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.

Quantum dots on electrodes—new tools for bioelectroanalysis

The review covers recent developments in which quantum dots (QDs) are combined with electrodes for detection of analytes. Special focus will be on the generation of photocurrents and the possibility of spatially resolved, light-directed analysis. Different modes for combining biochemical reactions with QDs will be discussed. Other applications involve the use of QDs as labels in binding analysis. Different methods have been developed for read-out. In addition to photocurrent analysis, voltammetric detection of metals and electrochemiluminescence (ECL) can be used. In the latter, light is the sensor signal. ECL-based systems combine the advantage of very sensitive analytical detection with rather simple instrumentation.

In vitro and in vivo imaging with quantum dots

Quantum dots (QDs), also named semiconductor nanocrystals, have initiated a new realm of bioscience by combining nanomaterials with biology, which will profoundly influence future biological and biomedical research. In this review, we describe the extraordinary optical properties of QDs and developments in methods for their synthesis. We focus on fluorescent imaging with QDs both in vitro and in vivo, and the cytotoxicity of QDs and potential barriers to their use in practical biomedical applications. Finally, we provide insights into improvements aimed at decreasing the cytotoxicity of QDs and the future outlook of QD applications in biomedical fields.

Use of quantum dots in the development of assays for cancer biomarkers

Biomarker assays may be useful for screening and diagnosis of cancer if a set of molecular markers can be quantified and statistically differentiated between cancerous cells and healthy cells. Markers of disease are often present at very low concentrations, so methods capable of low detection limits are required. Quantum dots (QDs) are nanoparticles that are emerging as promising probes for ultrasensitive detection of cancer biomarkers. QDs attached to antibodies, aptamers, oligonucleotides, or peptides can be used to target cancer markers. Their fluorescent properties have enabled QDs to be used as labels for in-vitro assays to quantify biomarkers, and they have been investigated as in-vivo imaging agents. QDs can be used as donors in assays involving fluorescence resonance energy transfer (FRET), or as acceptors in bioluminescence resonance energy transfer (BRET). The nanoparticles are also capable of electrochemical detection and are potentially useful for “lab-on-a-chip” applications. Recent developments in silicon QDs, non-blinking QDs, and QDs with reduced-size and controlled-valence further make these QDs bioanalytically attractive because of their low toxicity, biocompatibility, high quantum yields, and diverse surface modification flexibility. The potential of multiplexed sensing using QDs with different wavelengths of emission is promising for simultaneous detection of multiple biomarkers of disease.

Method for determination of microcystin-leucine-arginine in water samples based on the quenching of the fluorescence of bioconjugates between CdSe/CdS quantum dots and microcystin-leucine-arginine antibody

We have developed a method for the determination of microcystin-leucine-arginine (MC-LR) in water samples that is based on the quenching of the fluorescence of bioconjugates between CdSe/CdS quantum dots (QDs) and the respective antibody after binding of MC-LR. The core-shell CdSe/CdS QDs were modified with 2-mercaptoacetic acid to improve water solubility while their high quantum yields were preserved. Monoclonal MC-LR antibody was then covalently bioconjugated to the QDs. It was found that the fluorescence intensity of the bioconjugates was quenched in the presence of MC-LR. A linear relationship exists between the extent of quenching and the concentration of MC-LR. Parameters affecting the quenching were investigated and optimized. The limit of detection is 6.9?×?10^?11 mol L^?1 (3σ). The method was successfully applied to the determination of MC-LR in water samples.

Albumin coated CuInS2 quantum dots as a near-infrared fluorescent probe for NADH, and their application to an assay for pyruvate

Water-soluble CuInS₂ quantum dots (QDs) stabilized with 3-mercaptopropionic acid were synthesized in aqueous solution and then coated with bovine serum albumin. The resulting particles display fluorescence with a peak at 680 nm that is effectively quenched by 1, 4-dihydro-nicotinamide adenine dinucleotide (NADH), but not by 1, 4-nicotinamide adenine dinucleotide (NAD⁺). The enzyme lactate dehydrogenase catalyzes the reduction of pyruvate and dehydrogenation of lactic acid using NAD⁺ or NADH as a cosubstrate. The new QDs were applied to monitor the course of lactate dehydrogenase-catalyzed reaction of pyruvate by detecting NADH via its quenching effect. This resulted in a convenient and selective detection scheme for pyruvate. The detection limit is as low as 25 nM.

CdS quantum dots as a fluorescent sensing platform for nucleic acid detection

We demonstrate that CdS quantum dots (QDs) can be applied to fluorescence-enhanced detection of nucleic acids in a two-step protocol. In step one, a fluorescently labeled single-stranded DNA probe is adsorbed on the QDs to quench its luminescence. In step two, the hybridization of the probe with its target ssDNA produces a double-stranded DNA which detaches from the QD. This, in turn, leads to the recovery of the fluorescence of the label. The lower detection limit of the assay is as low as 1?nM. The scheme (that was applied to detect a target DNA related to the HIV) is simple and can differentiate between perfectly complementary targets and mismatches.

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.

Biotinylation of quantum dots for application in fluoroimmunoassays with biotin-avidin amplification

A competitive microplate fluoroimmunoassay was developed for the determination of human serum albumin in urine. It is based on the use of biotinylated CdTe quantum dots (QDs) whose synthesis is optimised in terms of storage stability, purification, and signal-to-noise ratio. The bioconjugated QDs were characterised by gel chromatography and gel electrophoresis. Storage stability and quantum yield were investigated. The excitation/emission wavelengths are 360/620?nm. The immunoassay of human serum albumin in urine has a working range from 1.7 to 10?μg.mL^?1, and the limit of detection is 1.0?μg.mL^?1.

Functionalized manganese-doped zinc sulfide quantum dot-based fluorescent probe for zinc ion

We report on a simple strategy for the determination of zinc ion by using surface-modified quantum dots. The probe consists of manganese-doped quantum dots made from zinc sulfide and capped N-acetyl-L-cysteine. The particles exhibit bright yellow-orange emission with a peak at 598?nm which can be attributed to the ⁴T₁→⁶A₁ transition of Mn(II). This bright fluorescence is effectively quenched by modifying the sulfur anion which suppresses the radiative recombination process. The emission of the probe can then be restored by adding Zn(II) which causes the formation of a ZnS passivation layer around the QDs. The fluorescence enhancement caused is linear in the 1.25 to 30?μM zinc concentration range, and the limit of detection is 0.67?μM.

Electrochemiluminescent sensing of dopamine using CdTe quantum dots capped with thioglycolic acid and supported with carbon nanotubes

We have synthesized water-dispersible CdTe quantum dots (QDs) capped with thioglycolic acid. Their quantum yield is higher than 54%. A sensitive electrochemiluminescence (ECL) method was established based on the modification of the composite of the QDs, carbon nanotubes and chitosan on indium tin oxide glass. The sensor displays efficient and stable anodic ECL which is quenched by dopamine. A respective sensor was designed that responds to dopamine linearly in the range of 50?pM to 10?nM, and the detection limit is 24?pM. Dopamine was determined with this sensor in spiked cerebro-spinal fluid with average recoveries of 95.7%.

Manganese-doped ZnS quantum dots as a phosphorescent probe for use in the bi-enzymatic determination of organophosphorus pesticides

We present a sensitive and selective method for the determination of organophosphorus pesticides (OPs) based on the inhibition of the enzyme acetylcholinesterase (AChE). It is making use of quantum dots QDs of the type Mn: ZnS that display long-lived phosphorescence emission and act as optical probes for hydrogen peroxide (H₂O₂). In this assay, acetylcholine (ACh) is first hydrolyzed by AChE, and the enzyme choline oxidase (ChOx) further oxidizes choline under the formation of H₂O₂ which quenches the phosphorescence of the QDs. If, however, OPs are added to the solution, the rate of enzymatic hydrolysis by AChE is retarded. This reduces the rate of production of H₂O₂ and results in a reduced quenching efficiency. The slow decay time of the phosphorescence of the QDs also allows time-resolved luminescence intensity to be measured. This can eliminate background fluorescence from the sample and therefore improves analytical accuracy and the signal-to-noise ratio. Under optimized conditions, there is a linear relationship between luminescence intensity and the concentration of paraoxon in the 1 pM to 1 μM range, with an ~0.1 pM limit of detection which is much lower than that of most existing methods. The phosphorescent probe was applied to determine OPs in spiked real samples. Figure

We present a sensitive and selective method for the determination of organophosphorus pesticides (OPs) based on the inhibition of the enzyme acetylcholinesterase (AChE). It is making use of quantum dots QDs of the type Mn-doped ZnS that display long-lived phosphorescence emission and act as optical probes for hydrogen peroxide (H₂O₂).     

Highly sensitive detection of lead(II) ion using multicolor CdTe quantum dots

Multicolor and water-soluble CdTe quantum dots (QDs) were synthesized with thioglycolic acid (TGA) as stabilizer. These QDs have a good size distribution, display high fluorescence quantum yield, and can be applied to the ultrasensitive detection of Pb(II) ion by virtue of their quenching effect. The size of the QDs exerts a strong effect on sensitivity, and quenching of luminescence is most effective for the smallest particles. The quenching mechanism is discussed. Fairly selective detection was accomplished by utilizing QDs with a diameter of 1.6?nm which resulted in a detection limit of 4.7?nmol?L^?1 concentration of Pb(II). The method was successfully applied to the determination of Pb(II) in spinach and citrus leaves, and the results are in good agreement with those obtained with atomic absorption spectrometry.

Detection of silver(I) ion based on mixed surfactant-adsorbed CdS quantum dots

Mixed cationic and anionic surfactants were adsorbed on cadmium sulfide quantum dots (CdS QDs) capped with mercaptoacetic acid. The CdS QDs can be extracted into acetonitrile with 98 % efficiency in a single step. Phase separation only occurs at a molar ratio of 1:1.5 between cationic and anionic surfactants. The surfactant-adsorbed QDs in acetonitrile solution display stronger and more stable photoluminescence than in water solution. The method was applied for determination of silver(I) ion based on its luminescence enhancement of the QDs. Under the optimum conditions, the relative fluorescence intensity is linearly proportional to the concentration of silver(I) ion in the range between 50 pmol L^?1and 4 μmol L^?1, with a 20 pmol L^?1 detection limit. The relative standard deviation was 1.93 % for 9 replicate measurements of a 0.2 μmol L^?1 solution of Ag(I).

Electrochemiluminescence immunoassay at a nanoporous gold leaf electrode and using CdTe quantun dots as labels

An electrochemiluminescence-based immunoassay using quantum dots (QDs) as labels for the carcinoembryonic antigen (CEA) was developed using an electrode modified with leafs of nanoporous gold. CEA was initially immobilized on the electrode via a sandwich immunoreaction, and then CdTe quantum dots capped with thioglycolic acid were used to label the second antibody. The intensity of the ECL of the QDs reflects the quantity of CEA immobilized on the electrode. Thus, in the presence of dithiopersulfate as the coreactant, the ECL serves as the signal for the determination of CEA. The intensity of the electroluminescence (ECL) of the electrode was about 5.5-fold higher than that obtained with a bare gold electrode. The relation between ECL intensity and CEA concentration is linear in the range from 0.05 to 200?ng.mL^-1, and the detection limit is 0.01?ng.mL^-1. The method has the advantages of high sensitivity, good reproducibility and long-term stability, and paves a new avenue for applying quantum dots in ECL-based bioassays.

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.

CdS/ZnS core-shell quantum dots capped with mercaptoacetic acid as fluorescent probes for Hg(II) ions

We have synthesised water soluble CdS/ZnS core-shell quantum dots (QDs) capped with mercaptoacetic acid (MAA). They were characterised by UV–vis absorption spectroscopy, fluorescence spectroscopy, FT-IR and transmission electron microscopy. Such QDs can be used as fluorescent probes for the determination of metal ions because they quench the fluorescence of the QDs. The QDs exhibit absorption and emission bands at 345?nm and 475?nm respectively, which is more longer wavelength compared to MAA-capped CdS QDs and obviously is the result of the larger particle size. The fluorescence intensity of CdS-based QDs is strongly enhanced by coating them with a shell of ZnS. In addition, such functionalised QDs are more sensitive to Hg(II) ions. Parameters such as pH, temperature and concentration of the QDs have been optimised. A high selectivity and sensitivity toward Hg(II) ions is obtained at pH 7.4 and a concentration of 12.0?mg of QDs per L. Under optimum conditions, the fluorescence intensity of CdS/ZnS QDs is linearly proportional to the concentration of Hg(II) in the range from 2.5 to 280?nM, with a detection limit of 2.2?nM. The effect of potentially interfering cations was examined and confirmed the high selectivity of this material.

Fluorescent turn-on detection of cysteine using a molecularly imprinted polyacrylate linked to allylthiol-capped CdTe quantum dots

CdTe quantum dots capped with thioglycolic acid (TGA) display a strong turn-on fluorescence response if exposed to solutions of cysteine (Cys). In order to exploit this effect, a molecularly imprinted polymer (MIP) for Cys was covalently linked to the QDs via allyl mercaptan. The resulting nanomaterials (QDs, MIP-coated QDs, and nonimprint-coated QDs) were characterized by FTIR and scanning electron microscopy. The adsorption of Cys was studied in phosphate buffer (pH 7.4) with respect to equilibration times (5, 15, and 40 min, respectively), binding constants [2.98, 2.42, and 0.96 (×10⁴ M^?1)], and Langmuir isotherms (R²?=?0.9995, 0.9999, and 0.9983) in the Cys concentration range between 3.33 μM to 500 μM. The method has a detection limit of 0.85 μM (3σ, blank, for n?=?10). The selectivity of the MIP-coated QDs for Cys over 19 other amino acids is similar to that of bare QDs, but MIP-QDs afford better recoveries of Cys from solutions also containing bovine serum albumin (90 %) and fetal bovine serum (97 %), respectively, when compared to the recoveries that are obtained with bare (non-imprinted) QDs (135 % and 120 %). This is probably due to the fact that the outer MIP shell largely reduces protein wrapping, dot aggregation, and matrix inclusion.

Ultrasensitive electrochemiluminescent detection of pentachlorophenol using a multiple amplification strategy based on a hybrid material made from quantum dots,graphene, and carbon nanotubes

We report on a highly sensitive and selective electrochemiluminescence (ECL) based method for the determination of pentachlorophenol (PCP). It is based on a new hybrid material composed of CdS quantum dots (QDs), graphene, and carbon nanotubes (CNTs), and uses peroxodisulfate as the coreactant. The use of this system results in a nearly 18-fold increase in ECL intensity. On interaction between PCP and the QDs, a decrease in ECL intensity is observed at PCP in a concentration as low as 1.0 pM and over a wide linear range (from 1.0 pM to 1.0 nM). The method is hardly affected by other chlorophenols and nitrophenols, and the electrode can be recycled.

Quantum dot induced phototransformation of 2,4-dichlorophenol, and its subsequent chemiluminescence reaction

We have studied the CdTe quantum dot-induced phototransformation of 2,4-dichlorophenol (2,4-DCP) and its subsequent chemiluminescence (CL) reaction. Quantum dots (QDs) of different size and capped with thioglycolic acid were prepared and characterized by molecular spectroscopy, X-ray diffraction and transmission electron microscopy. In the presence of QDs, 2,4-DCP is photochemically transformed into a long-living light emitting precursor which can react with N-bromosuccinimide to produce CL with peak wavelengths at 475 and 550 nm. The formation of singlet oxygen during the phototransformation process was confirmed by the enhancement effect of deuterium oxide on the CL reaction and the change in the UV spectrum of a chemical trap. The CL intensity is linearly related to the concentration of 2,4-DCP in the range from 0.36 to 36 μmol L^?1, and the detection limit (at 3σ) is 0.13 μmol L^?1.