensitive response to Hg(2+) ions in a concentration range from 0.5 to 10 μM, and a detection limit of 0.5 nM was given. This homogeneous system required only a single-labeled oligonucleotide, operated by concise procedures, and possessed comparable sensitivity as previous approaches. Furthermore, the sensor exhibits a great perspective for future practical applications.     

Control over surface DNA density on gold nanoparticles allows selective and sensitive detection of mercury(II)

We have developed a new highly selective and sensitive technique for the detection of Hg(2+) using DNA-functionalized gold nanoparticles (Au NPs) and OliGreen. This system is the first that allows the detection of Hg(2+) based on the release of DNA molecules, induced by conformational changes on Au NP surfaces, and its sensitivity is highly dependent upon surface DNA density. When Hg(2+) ions interact with the thymidine units of the DNA molecules bound to the Au NPs through Au-S bonds, the conformations of these DNA derivatives change from linear to hairpin structures, causing the release of some of the DNA molecules from the surface of the Au NPs into the bulk solution to react with OliGreen. The fluorescence of OliGreen-DNA complexes increased with increasing concentration of Hg(2+), and Hg(2+) could be detected at concentrations as low as 25 nM. A linear correlation existed between the fluorescence intensity and the concentration of Hg(2+) over the range 0.05-2.5 microM (R(2) = 0.98). This simple and cost-effective probe was applied to determine the spiked Hg(2+) in the pond samples; the recoveries (96-102%) suggested low matrix interference and high sensitivity.     

A simple and label-free sensor for mercury(II) detection in aqueous solution by malachite green based on a resonance scattering spectral assay

Mercury ions (Hg(2+)) can specifically interact with the thymine-rich Hg(2+) aptamer and malachite green (MG) to form the Hg(2+) aptamer-MG-Hg(2+) complex, inducing the increase of resonance scattering (RS) intensity at 611 nm, which enables the label-free detection of Hg(2+) in aqueous solution with high selectivity and a detection limit of 1.7 nM.     

Oligonucleotide-based label-free Hg2+ assay with a monomer-excimer fluorescence switch

A novel fluorescent Hg(2+) sensor was developed based on the T-Hg(2+)-T structure and a thioflavine T monomer-excimer fluorescent switch. Under optimum conditions, the selectivity is remarkably high, and Hg(2+) can be quantified over the dynamic range of 0.1 to 1.2 μM, with a limit of detection (LOD) of ~20 nM and a linear correlation coefficient of 0.995.     

Highly sensitive multiresponsive chemosensor for selective detection of Hg2+ in natural water and different monitoring environments

A new chemosensor RF1 that combines a ferrocene unit and a rhodamine block via the linkage of a carbohydrazone binding unit was designed and prepared for the highly selective detection of Hg (2+) in natural water. This chemosensor displays great brightness and fluorescence enhancement following Hg (2+) coordination within the limit of detection for Hg (2+) at 1 parts per billion (ppb). The fluorescence intensities are nearly proportional to the amount of Hg (2+) at the ppb level. It is capable of distinguishing between the safe and the toxic levels of inorganic mercury in drinking water. Hg (2+)-binding also arouses the absorption of the rhodamine moiety in RF1 significantly with the chromogenic detection limit for Hg (2+) at 50 ppb. The conventional UV-vis spectroscopic method thus has the potential to provide the critical information about the mercury hazard assessment for industrial wastewater discharging. The obvious and characteristic color change of the titration solution from colorless to pink upon the addition of Hg (2+) demonstrates that RF1 can be used for "naked-eye" detection of Hg (2+) in water. The Hg (2+) complexation also causes a significant shift of the redox potential about the ferrocene/ferrocenium couple. The electrochemical responses provide the possibility to quantitative analysis of Hg (2+) at the parts per million (ppm) level. Preliminary investigations in natural water samples including seawater and freshwater indicate that RF1 offers a direct and immediate Hg (2+) detection in complex media, pointing out its potential utility in environment monitoring and assessment. The responses of RF1 are Hg (2+) specific, and the chemosensor exhibits high selectivity toward Hg (2+) over other Group 12 metals, alkali, alkaline earth metals, and most of the divalent first-row transition metals. The RF1-Hg (2+) complex is successfully isolated and the Hg (2+)-binding is reversible. The crystal structure and spectral properties of its congener RF2 that contains one ferrocene group and two rhodamine 6G moieties were also investigated for a comparison.     

Rhodamine hydrazone derivatives as Hg2+ selective fluorescent and colorimetric chemosensors and their applications to bioimaging and microfluidic system

In this paper, we report new rhodamine hydrazone derivatives bearing thiol and carboxylic acid groups as selective fluorescent and colorimetric chemosensors for Hg(2+). The ring-opening process of spirolactam enables the large fluorescent enhancement and colorimetric change upon the addition of Hg(2+). The sample containing Hg(2+) was mixed with one of the chemosensors in a microchannel where the sensor was examined using confocal laser scanning microscopy. A plot of the fluorescent intensities of both chemosensors versus the log concentration of Hg(2+) exhibited a linear response (r(2)=0.95) in the range of 1 nM-1 μM, and the detection limits were 1 nM and 4.2 nM, respectively. Both chemosensors also enable the visualization of Hg(2+) accumulated in the nematode Caenorhabditis elegans previously exposed to nanomolar concentrations of Hg(2+).     

Amplified fluorescence detection of mercury(II) ions (Hg2+) using target-induced DNAzyme cascade with catalytic and molecular beacons

In this work, a fluorescent sensing strategy was developed for the detection of mercury(II) ions (Hg(2+)) in aqueous solution with excellent sensitivity and selectivity using a target-induced DNAzyme cascade with catalytic and molecular beacons (CAMB). In order to construct the biosensor, a Mg(2+)-dependent DNAzyme was elaborately designed and artificially split into two separate oligonucleotide fragments. In the presence of Hg(2+), the specific thymine-Hg(2+)-thymine (T-Hg(2+)-T) interaction induced the two fragments to produce the activated Mg(2+)-dependent DNAzyme, which would hybridize with a hairpin-structured MB substrate to form the CAMB system. Eventually, each target-induced activated DNAzyme could catalyze the cleavage of many MB substrates through true enzymatic multiple turnovers. This would significantly enhance the sensitivity of the Hg(2+) sensing system and push the detection limit down to 0.2 nM within a 20 min assay time, much lower than those of most previously reported fluorescence assays. Owning to the strong coordination of Hg(2+) to the T-T mismatched pairs, this proposed sensing system exhibited excellent selectivity for Hg(2+) detection, even in the presence of 100 times of other interferential metal ions. Furthermore, the applicability of the biosensor for Hg(2+) detection in river water samples was demonstrated with satisfactory results. These advantages endow the sensing strategy with a great potential for the simple, rapid, sensitive, and specific detection of Hg(2+) from a wide range of real samples.     

Visual and on-site detection of mercury(II) ions on lateral flow strips using DNA-functionalized gold nanoparticles

A test strip for detection of Hg(2+) in aqueous solution based on the DNA-functionalized gold nanoparticles (DNA-AuNPs) was developed and evaluated. When Hg(2+) ions were introduced, the biotinylated DNA(2) hybridized with thiolated DNA(1) functionalized on the AuNPs (DNA(1)-AuNPs) to form mismatch complexes through thymine-Hg(2+)-thymine (T-Hg(2+)-T) coordination. The formed mismatch complexes and excess DNA(1)-AuNPs could be captured on the test line formed by streptavidin and the control line formed by DNA(3)-BSA, respectively. Two red lines appeared due to the accumulation of AuNPs, enabling visual detection of Hg(2+) with a detection limit of about 6 nM. The assay results can be obtained within 5 min. The results show that the test strip has excellent sensitivity and selectivity for detection of Hg(2+); thus it holds a great potential for rapid, on-site and real time detection of Hg(2+).     

A one-step colorimetric method of analysis detection of Hg2+ based on an in situ formation of Au@HgS core-shell structures

A new approach for the detection of Hg(2+) is reported based on color changes from which gold nanoparticles (Au NPs) are surrounded by a layer of HgS quantum dots to form in situ Au@HgS core-shell nanostructures. The surface plasmon resonance (SPR) absorption of the gold core was changed due to a shell layer of HgS formed on the surface of the Au NPs, which brings the colour change of the aqueous solution. Therefore, Hg(2+) can be recognized by visualizing the colour change of the Au@HgS core-shell nanostructures, and can be detected quantitatively by measurement of the UV-vis spectra. Some effects on the detection of Hg(2+) were investigated in detail. This method was used to detect Hg(2+) with excellent selectivity and high sensitivity. In our method, the lowest detected concentrations for mercury ions were 5.0 × 10(-6) M observed by the naked eye and 0.486 nM as measured by UV-vis spectra. At the range from 8.0 × 10(-5) to 1.0 × 10(-8) M of Hg(2+), this method was shown to have a good linear relationship.     

Label-free DNA hybridization detection and single base-mismatch discrimination using CE-ICP-MS assay

Detecting a specific DNA sequence and discriminating single base-mismatch is critical to clinical diagnosis, paternity testing, forensic sciences, food and drug industry, pathology, genetics, environmental monitoring, and anti-bioterrorism. To this end, capillary electrophoresis (CE) coupled with the inductively coupled plasma mass spectrometry (ICP-MS) method is developed using the displacing interaction between the target ssDNA and the competitor Hg(2+) for the first time. The thymine-rich capture ssDNA 1 is interacted with the competitor Hg(2+), forming an assembled complex in a hairpin-structure between the thymine bases arrangement at both sides of the capture ssDNA 1. In the presence of a target ssDNA with stronger affinity than that of the competitor Hg(2+), the energetically favorable hybridization between capture ssDNA 1 and the target ssDNA destroys the hairpin-structure and releases the competitor as free Hg(2+), which was then read out and accurately quantified by CE-ICP-MS assay. Under the optimal CE separation conditions, free Hg(2+) ions and its capture ssDNA 1 adduct were baseline separated and detected on-line by ICP-MS; the increased peak intensity of free Hg(2+) against the concentration of perfectly complementary target ssDNA was linear over the concentration range of 30-600 nmol L(-1) with a limit of detection of 8 nmol L(-1) (3s, n = 11) in the pre-incubated mixture containing 1 μmol L(-1) Hg(2+) and 0.2 μmol L(-1) capture ssDNA 1. This new assay method is simple in design since any target ssDNA binding can in principle result in free Hg(2+) release by 6-fold Hg(2+) signal amplification, avoiding oligonucleotide labeling or assistance by excess signal transducer and signal reporter to read out the target. Due to element-specific detection of ICP-MS in our assay procedure, the interference from the autofluorescence of substrata was eliminated.     

An efficient rhodamine thiospirolactam-based fluorescent probe for detection of Hg2+ in aqueous samples

This paper described the optimized design, synthesis and application of a novel rhodamine thiospirolactam derivative as an 'off-on' fluorescent probe for the detection of Hg(2+) in aqueous samples. The 'off-on' fluorescence and color signal change of the probe is based on an Hg(2+)-triggered domino reaction which brings on the opened-ring form of the rhodamine spirolactam to regain the conjugated system of the rhodamine skeleton. In the well designed probe, the thiospirolactam serves as both Hg(2+) binding unit and electron-defect carbon centre, a phenolic hydroxyl with very strong nucleophilicity after deprotonation is chosen as the attacking unit, and a benzene ring is introduced on the linker to afford steric effects, which benefits an efficient nucleophilic reaction, with a high sensitivity towards Hg(2+). It exhibits a stable response for Hg(2+) from 1.0 × 10(-8) to 1.0 × 10(-6) M, with a detection limit of 3.0 × 10(-9) M. The response of the probe to Hg(2+) is highly selective and pH-insensitive, with a fast response time. All these unique features make it particularly favorable for cellular Hg(2+) imaging applications. It has been preliminarily used for highly sensitive monitoring of Hg(2+) levels in living cells with satisfying resolution.     

Fluorescent gold clusters as nanosensors for copper ions in live cells

This paper reports the use of fluorescent gold nanoclusters synthesized using bovine serum albumin (Au-BSA) for the sensing of copper ions in live cells. The fluorescence of the clusters was found to be quenched by Cu(2+) enabling its detection in cells. The selectivity of the nanosensor was demonstrated in the presence of several cations excluding Hg(2+). We did not study the effect of Hg(2+) since it was reported earlier. The present study suggests that Cu(2+) induced fluorescence quenching is due to its binding to BSA rather than the fluorescence quenching by metal-metal interaction as in the case of Hg(2+). The Au-BSA showed excellent selectivity to Cu(2+) at various pH conditions. The 'turn off' of fluorescence can be retrieved by a Cu(2+) chelator glycine. Our results showed that gold clusters can be used as a 'turn off' sensor for copper and a 'turn on' sensor for glycine. Under the experimental conditions, the probe showed a response for Cu(2+) over a range of 100 μM to 5 mM with a detection limit of 50 μM. The role of Cu(2+) in the misfolding and disassembly of Prion Protein (PrP) leading to various maladies is well ascertained. The methodology we reported here seems to be useful in supplementing other techniques in predicting disease conditions involving Cu(2+).     

Novel hemicyanine dye as colorimetric and fluorometric dual-modal chemosensor for mercury in water

A novel water soluble Hg(2+)-selective chemosensor 1 with hemicyanine as fluorescent reporting group and NO(2)Se(2) chelating unit as ion binding site was reported. Chemosensor 1 shows a specific Hg(2+) selectivity and discrimination between Hg(2+) and chemically similar ions in conjunction with a visible colorimetric change from red to colorless, potentially leading to both "naked-eye" and fluorometric detection of Hg(2+) cations.     

A blue fluorescent antibody-cofactor sensor for mercury

[reaction: see text] A chemically programmed antibody sensor, consisting of a stilbenyl boronic acid cofactor and monoclonal antibody EP2-19G2, provides a new method of mercury detection. The fluorescent antibody sensor generates an intense powder blue fluorescence when bound to the stilbenyl boronic acid cofactor; however, it is quenched in the presence of Hg(2+) ions. The EP2-19G2-cofactor biosensor provides micromolar sensitivity and selectivity toward Hg(2+) ions over a wide range of metal ions in aqueous solution.     

Hg2+-mediated aggregation of gold nanoparticles for colorimetric screening of biothiols

In this work, we report a colorimetric assay for the screening of biothiols including glutathione (GSH), cysteine (Cys), and homocysteine (Hcys) based on Hg(2+)-mediated aggregation of gold nanoparticles (AuNPs). Hg(2+) can induce aggregation of thiol-containing naphthalimide (1) capped AuNPs due to the cross-linking interactions from the resulting "thymine-Hg(2+)-thymine" (T-Hg(2+)-T) analogous structure. When Hg(2+) is firstly treated with biothiols, followed by mixing with 1-capped AuNPs suspension, AuNPs undergo a transformation from an aggregation to a dispersion state depending on the concentration of biothiols. This anti-aggregation or re-dispersion of AuNPs is due to the higher affinity of Hg(2+) for biothiols relative to compound 1. The corresponding color variation in the process of anti-aggregation of AuNPs can be used for the quantitative screening of biothiols through UV-vis spectroscopy or by the naked eye. Under optimized conditions, a good linear relationship in the range of 0.025-2.28 μM is obtained for GSH, 0.035-1.53 μM for Cys, and 0.040-2.20 μM for Hcys. The detection limits of this assay for GSH, Cys, and Hcys are 17, 9, and 18 nM, respectively. This colorimetric assay exhibits a high selectivity and sensitivity with tunable dynamic range. The proposed method has been successfully used in the determination of total biothiol content in human urine samples.     

Ratiometric detection of Cr3+ and Hg2+ by a naphthalimide-rhodamine based fluorescent probe

Newly synthesized rhodamine derivatives, L(1) and L(2), are found to bind specifically to Hg(2+) or Cr(3+) in presence of large excess of other competing ions with associated changes in their optical and fluorescence spectral behavior. These spectral changes are significant enough in the visible region of the spectrum and thus, allow the visual detection. For L(1), the detection limit is even lower than the permissible [Cr(3+)] or [Hg(2+)] in drinking water as per standard U.S. EPA norms; while the receptor, L2 could be used as a ratiometric sensor for detection of Cr(3+) and Hg(2+) based on the resonance energy transfer (RET) process involving the donor naphthalimide and the acceptor Cr(3+)/Hg(2+)-bound xanthene fragment. Studies reveal that these two reagents could be used for recognition and sensing of Hg(2+)/Cr(3+). Further, confocal laser microscopic studies confirmed that the reagent L(2) could also be used as an imaging probe for detection of uptake of these ions in A431 cells.     

A versatile water soluble fluorescent probe for ratiometric sensing of Hg2+ and bovine serum albumin

A novel tetraazamacrocycle fluorescent sensor (6-(1-(dimethylamino)-5-naphthalene sulfonyl)-3,6,9,15-tetraazabicyclo[9.3.1] pentadeca-1(15),11,13-triene, 1) has been designed and prepared, which can be utilized for selective and ratiometric sensing of Hg(2+) and bovine serum albumin (BSA) with two different responsive modes in aqueous solution at physiological pH (50 mM Tris-HCl, pH 7.6). Above 0.5 ppb Hg(2+) can be discerned by coordination with 1 and the emission color changes enable 1 to be applied to a fast Hg(2+) test paper assay. Sensor 1 has also been demonstrated to be easily cell-penetrable and applicable for Hg(2+) imaging in living cells. Imaging of BSA in the gel using SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) stained in the medium containing 1 verified that the binding of 1 and BSA was successful in the presence of nonprotein substances. The linear range of 1 towards BSA utilizing ratiometric fluorescent calibration via noncovalent interaction in solution is 0-100 μg mL(-1) with a detection limit of 1 μg mL(-1), and has been successfully employed to determine the albumin concentration in blood serum by means of ratiometric fluorescent measurements for the first time. Finally, sensor 1 behaves as a fluorescent molecular switch composed of triple logic gates upon chemical inputs of Hg(2+) and BSA, which potentially provides intelligent diagnostics for Hg(2+) contaminated serum on the nanoscale.