Ligand-Induced Red-Emitting Copper Nanoclusters for Selective Fluorescence Determination of Aluminum Ions.

Monitoring levels of excessive aluminum ions (Al3+) is crucial as it can harm the immune system, reduce enzyme activity, cause cell death, and damage environmental and biological systems. Developing a fast and efficient Al3+ ion determination method is the key to addressing this issue. In this work, red-emitting fluorescent copper nanoclusters (CuNCs) were synthesized using N-acetyl-l-cysteine (NAC) as a ligand and CuCl2·2H2O through a facile procedure. The NAC-CuNCs exhibited a large Stokes shift and displayed remarkable luminescence properties. A method for detecting Al3+ through a fluorescence probe was proposed. Its fluorescence mechanism was also explored. The probe showed rapid responsiveness (within 1 min) to Al3+ ion determination. The detection limit for Al3+ was found to be 19.7 nM, which is significantly lower than the WHO's value and most reports, with a linear range of 0-52.9 µM. The determination of Al3+ concentrations in actual water using the fluorescence probe yielded satisfactory outcomes. Moreover, the visual detection of Al3+ ions was also achieved through a smartphone, which can enhance its fast and practical detection.

Biomass-derived carbon dots as emerging visual platforms for fluorescent sensing.

Biomass-derived carbon dots (CDs) are non-toxic and fluorescently stable, making them suitable for extensive application in fluorescence sensing. The use of cheap and renewable materials not only improves the utilization rate of waste resources, but it is also drawing increasing attention to and interest in the production of biomass-derived CDs. Visual fluorescence detection based on CDs is the focus of current research. This method offers high sensitivity and accuracy and can be used for rapid and accurate determination under complex conditions. This paper describes the biomass precursors of CDs, including plants, animal remains and microorganisms. The factors affecting the use of CDs as fluorescent probes are also discussed, and a brief overview of enhancements made to the preparation process of CDs is provided. In addition, the application prospects and challenges related to biomass-derived CDs are demonstrated.

Nitrogen-doped biomass-derived carbon dots for fluorescence determination of sunset yellow.

Sunset Yellow (SY) is a widely used food coloring in the food industry. However, exceeding the allowable limit of this dye poses a significant threat to human health. To address this issue, we developed Lycium ruthenicum-derived nitrogen-doped carbon dots (N-CDs) with a stable blue fluorescence through hydrothermal treatment for SY determination. The quantum yield (QY) of these N-CDs was found to be up to 10.63%. Physical characterization of N-CDs was performed using various spectroscopic techniques to confirm their excellent photostability and non-toxic properties. Furthermore, the presence of SY had a substantial quenching effect on the fluorescence intensity (F0/F) of the N-CDs. Leveraging this observation, we developed a fluorescent sensor for the determination of SY in the concentration range of 0.05 to 35.0 µM, with a limit of detection (LOD, 3σ/K) of 17 nM. The excellent fluorescent sensor also showed satisfactory results in the practical drink samples. Moreover, the stability and cytotoxicity of N-CDs as a fluorescent probe were studied. Finally, the N-CDs were applied to cell imaging using A549 cells.

Visual Detection and Sensing of Mercury Ions and Glutathione Using Fluorescent Copper Nanoclusters.

Visual detection of mercury ions and glutathione is of great significance to public health and environmental issues. Herein, we developed a fluorescent sensor (l-Cys/CuNCs@ESM) based on the eggshell membrane (ESM) and red-emitting copper nanoclusters (CuNCs) by the in situ strategy via l-cysteine (l-Cys) as the reducing and protective agent for mercury ions and glutathione sensing visually. The as-prepared fluorescent product had good stability, portability, large Stokes shift (250 nm), and long fluorescence lifetime (7.3 µs). Notably, the l-Cys/CuNCs@ESM exhibited a specific fluorescence quenching response toward Hg2+. Moreover, the interaction between glutathione (GSH) and Hg2+ could subsequently recover the fluorescence effectively. Inspired by this "on-off-on" switch, the l-Cys/CuNCs@ESM was applied as the dual-sensing system for visual detection of mercury ions and glutathione integrating with the portable smartphone. The limit of detection (LOD) of Hg2+ is 1.1 µM for visualization and 0.52 µM for the fluorescence spectrometer. The corresponding LODs of GSH are 2.8 and 0.59 µM, respectively. This platform presents significant sensitivity, specificity, and stability, offering a promising potential for real-time/on-site sensing.

An alkaline phosphatase-induced immunosensor for SARS-CoV-2 N protein and cardiac troponin I based on the in situ fluorogenic self-assembly between N-heterocyclic boronic acids and alizarin red S.

Alkaline phosphatase (ALP)-induced in situ fluorescent immunosensor is less investigated and reported. Herein, a high-performance ALP-labeled in situ fluorescent immunoassay platform was constructed. The developed platform was based on a fluorogenic self-assembly reaction between pyridineboronic acid (PyB(OH)2) and alizarin red S (ARS). We first used density functional theory (DFT) to theoretically calculate the changes of Gibbs free energy of the used chemicals before and after the combination and simulated the electrostatic potential on its' surfaces. The free ARS and PyB(OH)2 exist alone, neither emits no fluorescence. However, the ARS/PyB(OH)2 complex emits strong fluorescence, which could be effectively quenched by PPi based on the stronger affinity between PPi and PyB(OH)2 than that of ARS and PyB(OH)2. PyB(OH)2 coordinated with ARS again in the presence of ALP due to the ALP-catalyzed hydrolysis of PPi, and correspondingly, the fluorescence was restored. We chose cTnI and SARS-CoV-2 N protein as the model antigen to construct ALP-induced immunosensor, which exhibited a wide dynamic range of 0-175 ng/mL for cTnI and SARS-CoV-2 N protein with a low limit of detection (LOD) of 0.03 ng/mL and 0.17 ng/mL, respectively. Moreover, the proposed immunosensor was used to evaluate cTnI and SARS-CoV-2 N protein level in serum with satisfactory results. Consequently, the method laid the foundation for developing novel fluorescence-based ALP-labeled ELISA technologies in the early diagnosis of diseases.

Yeast powder derived carbon quantum dots for dopamine detection and living cell imaging.

Dopamine (DA) is an important neurotransmitter in the brain of mammals. There is a critical need for fast and sensitive determination approaches to monitor these potential diseases due to various weaknesses in clinical trials of the existing methods for DA detection. DA can effectively quench the fluorescence of carbon quantum dots (CDs) through the inner filter effect and static quenching. In this work, fluorescent yeast CDs (Y-CDs) were prepared via a simple hydrothermal approach of using yeast powder and regarded as the fluorescent nanoprobe to directly monitor the DA concentration. The as-prepared detection platform exhibited excellent sensitivity and selectivity toward DA with a low detection limit of 30 nM and a wide linear range of 0.05-150 µM. Benefiting from these outstanding features, the developed label-free method has been successfully applied for fast DA detection in human serum samples with satisfactory recoveries. Furthermore, it demonstrated that the Y-CDs were well suitable for live cell imaging and showed low toxicity toward MCF-7 cells. Consequently, this work will facilitate the great potential of the versatile Y-CDs in developing biosensors for clinical diagnosis and other biological applications.

Simple construction of a two-component fluorescent sensor for turn-on detection of Hg2+ in human serum.

The simply constructed fluorescent sensor with inexpensive reagents and low toxicity has attracted increasing attention contributing to its practical application. However, the common construction methods usually required a few building blocks and complex procedures, which is inconvenient for their further application. Herein, a simply constructed fluorescent Hg2+ sensor has been developed based on the intrinsic fluorescence quenching power of G-quadruplex. Two components, AGRO 100 and AMT, were used to construct the sensor. AMT was selected as the fluorescent probe because of its distinct merits. The free AMT emits strongly. However, the fluorescence of AMT could be quenched by G-quadruplex DNA. Additionally, AMT is less toxic and inexpensive. AGRO 100 acts as both the quencher and the capture sequence because it consists of G-rich sequences and T-T mismatched base pairs. The fluorescence of AMT could be quenched by the formed G-quadruplex structure of AGRO 100 in the presence of K+. In the presence of Hg2+, G-quadruplex structure of AGRO 100 was switched to hairpin DNA structure because T-T mismatched base pairs in AGRO 100 could specifically recognize and capture Hg2+ with high affinity. Thus, AMT was released and the fluorescence of AMT was recovered. The developed sensing system was successfully applied to detect Hg2+ in human serum with good recovery and reproducibility.

A colorimetric and ratiometric photometric sequential assay for ascorbic acid and alkaline phosphatase in serum based on valence states modulation.

The photometric method is widely used in real clinical tests due to its simple operation, low cost and convenient. Many of the reported colorimetric ALP assays so far are non- ratiometric because the detection was based on changes in absorbance at a single wavelength. The development of novel colorimetric and ratiometric assay is of importance for quantitatively measuring target with high accuracy. The challenge in the design of ratiometric photometric assay is that the chromophore must have a significant spectral shift before and after binding to the target. Here, we report a colorimetric and ratiometric photometric sequential assay of AA and ALP based on the complexation between ARS and Cu2+ and redox reaction between AA and Cu2+. The absorption band of ARS centered at 425 nm (yellow color), which could be shifted to 510 nm (red color) upon Cu2+ binding. However, as far as we know, this classic color reaction has not been used to develop a ratiometric photometric method to sequentially detect AA and ALP, although photometric methods based on the regulation of other color reagents with oxidizing metal ions have been reported. The proposed sensing system shows a limit of detection for ALP at 0.24 U L-1 and could be applied for detecting ALP in newborn calf serum. The established sensing system makes a useful contribution to the detection of ALP in complex clinical samples.

A comparison of PMT-based and CCD-based sensors for electrochemiluminescence detection of sunset yellow in soft drinks.

The use of artificial colorants in food is highly regulated due to their potential to harm human health. Thus, it is crucial to detect these substances effectively to ensure conformance with industrial standards. In this work, we prepared a photomultiplier tube (PMT)-based electrochemiluminescence (ECL) sensor and a charged coupled device (CCD)-based ECL sensor and compared their merits in the detection of sunset yellow (SY) dye. The sensors used C,N quantum dot-embedded g-C3N4 nanosheets (QDs@NSs) as the ECL agent and K2S2O8 as the coreactant. SY was analyzed on the basis of amplification in the QDs@NHs-K2S2O8 ECL system. The PMT-based sensor realized ultrasensitive detection using a single electrode, especially at low concentrations of SY. A CCD-based sensor imaged the ECL phenomenon of an electrode array and provided the advantages of high throughput and time savings. Under optimized conditions, both sensors exhibited high specificity, reproducibility and stability; detection limits of 20 nM with PMT detection and 5 µM with CCD detection were determined for SY, with detection ranging over at least two decades. The practical feasibilities of these systems were confirmed by satisfactory detection of SY in real drink samples.

Novel synthesis of orange-red emitting copper nanoclusters stabilized by methionine as a fluorescent probe for norfloxacin sensing.

In the present work, we report a novel chemical approach for the synthesis of orange-red emitting copper nanoclusters (Cu NCs) using L-methionine as stabilizing agent at room temperature for the first time. The synthetic route is facile, economical and viable. The methionine stabilized copper nanoclusters (Cu NCs/Met) were thoroughly characterized by TEM, FT-IR, XPS, UV-Vis, steady state and transient fluorescence spectroscopy. The results show the synthesized Cu NCs/Met with a fluorescence quantum yield of 4.37% possessed high stability and excellent optical features such as large Stokes shift and long fluorescence lifetime (8.3 µs). Significantly, the fluorescence intensity of Cu NCs/Met could be efficiently quenched by norfloxacin (NOR) pharmaceutical. A fast and cost-effective NOR sensor was proposed employing Cu NCs/Met as the fluorescent nanoprobe, and the quenching mechanisms were attributed to inner filter effect and agglomeration-induced quenching. The developed sensor exhibited a high sensitivity and selectivity towards NOR in a wide linear range from 0.05 to 250 µM with a detection limit as low as 17 nM. Moreover, the practicability of the developed NOR sensor for real sample assay was validated with satisfactory recoveries, indicating this sensing platform with great potential for label-free pharmaceutical detection in complex systems.

Design of a Fluorescence Turn-on and Label-free Aptasensor Using the Intrinsic Quenching Power of G-Quadruplex to AMT.

A novel fluorescent aptasensor based on the G-quadruplex induced fluorescent quenching of psoralen and the competitive interactions between 4'-aminomethyl-4,5',8-trimethylpsoralen (AMT), adenosine triphosphate (ATP) and G-rich DNA functionalized split ATP aptamer was proposed. The binding of ATP to the G-rich DNA functionalized split aptamer induced a significant enhancement in fluorescence emission intensity while undergoing excitation at 340 nm. Under the optimal conditions, the developed aptasensor showed high selectivity and good accuracy for detecting ATP. The practicality of the proposed aptasensor has been confirmed by successfully analyzing ATP in spiked human blood serum samples with satisfactory results. As far as we know, this is the first time that the intrinsic quenching ability of G-quadruplex was applied to simply construct a fluorescence turn-on and label-free aptasensor. On account of the superiority of the simplicity of the design strategy, more work is expected in the future to develop a variety of novel sensors for other important analytes using the quenching capability of G-quadruplex through reasonable designs.

In situ fabrication of a luminescent copper nanocluster/eggshell membrane composite and its application in visual detection of Ag+ ions, light-emitting diodes and surface patterning.

In this work, we report a novel strategy for fabricating a luminescent 2D nanocomposite at room temperature by in situ generation of luminescent copper nanoclusters (Cu NCs) embedded in natural monolithic eggshell membrane (ESM) using dithiothreitol as the reducing and capping agent. The established fabrication is facile, cost-effective and viable. The as-prepared Cu NC/ESM nanocomposite exhibited excellent photoluminescence performance, improved chemical, thermal and photo stability, convenient tailoring and flexibility. Significantly, the nanocomposites could be employed as test strips for the visual detection of Ag+ ions based on the luminescence quenching phenomenon and as color conversion layers in light-emitting diodes. Furthermore, application of the proposed strategy for surface luminescence patterning was well demonstrated, indicating great potential in biomass based anti-counterfeiting, information encryption and security paper or sheets.

Novel strategy to improve the sensing performances of split ATP aptamer based fluorescent indicator displacement assay through enhanced molecular recognition.

Split aptamer strategy was often used to improve the sensitivity of aptasensor. However, traditional split aptamer strategy can not be directly used to improve the label-free aptamer based Thioflavin T (ThT) displacement assay for ATP because the split ATP aptamer display much lower enhancement effects on the fluorescence of ThT than intact aptamer. In order to address this issue, this is the first report using G-rich DNA sequence to enhance the affinity of the two split ATP aptamer halves to ThT and offer lower limit of detection (LOD), wider linear range and higher selectivity through the enhanced molecular recognition. Compared to the intact aptamer/ThT complex, the ensemble of two G-rich split ATP aptamer fragments/ThT are higher fluorescent. Consequently, G-rich sequences would improve the fluorescent signal and thus the sensing performance of the proposed assay. In the optimized conditions, the LOD of the proposed fluorescent ATP aptasensor is 2 nM, which is lower than the reported ThT/ATP aptamer based methods. Additionally, our aptasensor has a wider dynamic linear range (0.1 µM - 120 µM) and higher selectivity. The proposed aptasensor has been successfully applied to detect ATP in 15% human serum. More importantly, the current study not only provides a novel method for ATP assay but also presents a way to construct a label-free split aptamer based fluorescent sensor for other species where aptamer can be generated.

In Situ Generation of Fluorescent Copper Nanoclusters Embedded in Monolithic Eggshell Membrane: Properties and Applications.

Luminescent metal nanoclusters have attracted considerable research attention in recent years due to their unique properties and extensive usage in many fields. Three different synthetic routes were developed to in situ generate orange and red emitting copper nanoclusters embedded in monolithic eggshell membrane (Cu NCs@ESM) using different reducing reagents including N2H4·H2O, NH2OH·HCl and Vitamin C at room temperature for the first time. The routes are extremely facile, low-cost and versatile. The obtained Cu NCs@ESM nanocomposites exhibit excellent photostability and chemical stability, laying the foundation for various practical applications. Fluorescent surface patterning was demonstrated based on the proposed strategy easily. Significantly, the Cu NCs@ESM shows selective fluorescence quenching response to Hg2+ ions and good catalytic activity for methylene blue (MB) reduction degradation making it ideal as portable sensing strip and recyclable catalyst. The work provides a general strategy for the fabrication of other various monolithic nanomaterials with potential applications.

Visual detection of sub-femtomole DNA by a gold nanoparticle seeded homogeneous reduction assay: toward a generalized sensitivity-enhancing strategy.

Gold nanoparticles (AuNPs) have been employed to design colorimetric visual sensing assays toward the detections of various targets including DNA, based on the aggregation induced color transitions of AuNPs. However, the relatively high detection limit (LOD 10nM) in the case of DNA detection has become a stumbling block on the road of the further development and applications of these assays. This research aims at overcoming this limit by virtue of a seeded gold reduction strategy. Typically, low concentrations of 13 nm AuNPs modified with suitable DNA probes are allowed to hybridize with a DNA target to form aggregates, which are then transferred into a gold-enhancing cocktail for further depositions of more gold on the initial AuNP seeds. The color of the assay thus sensitively reflects the initial aggregation status of the 13 nm AuNPs, which can be related to the concentrations of the DNA target. This assay has a sensitivity that is at least 25-50 times improved. Under still not fully optimized conditions, 0.4 fmol DNA in a 2 microL sample can be confidently detected with the ability of distinguishing a single base mutation. It does not require isolations of the 13 nm AuNP aggregates during analyses and shares the advantages of a homogeneous assay, including simplicity, adaptability, convenience, and being free of interferences due to non-specific surface adsorptions.

Colorimetric Hg2+ detection with a label-free and fully DNA-structured sensor assembly incorporating G-quadruplex halves.

A sensitive and fully DNA-structured ion sensor was built by integrating polyT sequences for highly selective Hg2+ recognitions and two flanking G-quadruplex halves for allosteric signal transductions. The construction of this sensor was very easy that allowed a cost-effective detection of Hg2+ with a limit of detection of 4.5 nM, which was lower than the 10 nM toxic level for drinkable water as regulated by the US's EPA. The strategy employed for the construction of this sensor may be further extended to other sensors through a rational structural fusion between re-engineered aptameric and enzymic DNA sequences.

Rational design of an optical adenosine sensor by conjugating a DNA aptamer with split DNAzyme halves.

Split halves of a hemin-binding DNAzyme have been assembled with an anti-adenosine aptamer to build a homogeneous allosteric sensor for adenosine with high selectivity and sensitivity.