Fluorescence Response Research Articles

Rapid Fluorescence Assay for Polyphosphate in Yeast Extracts Using JC-D7.

Polyphosphate (polyP) is an intriguing molecule that is found in almost any organism, covering a multitude of cellular functions. In industry, polyP is used due to its unique physiochemical properties, including pH buffering, water binding, and bacteriostatic activities. Despite the importance of polyP, its analytics is still challenging, with the gold standard being 31P NMR. Here, we present a simple staining method using the fluorescent dye JC-D7 for the semi-quantitative polyP evaluation in yeast extracts. Notably, fluorescence response was affected by polyP concentration and polymer chain length in the 0.5-500 µg/mL polyP concentration range. Hence, for polyP samples of unknown chain compositions, JC-D7 cannot be used for absolute quantification. Fluorescence of JC-D7 was unaffected by inorganic phosphate up to 50 mM. Trace elements (FeSO4 > CuSO4 > CoCl2 > ZnSO4) and toxic mineral salts (PbNO3 and HgCl2) diminished polyP-induced JC-D7 fluorescence, affecting its applicability to samples containing polyP-metal complexes. The fluorescence was only marginally affected by other parameters, such as pH and temperature. After validation, this simple assay was used to elucidate the degree of polyP production by yeast strains carrying gene deletions in (poly)phosphate homeostasis. The results suggest that staining with JC-D7 provides a robust and sensitive method for detecting polyP in yeast extracts and likely in extracts of other microbes. The simplicity of the assay enables high-throughput screening of microbes to fully elucidate and potentially enhance biotechnological polyP production, ultimately contributing to a sustainable phosphorus utilization.

Fluorescence and Circular Dichroism Dual-Mode Probe for Chiral Recognition of Tyrosine and Its Applications in Bioimaging.

Chiral amino acids (AAs) are essential in metabolism and understanding physiological processes, and they could be used as biomarkers for the diagnosis of different diseases. In this study, chiral Cdots@Van were prepared by postmodifying an achiral Cdots core with vancomycin for recognizing and determining the enantiomeric excess (ee) of tyrosine (Tyr) enantiomers. The fluorescence response of Cdots@Van is based on an "on-off" strategy, with different quenching percentages for d- and l-tyrosine. Interestingly, the circular dichroism (CD) spectrum of Cdots@Van responded to only one form of Tyr enantiomer, specifically d-Tyr, and remained nearly unchanged upon the addition of l-Tyr. Quantum mechanical (QM) calculations were in excellent agreement with the experimental results, confirming the stronger binding affinity of Cdots@Van for d-Tyr compared to l-Tyr. We further investigated the chiral recognition ability of the interconnected vancomycin particles, which was synthesized using the EDC/NHS coupling reaction between vancomycin molecules without a Cdots core. Surprisingly, unlike free vancomycin molecules, interconnected vancomycin displayed an enantiomeric recognition ability by CD spectroscopy, similar to what was observed for Cdots@Van. Crucially, this chiral probe has been successfully utilized for cell imaging applications.

Sensitive and selective fluorescence detection of acetamiprid using oxidized single-walled carbon nanohorns and cryonase-assisted signal amplification

To address the low accuracy of pesticide residue detection in traditional Chinese medicine, we developed a biosensor for acetamiprid detection. First, an oxidized single-walled carbon nanohorn capable of adsorbing and quenching fluorescence was prepared. Second, a novel fluorescence sensing system was constructed based on this nanomaterial. The detection performance of the sensor system was evaluated, which revealed that fluorescence quenching by the biosensor was primarily due to the adsorption of the aptamer by the nanomaterials. Optimal conditions were found to be a nanomaterial mass concentration of 800 ng/mL and an incubation time of 70 min, which resulted in the best fluorescence response. Under these optimal conditions, the system was used to detect acetamiprid residues in traditional Chinese medicine. The fluorescence intensity showed a strong linear relationship with acetamiprid concentration, with a correlation coefficient of 0.9985 and a detection limit of 6.33 ng/mL. In the sample test, the biosensor demonstrated high accuracy, with good average recovery rates and low relative standard deviation. The system showed excellent selectivity and detection performance, with no interference from other pesticides.

Constructing high efficient carrier channels in double ternary compounds MoSSe/CdSSe heterojunction for photoelectrochemistry and electrocatalytic hydrogen production

Here is a demonstration of constructing MoSSe/CdSSe heterojunction catalyst from bimetallic selenium sulfides. CdSSe has a high conduction band position of −0.65 eV, resulting in a strong reduction potential of photogenerated electrons. However, the high resistance of CdSSe affects the transport of charge carriers. In addition, experiments have confirmed that the presence of S and Se in MoSSe and CdSSe enhances their chemical activity and facilitates bonding. A better connection can be formed between MoSSe and CdSSe, resulting in high carrier transport efficiency. In the X-ray photoelectron spectroscopy results, the Mo and Se signals showed a large shift nearly 1 eV which confirms the strong binding between MoSSe and CdSSe. In the photoelectrochemical tests, the MoSSe/CdSSe heterojunction demonstrated a photocurrent density of 3.097 mA/cm2 at 0 V (RHE) which is 2.7 and 2.9 times that of MoSSe (1.227 mA/cm2) and CdSSe (1.127 mA/cm2) respectively with a low resistance of 161 Ω. Moreover, the MoSSe/CdSSe also has good electrocatalytic hydrogen evolution performance with an overvoltage of 180 mV and Tafel slope of 69 mV/dec. Fluorescence and photocurrent response tests confirmed its higher carrier separation efficiency. This work demonstrates the application of ternary metal selenium sulfide heterojunctions in catalytic energy materials.

Hierarchical Bi2Fe4O9/BiOI S-scheme heterojunctions with exceptional hydraulic shear induced photo-piezoelectric catalytic activity

Hierarchical Bi2Fe4O9/BiOI S-scheme nanoflower heterostructures are prepared by hydrothermal method, which exhibit exceptional photo-piezoelectric catalytic performance. The tight binding between the sheets ensures the efficient electron transport, and provides a large interface area and adequate reaction sites for photo-piezoelectric catalytic reactions. At the same time, because the water flow in the water body produces hydraulic shear force on the material, the material produces piezoelectric effect. Bi2Fe4O9/BiOI exhibit a remarkable degradation efficiency of 99.4% for tetracycline and a hydrogen production rate of 4089.36 µmol h−1 g−1. The observed behavior can be explained by the combined influence of the formation of S-scheme structure and the process of photo-piezoelectric catalysis, confirmed by in-situ XPS, transient/steady-state fluorescence and piezoelectric response force test. The excellent stability of the material suggests its possible use in the sectors of energy and environment. This work introduces novel concepts for the future advancement of photo-piezoelectric synergistic catalysis.

Chitosan based fluorescent probe with AIE property for detection of Fe3+ and bacteria

Fluorescent probe with aggregation-induced emission (AIE) property has been widely used because of the advantages of high sensitivity, good selectivity and non-destructive testing. The development of fluorescent probe with good biocompatibility, photostability and biodegradability is of great significance in biomedicine and environmental detection. Herein, a novel type of fluorophore CS-TPE for detection of Fe3+ and bacteria was prepared by the Schiff base reaction of chitosan (CS) and 4-(1,2,2-triphenylethenyl) benzaldehyde (TPE-CHO). The fluorescence response mechanism of CS-TPE system was investigated by various characterization techniques. CS-TPE had an obvious AIE behavior with strong blue-green emissions at 473 nm and reaches the highest photoluminescence (PL) emission in 90 % H2O/ethanol mixtures. CS-TPE fluorescent probe exhibited sensitive quenching response to Fe3+, which can be used as a biosensor for detecting the concentration of Fe3+ with short response time (5 min), low detection limit (0.998 μM) and wide detection range (10–300 μM). Meanwhile, CS-TPE exhibited good antibacterial performance for S. aureus and E. coli. It is expected to realize the real-time fluorescence monitoring of metal ion detection and antibacterial process.

A novel fluorescence platform for portable and visual monitoring of meat freshness

Biogenic amines (BAs) are crucial markers of meat spoilage. Developing practical and effective BAs detection methods is essential for monitoring meat freshness and ensuring daily consumption safety. This study prepared several naphthalene-based fluorescent compounds to visually monitor meat freshness in real-time. These probes show a colorimetric fluorescence response to putrescine and cadaverine (typical spoilage indicators) through nucleophilic addition/elimination reaction. The detectability of these probes can be optimized by altering the electronegativity and substitution position of the recognition group. Among these compounds, 2-((6-(4-(diphenylamino)phenyl)naphthalen-2-yl)methylene)malono nitrile (TNMA) demonstrated exceptional sensing performance toward putrescine and cadaverine, including high-contrast fluorescence color transition (red to blue), rapid response times (∼30 s), high selectivity and sensitivity (detection limit for putrescine: 2.69 ppm, cadaverine: 6.11 ppm). Furthermore, the B/R values of TNMA test strips output by RGB analysis presented a linear correlation with total volatile basic nitrogen (TVBN, an international standard for evaluating food spoilage) values in pork. Based on this correlation, we utilized smartphone applications to construct an intelligent evaluation system, enabling visual monitoring of pork, chicken, and shrimp freshness under various storage conditions. The TNMA-based system offers a reliable platform for real-time, portable and visual monitoring of meat freshness for consumers and suppliers in the food industry.

A silver auto-catalyzed plasmonic enzyme-linked immunosorbent assay for colorimetric and fluorescent detection of neutrophil gelatinase associated lipocalin (NGAL)

The conventional ELISA lacked sufficient sensitivity and accuracy for analyzing neutrophil gelatinase associated lipocalin (NGAL), a potential biomarker of ulcerative colitis. Here, we developed a dual-readout plasmonic ELISA with cascade amplification of alkaline phosphatase hydrolysis and O-phenylenediamine oxidation. In this novel plasmonic ELISA, alkaline phosphatase-conjugated antibodies capture NGAL from samples, alkaline phosphatase hydrolyzes 2-phospho-L-ascorbic acid into ascorbic acid, and then ascorbic acid blocks silver-catalyzed O-phenylenediamine oxidation by turning silver ions into silver microwires. Therefore, NGAL concentration is proportional to the decrease in chromogenic and fluorescent response. This novel plasmonic ELISA exhibited a linear range of 0.5–80 ng/mL, and the LOD was as low as 0.5 ng/mL. The sensitivity was superior to that of conventional ELISA and previous studies. The plasmonic ELISA exhibited favorable selectivity because of the specific antibody-antigen interaction and the robust cascade amplification. Besides, the dual-readout feature endowed the plasmonic ELISA with advantages in terms of convenience and compatibility. In summary, our study provided a novel and sensitive plasmonic ELISA for the diagnosis of ulcerative colitis.

Selective cyanide ion detection using hydrazine carbothioamide probe: A fluorescence turn-on approach

This study reports the synthesis of various hydrazine carbothioamide derivatives through a Schiff base reaction, confirmed by FT-IR, NMR (1H and 13C), and HRMS spectroscopic methods. Among the synthesized derivatives, probe L ((4-fluorobenzylidene)-2-(1-phenylethylidene)hydrazine-1-carbothioamide) was identified as a highly selective fluorometric chemosensor based on its absorbance and emission spectra. Probe L showed a CN− ion-specific turn-on fluorescence response, with very little interference from other anions. Job's method established a 1:1 binding stoichiometry among probe L and the cyanide ion. The detection limits (LOD) and quantification (LOQ) were observed to be 0.639 µM and 2.116 µM, respectively. With the use of FT-IR spectroscopy, 1H NMR titration, 13C NMR and HRMS, the binding mechanism was clarified. Moreover, probe L was employed effectively for detecting CN− ions in water and food samples, highlighting its potential as a practical sensor for cyanide detection.

A highly specific red-emitting fluorescent probe for monitoring of Cu2+ in living cells and zebrafish.

A novel seminaphthorhodafluor-based fluorescent probe was designed and prepared. It exhibited an outstanding off-on fluorescence response (about 45-fold enhancement) to Cu2+ because the structure changed from a spirocyclic form to a ring-open state. It showed satisfactory detection performances for Cu2+ with fast response speed, high selectivity and anti-interference, and a low detection limit (22.9nM) due to its long emission wavelength and off-on fluorescence response mode. This probe was also satisfactorily applied for the monitoring of Cu2+ in living cells and zebrafish.

Mechanism study of Dual-Emission ratiometric fluorescent pH-Sensitive carbon quantum dots and its application on mornitoring enzymatic catalysis

Carbon dots (CQD) have received significant attention as a novel ratiometric fluorescent pH nanoprobe, owing to their favorable optical properties and excellent biocompatibility. Despite their appealing features, the precise mechanism behind the pH-sensitive photoluminescence of CQDs remains to be fully understood. This study endeavors to unravel the mechanism underlying the pH-responsive ratiometric fluorescence in dual-emission CQDs, synthesized through a one-step hydrothermal method using o-phenylenediamine and oxalic acid as precursors. The resultant CQDs exhibit inherent dual-emission at wavelengths of 383 nm and 566 nm, with the ratiometric fluorescence response tailored by the ratio of precursors, providing a robust tool for pH sensing across a range of 2 to 6. Detailed characterizations, including chemical, morphological, and optical analyses, alongside theoretical insights from time-dependent density functional theory (TD-DFT), elucidate the mechanism underlying the pH-dependent luminescence, attributed to the electron cloud transmission between amide and adjacent carboxyl groups on the CQD surface. The superior performance of these CQDs in real-time pH monitoring is demonstrated through their application in glucose oxidase-catalyzed reactions, showcasing their potential as efficient, reliable nanoprobes for biomedical research and diagnostic applications.

Fluorescent probe for imaging N2H4 in plants, food, and living cells and for quantitative detection of N2H4 in soil and water using a smartphone

Hydrazine is volatile and highly toxic, causing severe harm to water, soil, air, and organisms. Therefore, real-time detection and long-term monitoring of hydrazine are crucial for environmental protection and human health. Herein, an “OFF−ON” fluorescent probe 5-((10-ethyl-2-methoxy-10 H-phenothiazin-3-yl)methylene)−2,2-dimethyl-1,3-dioxane-4,6-dione (MPD) for hydrazine detection through a nucleophilic addition reaction was developed. MPD could exclusively identify hydrazine through colorimetric and fluorescent dual-channel responses within 30 s, which also demonstrated high sensitivity (detection limit, 12 nM) and a wide pH range (6 −12). The sensing mechanism of MPD was confirmed using theoretical calculations, where fluorescence was emitted following the recognition of hydrazine because of the disappearance of the photoinduced electron transfer (PET) process. Using a smartphone, MPD enabled the quantitative detection of hydrazine in real water samples and sandy soil. Notably, in the process of detecting hydrazine in actual water samples, the establishment of analytical methods and the completion of rapid quantitative detection only required a smartphone and built-in apps. Additionally, we showed that MPD could recognize hydrazine in various environmental samples, including plants, food, hydrazine vapors, and cells. We believe that the fluorescent probe MPD developed in this study and the established smartphone visualization platform will provide a convenient and effective tool for detecting hydrazine in environmental monitoring, food safety assessment, biological system safety, and other fields.

Unlocking lysosomal acidity to activate membranolytic module for accurately cancer theranostics

Chemotherapy drug efflux, toxic side effects, and low efficacy against drug-resistant cells have plagued safe and efficient cancer theranostics. However, the materials or methods that resolve these defects all-in-one are scarce. Here, a new cancer theranostics strategy is proposed by utilizing changes in lysosomal acidity in cancer cells to activate the membranolytic model to overcome these obstacles together. Therefore, a simple fluorescent anthracene derivative Lyso-Mito is developed, which has a perfect pKa (4.62) value that falls between the pH of lysosomes in cancer and normal cells. Lyso-Mito itself can precisely target and convert the pH perturbation of lysosomes in cancer cells to fluorescent response and membranolytic module activity to accomplish the low drug efflux, weak toxic side effects, and low drug-resistant cancer diagnosis and treatment without linking other functional units or any additional assistance. Hereby, a new cancer theranostics strategy of integrating organelle microenvironment and the membranolytic model is realized.

A ratiometric fluorescent probe for the detection of γ-glutamyl transpeptidase activity and its application in garlic

A novel ratiometric fluorescent probe for the detection of γ-glutamyl transpeptidase (GGT) activity was developed. The recognition of GGT by this probe is achieved by its enzymatic hydrolysis of peptide bonds in the probe. Under optimized conditions, the probe exhibited obvious ratiometric fluorescence responsiveness after interaction with GGT, avoiding potential background interference. The linear concentration range was 0.0350–2 U / mL, with good selectivity and stability. Under a 365 nm UV lamp, as the concentration of GGT increased, the probe aqueous solution gradually changed from dark blue to light blue. In addition, the B/G value of the probe's solution and the GGT activity were linearly correlated. More significantly, the probe was able to effectively identify the GGT activity of garlic.

Unlocking multi-mode sensing potential: Phosphorus-doped graphitic carbon nitride quantum dots for Ag+, ciprofloxacin, and riboflavin analysis in environment and food matrices

The simultaneous detection of multiple analytes through a single fluorescence sensor is highly attractive. In this study, phosphorus-doped graphitic carbon nitride quantum dots (P-CNQDs) were developed, achieving multi-mode sensing through three distinct response mechanisms. The preparation involved using melamine as the carbon and nitrogen source and ammonium dihydrogen phosphate as the phosphorus source. Uniform and narrowly distributed P-CNQDs were successfully synthesized through chemical oxidation and hydrothermal methods, with an average size of 2.4 nm. These unique P-CNQDs exhibited fluorescence quenching through photo-induced electron transfer (PET) in response to Ag+. Additionally, the formation of hydrogen bonds and coordination interactions between P-CNQDs-Ag+ and ciprofloxacin (CIP) led to a pronounced fluorescence response to CIP by the chelation enhanced fluorescence (CHEF) mechanism. Furthermore, leveraging the principle of fluorescence resonance energy transfer (FRET), P-CNQDs-CIP served as a ratio fluorescence sensor for riboflavin (RF), enabling ultra-sensitive detection of RF. The combination of PET, CHEF, and FRET response mechanisms successfully facilitated multi-mode sensing for Ag+, CIP, and RF. The detection ranges were 0.05–100 μM, 0.002–2 μM, and 0.05–60 μM, with corresponding lowest detection limits of 17.1 nM, 1.1 nM, and 29.2 nM, respectively. This versatile sensor has been effectively applied to real samples, including the detection of river water and vitamin B2 tablets.

Construction of HaloTag-based macromolecular probe for multiple logic gates and photoactivatable bioimaging

Protein bioconjugation has emerged as one of the most valuable tools for the development of protein-based biochemical assays. Herein, we report a fluorescent macromolecular probe RF12_POI, in which the coumarin derivative RF12 is specifically conjugated onto the HaloTag fused protein of interest (POI) to achieve a dual stimuli-mediated fluorescence response. RF12 is first obtained by installing a photo-cleavable 1-ethyl-2-nitrobenzyl group onto the C7 hydroxy moiety of coumarin fluorophore with a HaloTag ligand attaching to the acid-labile 1,3-dioxane moiety. Upon stimulation, RF12_Halo exhibits a sequential fluorescence response to photon/H+ on both liquid and solid interfaces. Through the conjugation of RF12 onto the GFP_Halo protein, RF12_GFP_Halo presents a fluorescence resonance energy transfer (FRET) from photo-cleaved RF12 to GFP in the protein complex. Furthermore, by utilizing the stimuli-responsive fluorescence characteristics of coumarin derivatives RF12 (photon/H+) and RF16 (H2O2/H+), we construct RF12/RF16_POI based protein films and achieve multiple applications of logic circuits, including AND, OR, XOR, INHIBIT, Half-adder or Half-subtractor. In these circuits, the output value of I/I0 is dependent on the input sequence of photon, H2O2, and H+. Additionally, we evaluate the fluorescence labeling ability of RF12 to intracellular IRE1_Halo protein and demonstrate that RF12 containing the HaloTag ligand could be precisely retained in cells to track IRE1_Halo protein. Hence, we provide a unique structural design strategy to construct fluorescence dual-responsive macromolecules for information encryption and cellular protein visualization.

A dual-channel fluorescent probe targeting lysosomes for differential detection of Cys/Hcy and GSH: Applications in food, pharmaceutical analysis and bioimaging

Thiols function as antioxidants in food, prolonging shelf life and enhancing flavor. Moreover, thiols are vital biomolecules involved in enzyme activity, cellular signal transduction, and protein folding among critical biological processes. In this paper, the fluorescent probe PYL-NBD was designed and synthesized, which utilized the fluorescent molecule pyrazoline, the lysosome-targeted morpholine moiety, and the sensing moiety NBD. Probe PYL-NBD was tailored for the recognition of biothiols through single-wavelength excitation, yielding distinct fluorescence emission signals: blue for Cys, Hcy, and GSH; green for Cys, Hcy. Probe PYL-NBD exhibited rapid reaction kinetics (<10 min), distinct fluorescence response signals, and low detection limits (15.7 nM for Cys, 14.4 nM for Hcy, and 12.6 nM for GSH). Probe PYL-NBD enabled quantitative determination of Cys content in food samples and L-cysteine capsules. Furthermore, probe PYL-NBD had been successfully applied for confocal imaging with dual-channel detection of biothiols in various biological specimens, including HeLa cells, zebrafish, tumor sections, and Arabidopsis thaliana.

Modulating Fluorescent Responses in Organic-Doped Polyethylenimine Composites: Impact on Pyrophosphate Detection and Biogenic Phosphate Analysis

Modulating Fluorescent Responses in Organic-Doped Polyethylenimine Composites: Impact on Pyrophosphate Detection and Biogenic Phosphate Analysis

An Integrated Polysaccharide Hydrogel with Versatile Fluorescence Responses through Noncovalent Reformation of Gel Aggregation and for Bioimaging.

Functionalized hydrogels, with their unique and adaptable structures, have attracted significant attention in materials and biomaterials research. Fluorescent hydrogels are particularly noteworthy for their sensing capabilities and ability to mimic cellular matrices, facilitating cell infiltration and tracking of drug delivery. Structural elucidation of hydrogels is crucial for understanding their responses to stimuli such as the pH, temperature, and solvents. This study developed a fluorescent hydrogel by functionalizing chitosan with p-cresol-based quinazolinone aldehyde. Confocal microscopy revealed the hydrogel's intriguing fluorogenic properties. The hydrogel exhibited enhanced fluorescence and a tunable network morphology, influenced by the THF-water ratio. The study investigated the control of gel network reformation in different media and analyzed the fluorescence responses and structural changes of the sugar backbone and fluorophore. Proper selection of mixed solvents is essential for optimizing the hydrogel as a fluorescence probe for bioimaging. This hydrogel demonstrated greater swelling properties, making it highly suitable for drug delivery applications.

Gold Nanocluster-Based Self-Assembly Fluorescence Microbeads for Sensor Array Discrimination of Multicomponent Metal Ions.

Benefiting from easy visualization and simultaneous detection of multiple targets, fluorescence microbeads are commonly used as fluorescence-sensing elements to detect pollutants in the environment. However, the application of fluorescence microbead-based sensor arrays is still limited because fluorescence dyes always suffer from self-quenching, photobleaching, and spectral overlap. Herein, three kinds of gold nanoclusters (Au NCs) were assembled with polystyrene microspheres (PS NPs) by electrostatic interaction to prepare fluorescence microbeads (PS-Au NCs), developing a sensor array for the simultaneous analysis of multiple metal ions. In this work, different PS-Au NCs showed an enhancing or quenching fluorescence response to various metal ions, owing to distinct binding capacities. Combined with the recognition algorithm from linear discriminant analysis (LDA) and hierarchical cluster analysis (HCA), this sensor assay could realize single-component and multicomponent qualitative detection for 8 kinds of heavy metal ions (HMIs) including Cu2+, Co2+, Pb2+, Hg2+, and Ce3+. Particularly, the large surface area of PS NPs could provide a direct reaction microenvironment to improve the efficiency of the detection process. Meanwhile, the fluorescence property of Au NCs could also be enhanced by a partially effective aggregation-induced emission (AIE) effect to give better fluorescence signal output. Under optimal conditions, 8 kinds of heavy metals and their multicomponent mixtures could be identified at concentrations as low as 0.62 μM. Meanwhile, the analytical performance of this sensor assay in water samples was also verified, meeting the requirement of actual analysis. This study provides a great potential and practical example of single-batch, multicomponent identification for HMIs.