We describe the preparation of carbon quantum dots (C-dots) by a one-step hydrothermal method starting from o-aminophenol as the precursor. The C-dots exhibit bright both blue fluorescence (with excitation/emission peaks at 300/410 nm and with quantum yield of 0.40) and green fluorescence (420/500 nm; QY 0.28) without any other element doping. The unique emission properties are attributed to a synergistic effect of amino and hydroxy groups on the surface of the C-dots. The C-dots are shown to be viable fluorescent probes for heparin. The positively charged surface amino groups are assumed to interact with sulfate and carboxy groups in heparin via electrostatic interactions and hydrogen bonding. This causes the blue fluorescence of C-dots to be turned off (quenched). Fluorescence is strongest at a pH value of 6. The fluorometric calibration plot is linear in the 10 to 100 nM concentration range, with an 8.2 nM detection limit (at a signal-to-noise ratio of 3).
Graphical abstract Carbon quantum dots with dual fluorescence emission bands were synthesized and are shown to be a viable fluorescent probe for heparin.
Carbon quantum dots doped with nitrogen and phosphorus were prepared from adenosine 5′-monophosphate (AMP) in a single simple synthesis step. The nitrogen and phosphorus doped C-dots (N,P-C-dots) were characterized by transmission electron microscopy, Fourier transform infrared spectroscopy, fluorescence spectroscopy, X-ray photoelectron spectroscopy and X-ray powder diffraction. These carbon dots display blue fluorescence, with excitation/emission maxima at 360/430 nm, a quantum yield of 26.5% and an average decay time of 4.3 ns. Fluorescence is strongest at neutral pH values but quenched at very high and very low pH values. It is also quenched by ferric ions which suggests the use of the N,P-C-dots as fluorescent probes for Fe(III). A hemolysis test inferred favorable blood compatibility. The fluorescence of the doped C-dots is excitation wavelength dependent and also is susceptible to 2-photon excitation. The nanoparticles were applied in the fluorescent multicolor bioimaging of A549 (adenocarcinomic alveolar basal epithelial) cells under different excitation wavelengths, typically at 405, 488 and 543 nm. Emission colors ranging from blue to green and red can be adjusted in this way.
Graphical abstract Nitrogen and phosphorus doped carbon dots were synthesized and showed excitation wavelength-dependent behavior. They were applied to multi-color fluorescence imaging of adenocarcinomic alveolar basal epithelial cells.
The authors describe a method for the fabrication of a nanohybrid composed of carbon dots (C-dots) and gold nanoparticles (AuNPs) by in-situ reduction of C-dots and hydroauric acid under alkaline conditions. The process does not require the presence of surfactant, stabilizing agent, or reducing agent. The hybrid material was deposited in a glassy carbon electrode (GCE), and the modified GCE exhibited good electrocatalytic activity toward the oxidation of nitrite due to the synergistic effects between carbon dots and AuNPs. The findings were used to develop an amperometric sensor for nitrite. The sensor shows a linear response in the concentration range from 0.1 μmol?L-1 to 2 mmol?L-1 and a low detection limit of 0.06 μmol?L-1 at the signal-to-noise ratio of 3.
Graphical abstract Fabrication, characterization and electrochemical behavior of a glassy carbon electrode modifid with carbon dots and gold nanoparticles for sensing nitrite in lake water.
A colorimetric and fluorescent pH probe was designed by doping carbon dots (C-dots) with Eu(III), Tb(III) and 2,6-pyridinedicarboxylic acid (DPA). The resulting nanoparticles were applied as fluorescent indicators for pH values (best detected at excitation/emission wavelengths of 272/545, 614 nm). The pH induced optical effects are due to pH induced variations in energy transfer. The fluorescence of the probe shows a continuous color variation, and a linear change with pH values in the range from 3.0 to 10.0 can be established by using a Commission Internationale de L’Eclairage (CIE) chromaticity diagram. This new kind of pH nanoprobe is more accurate than previously reported pH indicator probes because the pH value can be calculated by using chromaticity coordinates that only depend on the chromaticity. The pH nanoprobe was applied to visualize pH values in human breast adenocarcinoma cells (MCF-7).
Graphical abstract Carbon dots modified with Eu(III) and Tb(III) complexes of 2,6-pyridinedicarboxylic acid (DPA) were prepared. The doped carbon dots were used as a pH-sensitive nanosensor. The fluorescence chromaticity of the nanoparticles changes with the variation of pH value.
This paper describes an electrochemiluminescent (ECL) based method for chiral recognition and detection of both glutamate (Glu) enantiomers. The luminophore luminol (Lum) was used as both the reductant and stabilizer of Ag nanoparticles (AgNP-Lum) which were combined with carbon quantum dots (C-dots) and placed on a glassy carbon electrode (GCE) along with the enzyme glutamate oxidase (GluOx). The use of these materials is found to result in strong amplification of ECL. The nanomaterials used were characterized by transmission electron microscopy (TEM) and Fourier transform infrared (FTIR). The stepwise fabrication of the electrode was verified by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Under optimized conditions and by applying a typical potential of 0.6 V, the ECL increases linearly in the 5.0 μM to 5.0 mM Glu concentration range, with a 1.6 μM lower detection limit and satisfactory selectivity. A Glu logic gate idea has been designed that is based on this enzymatic biosensor.
Graphical abstract Schematic presentation of the electrochemiluminescent (ECL) biosensor. A glassy carbon electrode (GCE) was modified with C-dots and silver nanoparticles which were deoxidized by luminol (AgNP-Lum) for enzymatic specific detection.
Gold nanoclusters (AuNCs) protected with a bovine serum albumin (BSA) coating are known to emit red fluorescence (peaking at 650 nm) on photoexcitation with ultraviolet light (365 nm). On addition of Cu(II) ions, fluorescence is quenched because Cu(II) complexes certain amino acid units in the BSA chain. Fluorescence is, however, restored if pyrophosphate (PPi) is added because it will chelate Cu(II) and remove it from the BSA coating on the AuNCs. Because PPi is involved in the function of telomerase, the BSA@AuNCs loaded with Cu(II) can act as a fluorescent probe for determination of the activity of telomerase. A fluorescent assay was worked out for telomerase that is highly sensitive and has a wide linear range (10 nU to 10 fM per mL). The fluorescent probe was applied to the determination of telomerase activity in cervix carcinoma cells via imaging. It is shown that tumor cells can be well distinguished from normal cells by monitoring the differences in intracellular telomerase activity.
Graphical abstract Gold nanoclusters (AuNCs) protected by bovine serum albumin (BSA) and displaying red photoluminescence were prepared as fluorescent probe for the determination of telomerase activity and used for imaging of cervix carcinoma (HeLa) cells.
The authors describe three fluorescein-conjugated peptides generated by cell-phage display for use as a diagnostic probes for fluorescent detection and imaging of Salmonella enteritidis and Salmonella typhimurium. The authors also designed a polyvalent-directed peptide polymer synthesized with poly-D-lysine and bifunctional succinimidyl 3-(2-pyridyldithio)propionate with an affinity and sensitivity that is higher by more than an order of magnitude compared to single peptides due to multiple binding site interactions. In order to establish a diagnostic system for food poisoning, imaging analysis was performed using fluorescence microscopy. The limit of detection of the diagnostic system based on polyvalent directed peptide interaction is 102 colony-forming units per mL for Salmonella.
Graphical abstract Schematic of a fluorescent method for detection and imaging of Salmonella enteritidis and Salmonella typhimurium by using a fluorescein labeled polyvalent-directed peptide polymer (PDPP) with a high affinity and sensitivity as a diagnostic probe. The system uses a microplate reader and was applied to the detection of food poisoning.
The authors describe new bifunctional mesoporous silica nanoparticles (NPs) for specific targeting of tumor cells and for intracellular delivery of the cancer drug doxorubicin (DOX). Mesoporous silica nanoparticles (MSNPs) were coated with blue fluorescent N-graphene quantum dots, loaded with the drug DOX, and finally coated with hyaluronic acid (HA). Cellular uptake of the NPs with an architecture of the type HA-DOX-GQD@MSNPs enabled imaging of human cervical carcinoma (HeLa) cells via fluorescence microscopy. The cytotoxicity of the nanoparticles on HeLa cells was also assessed. The results suggest that the NPs are higher cytotoxicity effect and exert in living cell imaging ability. Compared to the majority of other drug nanocarrier systems, the one described here enables simultaneous DOX release and fluorescent monitoring.
Graphical abstract Schematic of the bifunctional mesoporous silica nanoparticles were obtained via the Stöber method, along with the doxorubicin loaded and the hyaluronic acid capped. The sensor shows good specificity and significant cytotoxicity effect on Hela cells. (TEOS: tetraethyl orthosilicate; GQDs: graphene quantum dots; DOX: doxorubicin; HA: Hyaluronic acid).
Carbon polymer dots (CPDs) were prepared by a one-pot aqueous synthetic route from ascorbic acid and diethylenetriamine at room-temperature. The CPDs under 350-nm excitation exhibit blue fluorescence peaking at 430 nm with a quantum yield of 47%. Other features include an average diameter of 5 nm, a fluorescence that is independent of the excitation wavelength, good water dispersibility and photostability, and excellent biocompatibility. The CPDs are shown to be viable fluorescent probes for ferric ion which acts as a strong quencher. The response to Fe(III) is linear in the 0.2 to 10 μM concentration range, and the detection limit is 0.1 μM. The probe was applied to the determination of Fe(III) in environmental waters and to intracellular imaging of ferric ions in HeLa cells.
Graphical abstract Carbon polymer dots (CPDs) are prepared from ascorbic acid and diethylenetriamine (DETA) at room-temperature (RT). The RT-CPDs exhibit excellent optical performance, biocompatibility and selectivity of quenching by ferric ions. This can be applied for determination and intracellular imaging of ferric ion.
A multiplexed graphene oxide (GO) fluorescent nanoprobe is described for quantification and imaging of messenger RNAs (mRNAs) in living cells. The recognizing oligonucleotides (with sequences complementary to those of target mRNAs) were labeled with different fluorescent dyes. If adsorbed on GO, the fluorescence of the recognizing oligonucleotides is quenched. After having penetrated living cells, the oligonucleotides bind to target mRNAs and dissociate from GO. This leads to the recovery of fluorescence. Using different fluorescent dyes, various intracellular mRNAs can be simultaneously imaged and quantified by a high content analysis within a short period of time. Actin mRNA acts as the internal control. This GO-based nanoprobe allows mRNA mimics to be determined within an analytical range from 1 to 400 nM and a detection limit as low as 0.26 nM. Up to 3 intracellular mRNAs (C-myc, TK1, and actin) can be detected simultaneously in a single living cell. Hence, this nanoprobe enables specific distinction of intracellular mRNA expression levels in cancerous and normal cells. It can be potentially applied as a tool for detection of cancer progression and diagnosis.
Graphical abstract A multiplexed graphene oxide (GO)-based fluorescent nanoprobe is described for quantification and imaging of intracellular messenger RNAs. After penetrating living cells, the recovered fluorescence of the dissociated recognizing oligonucleotides can be analyzed , and this allows for simultaneous detection of up to 3 intracellular messenger RNAs.
Carbon dots (CDs) possess superior fluorescent properties in that they do not blink, are biocompatible, chemically inert, have small size and well tunable photoluminescence (PL), can be easily functionalized with biomolecules, and can be multi-photon excited to give up-converted PL. This review (with 141 refs.) summarizes recent progress in the field of imaging using carbon dots doped with heteroatoms (X-CDs). Following an introduction, we discuss top-down and bottom-up strategies for synthesis and methods for surface modification. We also compare the differences in synthesis for undoped CDs and X-CDs. Specifically, CDs doped with heteroelemets nitrogen, phosphorus, sulfur, selenium, boron and silicium are treated. We then discuss method for determination of the properties (particle size, ZP), how doping affects fluorescence (spectra, quantum yields, decay times), and how dopants affect upconversion (UC, anti-Stokes luminescence). We finally review the progress made in fluorescent imaging of cells tissue, and other biomatter. This review also gives new hints on how to use synthetic methods for tuning the structure of X-CDs, how doping affects properties, and how to achieve new bioimaging applications.
Graphical abstract Carbon dots doped with heteroatoms (X-CDs) are a kind of fluorescent nanomaterials that display bright fluorescence, high quantum yield, photostability, biocompatibility and low toxicity. Hence, they possess large potential for both in-vitro and in-vivo bioimaging.
A method is described for the determination of the polarity of mixed organic solvents by using the fluorescent probe Hostasol Red (HR) desposited on the outer surface of nanosized zeolite L. Organic solvents and their mixtures can be roughly classified according to their polarity with bare eyes and fluorometrically. Emission peaks range from 520 to 640 nm. Some solvents act as quenchers. The method is studied with series of protic and nonprotic solvents, and with selected mixtures of organic solvents.
Graphical abstract The dye Hostalene Red adsorbed on nanosized zeolite shows strong fluorescence solvatochromism. This can be exploited to quickly assess the polarity of solvents and solvent mixtures.
The authors describe a method for the differentiation of penicillamine (PA) enantiomers by using CdSe/ZnS quantum dots (QDs) modified with β-cylodextrin (β-CD-CdSe/ZnS QDs). Selective enantiorecognition of L-PA and D-PA was accomplished by virtue of selective host-guest interaction between the PAs and the β-CD pockets on the QDs. The fluorescence intensity of the modified QDs decreases in the presence of L-PA. On the contrary, it increases in the presence of D-PA. These findings form the basis for a new method for recognition of PA enantiomers. Under optimized conditions, a linear relationship exists between fluorescence intensity and D-PA concentration in the 0.1 to 5.0 mg L?1 range, and between 0.8 and 5.0 mg L?1 for L-PA. Detection limits are 0.06 mg L?1 for D-PA, and 0.2 mg L?1 for L-PA. The potential of this method has been demonstrated by the determination of D-PA in pharmaceutical formulations and L-PA in (spiked) environmental samples.
Graphical abstract Selective and specific enantiorecognition of penicillamine (PA) enantiomers using β-cylodextrin modified CdSe/ZnS quantum dots is described. Fluorescence intensity increases in the presence of D-PA, but it decreases in the presence of L-PA. Results were the basis for analytical applications.
Near infrared (NIR) emitting semiconductor quantum dots can be excellent fluorescent nanoprobes, but the poor biodegradability and potential toxicity limits their application. The authors describe a fluorescent system composed of graphene quantum dots (GQDs) as NIR emitters, and novel MnO2 nanoflowers as the fluorescence quenchers. The system is shown to be an activatable and biodegradable fluorescent nanoprobe for the “turn-on” detection of intracellular glutathione (GSH). The MnO2-GQDs nanoprobe is obtained by adsorbing GQDs onto the surface of MnO2 nanoflowers through electrostatic interaction. This results in the quenching of the NIR fluorescence of the GQDs. In the presence of GSH, the MnO2-GQDs nanoprobe is degraded and releases Mn2+ and free GQDs, respectively. This gives rise to increased fluorescence. The nanoprobe displays high sensitivity to GSH and with a 2.8 μM detection limit. It integrates the advantages of NIR fluorescence and biodegradability, selectivity, biocompatibility and membrane permeability. All this makes it a promising fluorescent nanoprobe for GSH and for cellular imaging of GSH as shown here for the case of MCF-7 cancer cells.
Graphical abstract A biodegradable NIR fluorescence nanoprobe (MnO2-GQDs) for the “turn-on” detection of GSH in living cell was established, with the NIR GQD as the fluorescence reporter and the MnO2 nanoflower as the fluorescence quencher.
A composite consisting of carbon nanotubes (CNT) and copper nanoparticles (CuNPs) was prepared by a chemical reduction method, and its structure characterized by scanning electron microscopy, transmission electron microscopy energy dispersive spectroscopy and FT-IR spectrometry. The hybrid composite was deposited on the surface of a disposable gold electrode that was manufactured from a commercial digital versatile gold disc by a drop casting method. The electrochemical properties of the modified electrode were investigated by cyclic voltammetry and differential pulse voltammetry. The sensor showed an excellent electrocatalytic activity towards oxidation of paracetamol (PA). The calibration plot (with current typically measured at 0.41 V vs. Ag/AgCl) is linear in the 0.5 to 80 μM concentration range, and the detection limit is as low as 10 nM. The sensor was successfully applied to the determination of PA in spiked water and tablet samples where it gave recoveries ranging between 95.25 and 100.5 %.
Graphical abstract Carbon nanotubes (CNT) -copper nanoparticles (CuNPs) hybrid composite was synthesized by a facile method then the nanohybrid was used as a modifier for the DVD gold electrode for improving its performance toward paracetamol electrooxidation. Cyclic voltammetry and differential pulse voltammetry were used for characterization and determination of paracetamol, respectively.
The authors describe a fluorescent probe for sulfide that is based on carboxy-functionalized semiconducting polymer dots (P-dots). The dots were prepared from carboxy-functionalized poly[(9,9-dioctylfluorenyl-2,7-diyl)-co-(1,4-benzo-2,1′-3-thiadiazole)] (referred to as COOH-PFBT) via co-precipitation. The P-dots aggregate on addition of Cu(II) ions and their green fluorescence (with excitation/emission peaks at 455/540 nm) is then quenched. Fluorescence is restored on addition of sulfide to the aggregates due to the formation of CuS. This quenching-recovery (“off-on”) mechanism forms the basis for a new sulfide detection scheme. Fluorescence increases linearly in the 1.25 to 75.0 μM sulfide concentration range, with a 0.45 μM detection limit. Good selectivity over other anions is demonstrated. The method shows recoveries ranging between 98.6% and 105.7% when applied to the determination of sulfide in spiked real water samples.
Graphical abstract Schematic of a fluorescent off-on sensor for (hydrogen) sulfide based on the use of semiconducting polymer dots (PFBT-COOH) whose fluorescene is quenched by Cu(II) ion but restored on addition of (hydrogen) sulfide.
An ultrasensitive paper based lateral flow assay is described for rapid and simultaneous fluorometric detection of several β-agonists including clenbuterol and its chemical analogues (mabuterol, brombuterol, cimaterol, cimbuterol, bromchlorbuterol and banbuterol). A nonspecific monoclonal antibody (mAb) against clenbuterol and its analogues was prepared and employed in a competitive immunoassay where mAb conjugated to fluorescent nanoparticles and free β-agonists compete for the binding sites. This enables rapid screening for the 7 β-agonists in a single run that takes about 8 min. Detection limits for the seven β-agonists are <50 pg g?1 of pork. Recoveries ranged from 69.5% to 102.4%, and relative standard deviations were ±15%. The assay was applied to the analysis of both using spiked and unspiked pork for β-agonists, and the results compare well to those obtained by HPLC-MS.
Graphical abstract Schematic presentation of an ultra sensitive fluorescent nanoparticle based paper based assay for rapid detection of multi β-agonists in pork tissue.
An electrochemical approach is introduced for synthesis of carbon dots (CDs) by exfoliating graphite rods at a voltage of 15 V in an electrolyte consisting of a mixture of water and two ionic liquids. It is found that the size of the CDs can be tuned by varying the fraction of water in the mixed electrolyte; CDs in sizes of 4.9, 4.1 and 3.1 nm are obtained if the electrolyte contains water in fractions of 24, 38 and 56 %, respectively. The CDs have a quantum yield of almost 10 % and display the typical excitation wavelength-dependent maxima of photoluminescence, strongest at excitation/emission wavelengths of 360/440 nm. Fourier transform infrared and X-ray photoelectron spectroscopy show the CDs to have oxygen functional groups on their surface which strongly improve solubility. The CDs were applied to image cells of the electricity-producing bacteria Shewanellaoneidensis MR-1.
Graphical Abstract An electrochemical approach is introduced to synthesize carbon dots by exfoliating graphite rods in mixed electrolyte of water and ionic liquids. The increasing size of carbon dots was realized by reducing the volume of water in the mixed electrolyte. The carbon dots were used to fluorescently image the electricity-producing bacterium Shewanellaoneidensis MR-1.
We report on a sensitive and selective fluorescent assay utilizing native carbon dots (CDs) as signal transducers. The optical probes 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) or 3,3′-diaminobenzidine (DAB) were employed as substrates of horseradish peroxidase (HRP). It was found that the corresponding oxidation products (ox-ABTS or ox-DAB) quench the fluorescence of CDs, mainly via photoinduced electron transfer in case of ox-ABTS, and via aggregation and inner filter effect in case of ox-DAB. By coupling with bienzyme (glucose oxidase and HRP)-mediated biocatalytic reactions, the method was applied to the determination of hydrogen peroxide and glucose. In case of ABTS as the substrate of HRP, a wide linear range (0.05 to 100 μM) and a very low detection limit (10 nM) for glucose were attained. The method was applied to the determination of glucose in human serum and the results were found to agree well with data provided by a local hospital.
Graphical abstract In this work, a sensitive and selective fluorescent assay was developed for probing the enzymatic substrates of hydrogen peroxide and glucose which utilized native carbon dots (CDs) as signal transducer.
We have prepared graphene quantum dot-europium(III) complex composites by noncovalently connecting chelating ligands dibenzoylmethane (DBM) and 1,10-phenanthroline (Phen) with graphene quantum dots (GQDs) first, followed by coordination to Eu(III). The resulting composites are well water-soluble and display red fluorescence of high color purity. The composites were characterized by transmission electron microscopy, X-ray photoelectron spectroscopy and X-ray diffraction. Aqueous solutions of the composites under 365 nm excitation display fluorescence with a peak at 613 nm and a quantum yield as high as 15.5 %. The good water solubility and stable photoluminescence make the composites very different from other Eu(III)-based coordination complexes. The composites are cell viable and can be used to label both the cell membrane and the cytoplasm of MCF-7 cells. They are also shown to act as bioprobes for in-vivo localization of tumorous tissue. In our perception, such composites are expected to possess wide scope because of the many functionalizations that are possible with GQDs.
