Ratiometric fluorescence chemosensors for copper(II) and mercury(II) based on FRET systems

A system based on FRET mechanism, comprising a coumarin donor and a rhodamine acceptor, was developed for the selective and quantitative detection of metal ions. Fluorescent chemosensors RCs, linked by 1,2-diethylamine, exhibit significant fluorescence enhancement and excellent selectivity toward Cu²⁺. Fluorescent probes CRB and CR6G, linked by hydrazide, function as ratiometric receptors for Cu²⁺ chromogentically and fluorogentically in organic-aqueous media. Furthermore, the characteristic rhodamine-based fluorescence response of CRB (excitation at 550 nm) exhibits high selectivity for Hg(II). The construction of this kind of universal FRET system opens a broader prospect for future design of ratiometric fluorescent probes.     

基于聚合物核壳纳米粒子的Hg~(2+)比率型荧光检测传感器

采用一步聚合的方法,制备了以疏水的聚甲基丙烯酸甲酯(PMMA)为核、亲水性的聚电解质支化聚乙烯亚胺(PEI)为壳的纳米粒子分散液.将供体荧光团4-胺基-7-硝基-N-辛基苯并[1,2,5]噁二唑(NBD)以包埋的方式在聚合过程中直接引入PMMA核内部,而受体荧光团罗丹明衍生物SRHB通过吸附作用进入PEI-PMMA核壳界面,构成了含有两种不同荧光分子且可对Hg2+进行荧光比率检测的传感器.考察了含荧光分子的聚合物粒子光谱学性质,证明两种荧光分子均被引入了聚合物粒子体系.在汞离子的荧光检测试验中,加入Hg2+后,体系中的NBD荧光强度下降,而罗丹明的特征发射峰在579 nm处出现,并随着Hg2+浓度的增加,受体/供体的荧光强度比值呈现增长趋势.研究还发现,聚合物粒子基荧光探针对于Hg2+具有较好的选择性,且最佳使用范围是体系pH值在5～8之间,其检测Hg2+的最低浓度可达到1μmol/L.     

Ratiometric fluorescence probe for two-photon bioimaging of Cr in living cells

We presented a ratiometric fluorescent probe dansylamide–rhodamine dyad (DANS–RB) for selectively detecting Cr³⁺ in semi-aqueous solution. The detection mechanism relies on the fluorescent resonance energy transfer (FRET) process from the dansylamide (energy donor) to the rhodamine (energy acceptor) after the addition of Cr³⁺. The cell-permeability of DANS–RB was confirmed by the two-photon fluorescence microscopy experiments, which demonstrated DANS–RB was a good candidate for monitoring the intracellular Cr³⁺ level with the ratiometric fluorescent method. Combining the excellent selectivity, the ratiometric quantitative detection, and the cell-permeability, DANS–RB may find a broad application in the investigation on biologically relevant species in living cells.     

Fluorescence energy transfer-based multiplexed hybridization assay using gold nanoparticles and quantum dot conjugates on photonic crystal beads

A multiplexed assay strategy was developed for the detection of nucleic acid hybridization. It is based on fluorescence resonance energy transfer (FRET) between gold nanoparticles (AuNPs) and multi-sized quantum dots (QDs) deposited on the surface of silica photonic crystal beads (SPCBs). The SPCBs were first coated with a three-layer primer film formed by the alternating adsorption of poly(allylamine hydrochloride) and poly(sodium 4-styrensulfonate). Probe DNA sequences were then covalently attached to the carboxy groups at the surface of the QD-coated SPCBs. On addition of DNA-AuNPs and hybridization, the fluorescence of the donor QDs is quenched because of the close proximity of the AuNPs. However, the addition of target DNA causes a recovery of the fluorescence of the QD-coated SPCBs, thus enabling the quantitative assay of hybridized DNA. Compared to fluorescent dyes acting as acceptors, the use of AuNPs results in much higher quenching efficiency. The multiplexed assay displays a wide linear range, high sensitivity, and very little cross-reactivity. This work, where such SPCBs are used for the first time in a FRET assay, is deemed to present a new and viable approach towards high-throughput multiplexed gene assays.

Selective solid-phase extraction of trace mercury(II) using a silica gel modified with diethylenetriamine and thiourea

We describe a solid phase extractor for selective separation and preconcentration of Hg(II) ion. It was prepared by immobilizing the adduct of diethylenetriamine and thiourea on silica gel. The effects of solution acidity, preconcentration time, sample flow rate and volume were optimized. The results show that Hg(II) can be selectively extracted from acidic solutions and in presence of common other metal ions. The adsorbent is stable, can be reused more than 10 times, and the maximum adsorption capacity is 23 mg g^?1. Hg(II) was quantified by inductively coupled plasma optical emission spectrometry. The method has a detection limit of 23 ng L^?1, and the relative standard deviation is <2 %. The procedure was validated by analyzing two standard materials (river sediment and hair powder), and was successfully applied to the preconcentration of Hg(II) in real samples.

A FRET system built on quartz plate as a ratiometric fluorescence sensor for mercury ions in water

Due to the hazardous nature of mercury ions, the development of a cost effective, sensitive and field-portable sensor is of high significance for both industry and civilian use. In this work, a FRET-based ratiometric sensor for detecting mercury ions in water was fabricated by depositing a multilayered silica structure on a quartz plate. For the preparation of the film-based sensor, a silica support layer was first deposited on the quartz plate by using the sol-gel spin-coating procedure, and three ultrathin functional layers (donor, spacer and receptor) were then deposited on the support layer by dip-coating in a stepwise manner in toluene solution. As the film-based sensor was placed into an aqueous solution of Hg(2+), the non-fluorescent receptor (a spirolactam rhodamine derivative) on the film surface could form a complex with the mercury ion and act as the acceptor of the energy transfer. Upon excitation, the donor (a nitrobenzoxadiazolyl derivative, NBD) could transfer its excited energy from the donor layer to the acceptor on the film surface via the 'through space' energy transfer process, thus realizing the FRET-based ratiometric sensing for mercury ions. The sensor can selectively detect Hg(2+) in water with the detection limit of 1 μM. This solid film sensor is capable of being easily-portable and visualized detection. This strategy may offer new approaches for constructing other FRET-based solid-state devices.     

Detection of Helicobacter pylori with a nanobiosensor based on fluorescence resonance energy transfer using CdTe quantum dots

We report on a method for the sensitive determination of Helicobacter that is based on fluorescence resonance energy transfer using two oligonucleotide probes labeled with CdTe quantum dots (QDs) and 5-carboxytetramethylrhodamine (Tamra) respectively. QDs labeled with an amino-modified first oligonucleotide, and a Tamra-labeled second oligonucleotide were added to the DNA targets upon which hybridization occurred. The resulting assembly brings the Tamra fluorophore (the acceptor) and the QDs (the donor) into close proximity and causes fluorescence resonance energy transfer (FRET) to occur upon photoexcitation of the donor. In the absence of target DNA, on the other hand, the probes are not ligated, and no emission by the Tamra fluorophore is produced due to the lack of FRET. The feasibility of the method was demonstrated by the detection of a synthetic 210-mer nucleotide derived from Helicobacter on a nanomolar level. This homogeneous DNA detection scheme is simple, rapid and efficient, does not require excessive washing and separation steps, and is likely to be useful for the construction of a nanobiosensor for Helicobacter species.

A FRET-based ratiometric sensor for mercury ions in water with multi-layered silica nanoparticles as the scaffold

A FRET system was built for ratiometric sensing of Hg(2+) in water with multilayered silica nanoparticles as the scaffold. This architecture ensures the control over the location of both donor and acceptor and their separation distance within nanoparticles, affording higher energy transfer efficiency and higher signal-to-background ratio for particle-based sensing.     

Colloidal silica beads modified with quantum dots and zinc (II) tetraphenylporphyrin for colorimetric sensing of ammonia

Colloidal crystal beads (CCBs) were fabricated by assembling monodisperse silica nanoparticles via a microfluidic device. The pore size of the CCBs was tuned by using different nanoparticles. The CCBs were then coated with cadmium telluride quantum dots and zinc(II) meso-tetraphenylporphyrin for the purpose of optical sensing. Ammonia causes the color of the sensor to change from green to red. The method has a dynamic range of 0–2500 ppm, good reversibility, and is not sensitive to humidity. The limit of detection is 7 ppm. The sensor has the advantage of a porous microcarrier structure and that pore sizes can be well controlled and thus can fulfill various demands in gas detection.

A FRET fluorescent chemosensor SPAQ for Zn based on a dyad bearing spiropyran and 8-aminoquinoline unit

A novel FRET fluorescent sensor SPAQ containing 8-aminoquinoline (donor) and spiropyran derivative (acceptor) was designed and synthesized for detecting Zn²⁺. The probe successfully exhibited a fluorescence turn on and ratiometric response for Zn²⁺ in ethanol solution with high selectivity. Upon excitation at 370 nm, the modulation of the emission intensity of SPAQ at 645 and 470 nm was achieved in the presence of Zn²⁺ by fluorescent resonance energy transfer (FRET) and chelation-enhanced fluorescence (CHEF) effects.     

Preparation of liposomes loaded with quantum dots, fluorescence resonance energy transfer studies, and near-infrared in-vivo imaging of mouse tissue

We report on a simple, fast and convenient method to engineer lipid vesicles loaded with quantum dots (QDs) by incorporating QDs into a vesicle-type of lipid bilayer using a phase transfer reagent. Hydrophilic CdTe QDs and near-infrared (NIR) QDs of type CdHgTe were incorporated into liposomes by transferring the QDs from an aqueous solution into chloroform by addition of a surfactant. The QD-loaded liposomes display bright fluorescence, and the incorporation of the QDs into the lipid bilayer leads to enhanced storage stability and reduced sensitivity to UV irradiation. The liposomes containing the QD were applied to label living cells and to image mouse tissue in-vivo using a confocal laser scanning microscope, while NIR images of mouse tissue were acquired with an NIR fluorescence imaging system. We also report on the fluorescence resonance energy transfer (FRET) that occurs between the CdTe QDs (the donor) and the CdHgTe QDs (the acceptor), both contained in liposomes. Based on these data, this NIR FRET system shows promise as a tool that may be used to study the release of drug-loaded liposomes and their in vivo distribution.

Aptamer based test stripe for ultrasensitive detection of mercury(II) using a phenylene-ethynylene reagent on nanoporous silver as a chemiluminescence reagent

We describe a paper-based chemiluminescence (CL) test for the determination of mercury(II) ion. A single-stranded DNA aptamer was first covalently immobilized via its amino groups to the hydroxy groups on the surface of cellulosic paper. The aptamer probes can capture Hg(II) ions due to their specific interaction with thymine. The CL reagent (a caboxylated phenylene-ethynylene referred to as P-acid) was immobilized on nanoporous silver (NPS@P-acid) and used a CL label on the aptamer. The stripe is then contacted with a sample containing Hg(II) ions and CL is induced by the addition of permanganate. CL intensity depends on the concentration of Hg(II) because Hg(II) increases the quantity of the P-acid-conjugated aptamer. The highly active surface of the NPS@P-acid composites results in an 8-fold higher CL intensity compared to the use of pure P-acid. This enables Hg(II) ion to be quantified in the 20 nM to 0.5 μM concentration range, with a limit of detection as low as 1 pM. This CL aptasensor is deemed to represent a promising tool for simple, rapid, and sensitive detection of Hg(II).

Fluorescent probe for copper(II) ion based on a rhodamine spirolactame derivative, and its application to fluorescent imaging in living cells

A fluorescent probe for Cu(II) ion is presented. It is based on the rhodamine fluorophore and exhibits high selectivity and sensitivity for Cu(II) ion in aqueous methanol (2:8, v/v) at pH 7.0. The response is based on a ring opening reaction and formation of a strongly fluorescent 1:1 complex. The response is reversible and linear in the range between 50?nM and 900?nM, with a detection limit of 7.0?nM. The probe was successfully applied to fluorescent imaging of Cu(II) ions in HeLa cells.

Solid-phase extraction of mercury(II) with magnetic core-shell nanoparticles, followed by its determination with a rhodamine-based fluorescent probe

Magnetic Fe₃O₄@SiO₂ core shell nanoparticles containing diphenylcarbazide in the shell were utilized for solid phase extraction of Hg(II) from aqueous solutions. The Hg(II) loaded nanoparticles were then separated by applying an external magnetic field. Adsorbed Hg(II) was desorbed and its concentration determined with a rhodamine-based fluorescent probe. The calibration graph for Hg(II) is linear in the 60 nM to 7.0 μM concentration range, and the detection limit is at 23 nM. The method was applied, with satisfying results, to the determination of Hg(II) in industrial waste water.

Highly sensitive FRET-based fluorescence immunoassay for aflatoxin B1 using cadmium telluride quantum dots

We report on a competitive immunoassay for the determination of aflatoxin B1 using fluorescence resonance energy transfer (FRET) from anti-aflatoxin B1 antibody (immobilized on the shell of CdTe quantum dots) to Rhodamine 123 (Rho 123-labeled aflatoxin B1 bound to albumin). The highly specific immunoreaction between the antibody against aflatoxin B1 on the QDs and the labeled-aflatoxin B1 brings the Rho 123 fluorophore (acting as the acceptor) and the QDs (acting as the donor) in close spatial proximity and causes FRET to occur upon photoexcitation of the QDs. In the absence of unlabeled aflatoxin B1, the antigen-antibody complex is stable, and strong emission resulting from the FRET from QDs to labeled aflatoxin B1 is observed. In the presence of aflatoxin B1, it will compete with the labeled aflatoxin B1-albumin complex for binding to the antibody-QDs conjugate so that FRET will be increasingly suppressed. The reduction in the fluorescence intensity of the acceptor correlates well with the concentration of aflatoxin B1. The feasibility of the method was established by the detection of aflatoxin B1 in spiked human serum. There is a linear relationship between the increased fluorescence intensity of Rho 123 with increasing concentration of aflatoxin B1 in spike human serum, over the range of 0.1–0.6 μmol·mL^?1. The limit of detection is 2?×?10^?11 M. This homogeneous competitive detection scheme is simple, rapid and efficient, and does not require excessive washing and separation steps.

Core-shell fluorescent silica nanoparticles for sensing near-neutral pH values

pH-responsive fluorescent core-shell silica nanoparticles (SiNPs) were prepared by encapsulating the pH-sensitive fluorophore 8-hydroxypyrene-1,3, 6-trisulfonate into their silica shell via a facile reverse microemulsion method. The resulting SiNPs were characterized by SEM, TEM, fluorescence lifetime spectroscopy, photobleaching experiments, and photoluminescence. The core-shell structure endows the SiNPs with reduced photobleaching, excellent photostability, minimized solvatachromic shift, and increased fluorescence efficiency compared to the free fluorophore in aqueous solution. The dynamic range for sensing pH ranges from 5.5 to 9.0. The nanosensors show excellent stability, are highly reproducible, and enable rapid detection of pH. The results obtained with the SiNPs are in good agreement with data obtained with a glass electrode.

Synthesis of ratiometric fluorescent nanoparticles for sensing oxygen

A facile reprecipitation-encapsulation method is used for the preparation of ratiometric fluorescent nanoparticles (NPs) for sensing intracellular oxygen. The surface of the NPs is modified in-situ with poly-L-lysine, which renders good biocompatibility and enables easy internalization into living cells. The sensor NPs contain a red fluorescent probe whose fluorescence is sensitive to oxygen with a quenching response of 77 % on going from nitrogen saturation to oxygen saturation, and a reference dye giving a green signal that acts as an oxygen-independent reference. The ratio of the two emissions serves as the analytical information and is sensitive to dissolved oxygen in the 0–43?ppm concentration range. When incorporated into cells, the ratio of the signals increases by 400?% on going from oxygen-saturated to oxygen-free environment.

A colorimetric probe for the rapid and selective determination of mercury(II) based on the disassembly of gold nanorods

We report on a novel mercury(II)-controlled approach for the disassembly of gold nanorods (AuNRs) that has led to a detection system for Hg(II). The modified AuNRs were fabricated by functionalizing AuNRs with L-cysteine via a thiol group chemisorption-type of interaction. L-cysteine induces the assembly of AuNRs through cooperative electrostatic interaction upon which the color of the solution of the AuNRs changes from blue-green to gray dark. The addition of Hg(II), in turn, causes the disassembly of the modified AuNRs and the color of the solution returns to blue-green. This effect enables the optical determination of Hg(II) in aqueous solution, with a linear response in the 0.5 to 250 μM Hg(II) concentration range, an excellent selectivity for Hg(II), and with recoveries ranging from 99 % to 106 % in spiked environmental water samples.

A nanoparticle-supported fluorescence resonance energy transfer system formed via layer-by-layer approach as a ratiometric sensor for mercury ions in water

This article describes the design and preparation of a novel fluorescence resonance energy transfer (FRET)-based ratiometric sensor with the polymer nanoparticle as scaffold for detecting Hg²⁺ in aqueous media. In this study, a fluorescent dye fluorescein isothiocyanate (FITC, served as the donor) and a spirolactam rhodamine derivative (SRHB, served as mercury ion probe) were covalently attached onto polyethylenimine (PEI) and polyacrylic acid (PAA) respectively; and a ratiometric sensing system was then formed through the deposition of the donor- and probe-containing polyelectrolytes onto the negatively charged polymer particles via the layer-by-layer approach. The ratiometric fluorescent signal change of the system is based on the intra-particle fluorescence resonance energy transfer (FRET) process modulated by mercury ions. Under optimized structural and experimental conditions, the particle-based detection system exhibits stable response for Hg²⁺ in aqueous media. More importantly, in this newly developed particle-based detection system formed by LBL approach, varied numbers of the PAA/PEI layers which served as the spacer could be placed between the donor-containing layer and the probe-containing layer, hence the donor–acceptor distance and energy transfer efficiency could be effectively tuned (from ca. 25% to 76%), this approach has well solved the problem for many particle-based FRET systems that the donor–acceptor distance cannot be precisely controlled. Also, it is found that the ratiometric sensor is applicable in a pH range of 4.6–7.3 in water with the detection limit of 200 nM. This approach may provide a new strategy for ratiometric detection of analytes in some environmental and biological applications.     

Preconcentration of mercury(II) using a thiol-functionalized metal-organic framework nanocomposite as a sorbent

A novel type of porous metal-organic framework (MOF) was obtained from thiol-modified silica nanoparticles and the copper(II) complex of trimesic acid. It is shown that this nanocomposite is well suitable for the preconcentration of Hg(II) ions. The nanocomposite was characterized by Fourier transfer infrared spectroscopy, X-ray powder diffraction, energy-dispersive X-ray diffraction and scanning electron microscopy. The effects of pH value, sorption time, elution time, the volume and concentration of eluent were investigated. Equilibrium isotherms were studied, and four models were applied to analyze the equilibrium adsorption data. The results revealed that the adsorption process obeyed the Langmuir model. The maximum monolayer capacity and the Langmuir constant are 210 mg g^?1 and 0.273 L mg^?1, respectively. The new MOF-based nanocomposite is shown to be an efficient and selective sorbent for Hg(II). Under the optimal conditions, the limit of detection is 20 pg mL^?1 of Hg(II), and the relative standard deviation is <7.2 % (for n?=?3). The sorbent was successfully applied to the rapid extraction of Hg(II) ions from fish, sediment, and water samples.