Simple Fluorescence Research Articles

Direct and selective metal-free N6-arylation of adenosine residues for simple fluorescence labeling of DNA and RNA.

We have developed an unprecedented transition metal-free approach for the direct fluorescence turn-on labeling of natural oligonucleotides through selective N6-arylation of adenosine moieties. This method allows the simple and direct fluorescence labeling of natural unmodified DNA and RNA, but is dependent on the secondary structure, favoring single-stranded structures.

Diethylamine fluorescence sensor based on silica hollow sphere photonic crystals.

In this study, based on silica hollow sphere photonic crystals (SHSPCs), a simple and selective fluorescence sensor for high-performance diethylamine detection was developed. The sensor does not involve complicated integration of arrays or tedious synthetic work. During the detection, the SHSPCs could significantly enhance the fluorescence intensity of rhodamine 6G (Rh6G), and meanwhile, adsorb some Rh6G on their surface. Due to the competitive adsorption, diethylamine could free the Rh6G which was adsorbed on the surface of the SHSPCs, thus enhancing the fluorescence emission intensity with the increase of Rh6G concentration. The second enhancement of the fluorescence emission enabled the selective detection of diethylamine. It is remarkable that a simple and readily available dye (Rh6G) facilitates the efficient detection of diethylamine. Moreover, the sensor has good interference immunity, stability and reusability.

Controlling the ligands of CdZnTe quantum dots to design a super simple ratiometric fluorescence nanosensor for silver ion detection.

A super simple ratiometric fluorescence nanosensor has been fabricated by controlling the ligands of CdZnTe quantum dots (QDs), allowing the sensitive and visual detection of silver ions (Ag+). The green-emitting L-cysteine-protected CdZnTe QDs (Lcys-CdZnTe QDs) had a specific response to Ag+ and were used as the reporting probe, while the red-emitting N-acetyl-L-cysteine-protected CdZnTe QDs (NAC-CdZnTe QDs) showed no obvious response to all tested metal ions and were selected as the reference probe. Simply mixing them without any encapsulated synthesis ultimately produced a time-saving, low-cost detection method, allowing the sensitive and visual detection of Ag+ in samples. The proposed nanosensor exhibited a linear range of 0.5-4.0 μM along with a detection limit of 0.17 μM, and has been successfully applied in real tap water and lake water samples. This nanosensor also showed obvious color changes in the detection process and has potential in visual semi-quantitative detection. Our approach may provide a general and feasible strategy for designing ratiometric fluorescence nanosensors, which will attract a wide range of interest in sensing-related fields.

Magnetic-enhanced fluorescence sensing of tumor miRNA by combination of MNPs@PDA with duplex specific nuclease.

Highly sensitive and specific detection of miRNA still remains challenging. In this work, a simple and sensitive fluorescence biosensor has been developed for detection of miRNA by combining the magnetic nanoparticles coated with poly-dopamine (MNPs@PDA) with duplex specific nuclease (DSN). The MNPs@PDA could be easily synthesized via autoxidation of dopamine on the surface of magnetic nanoparticles. The MNPs@PDA could specifically bind with the FAM-labeled single-strand DNA (ssDNA) probes via polyvalent metal-mediated coordination, resulting in the quench of fluorescence signal. The MNPs@PDA exhibited good anti-interference performance in complex matrix. The inner filter effect (IFE) of the MNPs@PDA could be eliminated via magnetic separation. In the presence of specific miRNA, DSN digested the DNA in the DNA–miRNA duplexes and small fragments were formed. The force between these small fragments and MNPs@PDA was negligible, resulting in the occurrence of fluorescence signal. Due to the incorporation of DSN, signal amplification was realized via the recycling process. The established method achieved a low detection limit down to 0.42 pM. The linear concentration range was from 5 pM to 5 nM. Moreover, this method also had high specificity. Remarkably, the target miRNAs extracted from human cells were detected by using the sensing platform.

Tetracycline antibiotics and NH4+ detection by Zn-organic framework fluorescent probe.

A fluorescent probe based on single metal-organic framework material without additional fluorophores and active sites can significantly improve the stability of the probe for detection, and has very important application value in environmental analysis and detection. In this paper, a simple and rapid fluorescence detection method was established with Zn-MOF, which realized the highly sensitive detection of tetracycline antibiotics and NH4+ in water. The prepared Zn-MOF has abundant pores and can exist stably in water. When tetracycline antibiotics are present in Zn-MOF aqueous solution, based on the unique coordination ability between Zn and N, tetracycline antibiotics rich in N will be adsorbed into the pore canals of MOF, and aggregation-induced luminescence will occur. The original non-fluorescent Zn-MOF will immediately produce yellow fluorescence, realizing the detection of tetracycline antibiotics in water, with the limit of detection reaching 0.017 μM in a linear range of 0.02-13 μM. Zn-MOF is further used for the detection of tetracycline antibiotics in actual samples of milk and honey. Oxytetracycline (OTC) with the best fluorescence response of tetracycline antibiotics was coated on Zn-MOF to synthesize OTC@Zn-MOF fluorescent probe. NH4+ will replace the original ligand of Zn-MOF, which will disintegrate MOF and release OTC, resulting in a fluorescence decrease. Therefore, NH4+ can be detected with low limit of detection (0.038 μM) in a linear range of 0 to 3 mM. The probe is expected to be able to detect ammonia in the environment.

A carbon dot doped lanthanide coordination polymer nanocomposite as the ratiometric fluorescent probe for the sensitive detection of alkaline phosphatase activity

The development of sensitive methods for alkaline phosphatase (ALP) activity analysis is an important analytical topic. Based on the stimulus-responsive lanthanide coordination polymer, a simple ratiometric fluorescence sensing strategy was proposed to detect ALP activity. A carbon dot (CD) doped fluorescent supramolecular lanthanide coordination polymer (CDs@Tb-GMP) was prepared with Tb3+ and the ligand guanine single nucleotide (GMP). To construct a ratiometric fluorescence biosensor, the fluorescence of Tb-GMP was used as a response signal, and the fluorescence of CDs was used as a reference signal due to its good stability. When excited at 290 nm, the polymer network Tb-GMP emits characteristic fluorescence at 545 nm, while the CDs encapsulated in the polymer network emit fluorescence at 370 nm. After adding ALP to the system, the substrate GMP can be hydrolyzed by ALP, resulting in the destruction of the polymer network. Accordingly, the fluorescence of Tb-GMP significantly decreased, while the fluorescence of CDs slightly increased due to their release from the polymer network. By comparing the relationship between the fluorescence intensity ratio of the two signals and the concentration of ALP, sensitive detection of ALP could be achieved with the linear range from 0.5 to 80 U L-1 and a detection limit of 0.13 U L-1. Furthermore, the proposed ratiometric sensing system was applied to the detection of ALP in human serum samples with desirable results, indicating potential application in clinical diagnosis.

Ultrasensitive fluorescence detection of antitumor drug methotrexate based on a terbium-doped silica dendritic probe.

A novel strategy was developed for the detection of methotrexate (MTX) via the quenching effect of MTX on the fluorescence intensity of terbium-doped dendritic silica particles (Tb@KCC-1). The fluorescence intensity of Tb@KCC-1 can be effectively quenched by MTX at 546 nm under an excitation wavelength of 233 nm. The quenched fluorescence is proportional to the amount of MTX in both plasma and exhaled breath condensate (EBC) samples. Under the optimal conditions, the linear dynamic ranges of the developed method were 44 nM to 2.2 μM for EBC, 44 nM to 0.22 μM and 0.22-2.2 μM for plasma samples. The limit of detection (LOD) and limit of quantification (LOQ) in both plasma and EBC media are 35 and 116 nM, respectively. The developed method has the benefits of fast analysis time, simple approach, high specificity, and sensitivity for the detection of MTX in both media. This nanoprobe has been successfully utilized for the quantification of MTX in patients' plasma and spiked EBC samples, proving the applicability of the nanoprobe for MTX detection in real samples.

Detecting Pesticide Dodine by Displacement of Fluorescent Acridine from Cucurbit[10]uril Macrocycle.

According to a simple guest-replacement fluorescence turn-on mechanism, we constructed a fluorescent probe system based on cucurbit[10]uril (Q[10]) and protonated acridine (AD) to detect the pesticide dodine (DD). Formation of a homoternary inclusion complex AD2@Q[10] in both aqueous solution and solid state was studied by means of 1H NMR spectroscopy and X-ray crystallography. Although AD can emit strong fluorescence in aqueous solution, the homoternary inclusion complex AD2@Q[10] does not exhibit any fluorescence. Upon the addition of the pesticide DD into the aqueous solution of AD2@Q[10], the AD molecules in the Q[10] cavity are displaced by the pesticide DD, and strong fluorescence recovers. The fluorescent probe system based on Q[10] and AD provided a wide determination of DD from 0 to 4.0 × 10-5 mol·L-1 with a low limit of detection of 1.827 × 10-6 mol·L-1. The guest-replacement fluorescence turn-on mechanism is also confirmed by 1H NMR spectroscopy. Further, the fluorescent probe can directly detect DD residues in real agricultural products, and obvious fluorescence signal was observed under UV irradiation.

New Fluorescence Quenching Approach for Determination of Valsartan in Certain Tablets and Spiked Biological Fluids

A new, simple, selective, sensitive, fast, economical, and reliable fluorescence quenching method for the quantitation of valsartan was investigated using basic fuchsin act as a fluorometric dye. The method was depended on the detection quenching influence of valsartan on the fluorescence intensity of basic fuchsin and the reaction between valsartan and basic fuchsin in a McIlvaine buffer medium at pH = 6 to yield a new basic fuchsin–valsartan non-fluorescent complex. The excitation and emission of basic fuchsin fluorescence signal were identified at 535 and 728 nm, respectively. A fluorescence quenching value (ΔF) displayed a very good linear relationship (R2 = 0.9992) with valsartan concentration ranging from 0.003 to 3 μg/mL, a detection limit as low as 0.0009 μg/mL with a high precision and accuracy (RSD% <3). Significantly, no interference effect was found due to the presence of other ingredients commonly found in medical formulations. The acquired data were statistically compared with those acquired from reported chromatographic method and were observed to be in excellent agreement at a 95% confidence level; the planned fluorescence quenching procedure was subsequently utilized to detected the concentration of valsartan in spiked biological fluids and commercial medical tablets.

Selective recognition of tryptophan by a methylpillar[5]arene-based supramolecular fuorescent probe

Herein we present a simple fluorescence quenching method to selectively recognise and determine L-tryptophan (L-Trp) out of other 19 natural amino acids. Methylpillar[5]arene (MeP5), which is employed as a macrocyclic fluorescent probe, exhibits fluorescence activity in the solution of poor solvents because of aggregation-induced emission (AIE) effect. Fluorescence quenching of MeP5 in the solution of EtOH/CH2Cl2 (98/2, v/v) was observed upon the addition of L-Trp whereas other 19 natural amino acids did not bring about obvious change in fluorescence intensity. 1H NMR titration, fluorescence spectroscopy, mass spectrometry and theoretical analysis revealed that L-Trp can be encapsulated into the cavity of MeP5 to form a stable 1:1 host–guest inclusion complex which accounts for the quenching characteristics. The proposed procedure in this investigation offers an attractive and promising method for the selective detection of L-Trp in a mixture of natural amino acids.

A novel sensitive aptamer-based nanosensor using rGQDs and MWCNTs for rapid detection of diazinon pesticide

An optical apta-nanosensor was designed and developed based on reduced graphene quantum dots (rGQDs) and multi-walled carbon nanotubes (MWCNTs) and applied for the selective detection of diazinon as one of the most widely used organophosphorus pesticides. Considering the GQDs and rGQDs high fluorescence emission and optical stability, efficient optical transducers could be designed and precise detection methods could be developed based on such transducers. Herein, by using rGQDs, diazinon specific aptamer, and MWCNTs, a simple economical fluorescence method has been introduced to detect and measure diazinon with the detection limit of 0.4 nM (0.1 µg/L) in the range of 4–31 nM, meeting the specifications and standards set by the European Union and World Health Organization. To assess apta-nanosensor performance, the real application of the novel apta-nanosensor was then validated by detecting and quantifying diazinon in real samples, including tap water, urine, river water, and agricultural runoff water. The obtained results revealed that, due to the lack of binding affinity for the diazinon specific aptamer, the emission recoveries were kept intact in the presence of other pesticides and thereby, the designed apta-nanosensor was able to promptly detect diazinon pesticide with high accuracy and selectivity in real samples. Accordingly, the accurate apta-nanosensor developed in this study provides a sensitive, inexpensive, fast, and portable device for selective detection of diazinon over other pesticides in contaminated regions. The introduced innovative method could also be applied for other pesticides, providing a practical on-site detection approach for real samples of multi pesticides.

Simple Colorimetric and Fluorescence Chemosensing Probe for Selective Detection of Sn2+ Ions in an Aqueous Solution: Evaluation of the Novel Sensing Mechanism and Its Bioimaging Applications.

An easily accessible colorimetric and fluorescence probe 4-((3-chloro-1,4-dioxo-1,4-dihydronaphthalen-2-yl)amino)benzenesulfonamide (4CBS) was successfully developed for the selective and sensitive detection of Sn2+ in an aqueous solution. The sensing mechanism involves reduction of -C═O into -C-OH groups in 4CBS upon the addition of Sn2+, which initiates the fluorescence turn-on mode. A better linear relationship was achieved between fluorescence intensity and Sn2+ concentration in the range of 0-62.5 μM, with a detection limit (LOD) of 0.115 μM. The binding mechanism of 4CBS for Sn2+ was confirmed by Fourier transform infrared analysis, NMR titrations, and mass (electrospray ionization) spectral analysis. Likewise, the proposed sensing mechanism was supported by quantum chemical calculations. Moreover, bioimaging studies demonstrated that the chemosensing probe 4CBS is an effective fluorescent marker for the detection of Sn2+ in living cells and zebrafish. Significantly, 4CBS was able to discriminate between Sn2+ in human cancer cells and Sn2+ in normal live cells.

Molecular crowding-modulated fluorescence emission of gold nanoclusters: Ligand-dependent behaviors and application in improved biosensing

In this work, we found that the fluorescence emission and sensing performance of gold nanoclusters (GNCs) could be modulated greatly under molecular crowding environment in a ligand-dependent manner. The fluorescence emission of glutathione-protected GNCs (GSH-GNCs) can be significantly improved by molecular crowding while that of bovine serum albumin (BSA)-capped GNCs is notably weakened. Further, it was observed that molecular crowding could facilitate the interaction between GSH-GNCs and protamine, thus triggering a synergetic aggregation-induced enhanced emission (AIEE) effect. The AIEE could be suppressed by subsequent addition of heparin, allowing the development of a simple and efficient fluorescence approach for determination of protamine and heparin. The proposed method presents detection ranges of 0.7–3.0 and 0.05–2.8 μg mL−1 for protamine and heparin with the detection limits of 7.5 and 1.7 ng mL-1, respectively. This work is the first study of how molecular crowding impacts the fluorescence emission of GNCs and their interaction with biomolecules. The results indicate that the modulated properties of GNCs in crowding condition may open new prospects for the development of GNCs-based sensing methods.

Development of a fluorescence aptasensor for rapid and sensitive detection of Listeria monocytogenes in food

In this study, a simple fluorescence aptasensor was developed for rapid and sensitive detection of Listeria monocytogenes in food. The aptasensor was comprised of two parts. Aptamer-functionalized upconversion nanoparticles (UCNPs) recognized and adhered to the L. monocytogenes cells and provided strong fluorescent signals. Aptamer-functionalized magnetic nanoparticles (MNPs), which also recognized and adhered to the cells, were used to concentrate the aptamer-UCNP/L. monocytogenes/aptamer-MNP complex. The fluorescence intensity was measured and calibrated to a concentration series of L. monocytogenes. A low limit of detection of 8 cfu mL−1 was estimated from the range of 68 to 68 × 106 cfu mL−1. Compared with other methods currently available, the proposed method had wider detection range and higher sensitivity. Furthermore, the selectivity and functionality when used to detect L. monocytogenes in real food samples suggest it may be useful for practical applications.

A Heparin based dual ratiometric sensor for Thrombin

Thrombin is an important enzyme that plays a pivotal role in the blood clotting pathways. An imbalance in the activity of this enzyme is clinically known to be associated with various diseases, such as thrombosis, inflammation, atherosclerosis, and haemophilia, suggesting the need to devise sensors for Thrombin detection. However, the majority of the fluorescence-based Thrombin assays rely on fluorescence labelling assays or Thrombin specific recognition biomolecules, such as, aptamers or antibody which requires sophisticated techniques and makes it very expensive. Herein, we report a simple, selective, sensitive and label-free fluorescence detection scheme for Thrombin which is based on the interaction between Thrombin and a fluorescent complex of Heparin with a molecular rotor dye, Thioflavin-T. The detection scheme exploits selective interaction between cationic Thrombin and anionic Heparin to modulate the monomer-aggregate equilibrium of the Thioflavin-T–Heparin system. Importantly, the present system offers a ratiometric response that has the ability for robust quantification of Thrombin concentration even in complex medium. The involvement of all commercially available components is a crucial advantage of this detection scheme. Further, the detection scheme also shows reasonable response in diluted serum matrix.

Fluorescence properties of retinoid X receptor antagonist NEt-SB

Ligands of retinoid X receptors (RXRs) are effective against various diseases, so there is a need for efficient screening methods to discover new ligands. Existing screening methods are complex and time-consuming, and a simple fluorescence assay would be highly desirable. Here, we focused on NEt-SB (4), which has a stilbene structure, as a candidate for this purpose, and examined its fluorescence properties in detail. The fluorescence intensity of 4 was remarkably increased in highly viscous solvents and upon binding to hRXRα-LBD, due to suppression of free rotation of the stilbene moiety. Although the relatively low fluorescence intensity and the short fluorescence wavelength of 4 make this compound itself unsuitable for use in RXR binding assay, our findings provide a basis for further structural evolution, which may lead to a derivative that would be suitable for fluorescence assay of RXR binders.

Detection of water content in alcohol solvents over Al3+ induced colorimetric and NIR-fluorescent sensor based on isophorone-phenylamine

Environment-sensitive fluorescent dyes become universal tools in fluorescence detection and imaging. They are particularly useful as polarity-responsive sensors without destruction. The present study reports a novel isophorone and phenylamine dye (IPA), its recognition ability for Al3+ and its solvatofluorchromic properties in several organic solvents. Structures of IPA was characterized using 1H NMR, 13C NMR, FT-ITR, TOF-MS spectra. Photophysical properties of IPA and its solvatochromic and solvatofluorchromic characteristics were examined using UV–vis and fluorescence spectroscopy. IPA displayed a remarkable “turn-on” NIR emissive and colorimetric response towards Al3+. A correlation between solvent polarity parameter dependent Stokes shifts was interpreted using the Lippert-Mataga model. The results were supported by theoretical calculations. Water content of commercial alcoholic solutions was successfully determined by IPA-Al3+ complex. Therefore, a highly selective and simple fluorescence analysis for Al3+ and trace water in alcohols or alcoholic mixtures was established on basis of intramolecular charge transfer mechanism.

Water-soluble luminescent copper nanoclusters as a fluorescent quenching probe for the detection of rutin and quercetin based on the inner filter effect.

In the present study, we proposed a simple, sensitive and selective fluorescence method for the detection of rutin (Rut) and quercetin (Que) based on the inner filter effect (IFE) utilizing water- soluble cysteine-stabilized copper nanoclusters (Cys-CuNCs) as a fluorescent probe. The Cys-CuNCs were successfully synthesized and characterized using UV-visible absorption, emission, Fourier-transform infrared (FT-IR) spectroscopy, fluorescence lifetime, high resolution transmission electron microscopy (HR-TEM) and zeta potential measurements. Cys-CuNCs exhibited bluish-green luminescence under UV light with characteristic emission maxima at 486 nm. Cys-CuNCs was successfully exploited as fluorescent probes for the detection of Rut/Que. The addition of increasing concentrations of Rut/Que led to a gradual decrease in the emission intensity of Cys-CuNCs. The decrease in Cys-CuNC emission intensities were attributed to the strong IFE and static quenching mechanism. The calculated limit of detection values for Rut and Que were as low as 0.021 μM and 0.035 μM, respectively. The Cys-CuNCs sensing probe exhibited excellent selectivity in the presence of other potential interfering compounds. Furthermore, the present method was successfully applied to the analysis of both Rut and Que in biological samples.

Development of Cu(II)/Cu(I)-induced quantum dot-mediated fluorescence immunoassay for the sensitive determination of ethyl carbamate.

A series of haptens were rationally designed for producing monoclonal antibodies specific for EC and a simple fluorescence immunoassay platform was developed for the sensitive determination of EC based on alkaline phosphatase (ALP)-triggered Cu+ quenching of CdSe quantum dots (QDs). It was noted that Cd as a fluorescence substrate in CdSe QDs can be selectively substituted by Cu+ that resulted in a more significant fluorescence quenching in comparison with Cu2+. Meanwhile, because ALP catalyzed ascorbic acid phosphate and then assisted the transformation of Cu2+ to Cu+, the change in fluorescence intensity was found to be proportional to ALP concentration. After simple magnetic separation, the sensitivity and linear range of the established assay were improved approximately 53-fold and an order of magnitude, respectively, when compared with the conventional ELISA. The proposed platform was able to both amplify the signal and eliminate matrix interferences, making it a promising to determine EC as well as other contaminants in complex food matrix in a highly sensitive and simple manner. Graphical abstract.

Limitation-induced fluorescence enhancement of carbon nanoparticles and their application for glucose detection.

Rational design of detection strategy is the key to high-performance fluorescence analysis. In this article, we found that the glucose-induced limitations can greatly enhance the fluorescence of functionalized carbon nanoparticles (CNPs), which are synthesized through one-step thermal pyrolysis method using phenylboronic acid derivative as the precursors. The glucose can assembly onto the surface of the CNPs to form a "shell", limiting the surfaces' intramolecular rotation and reducing non-radiative decay, which hence resulted in enhanced fluorescence of the CNPs. Under optimal conditions, the fluorescence intensity of the CNPs is nearly 70-fold enhanced, and the method has low detection limit (10μM) and linear response in the concentration range from 50μM to 2000μM. Based on this interesting "target-triggered limitation-induced fluorescence enhancement" phenomenon, a simple and effective non-enzymatic fluorescence enhancement method was developed and successfully applied to the determination of glucose in spiked serum samples. This work provides new insight into the design of fluorescence-enhanced detection strategies based on the limitation-induced property.