Acid Fluorescent Probes Research Articles

Highly-Selective fluorescent Fe3O4@PPy aptasensor

Label-free nucleic acid fluorescent probes are gaining popularity due to their low cost and ease of application. However, the primary challenges associated with label-free fluorescent probes stem from their tendency to interact with other biomolecules, such as RNA, proteins, and enzymes, which results in low specificity. In this work, we have developed a simple detection platform that utilizes Fe3O4@PPy in combination with a label-free nucleic acid probe, 1,1,2,2-tetrakis[4-(2-bromo-ethoxy)phenyl]ethene (TTAPE) or Malachite Green (MG), for highly selective detection of metal ions, acetamiprid, and thrombin. Fe3O4@PPy not only adsorbs aptamers through electrostatic interactions, π–π bonding, and hydrogen bonding, but also quenches the fluorescence of the TTAPE/MG. Upon the addition of target compounds, the aptasensor separates from Fe3O4@PPy through magnetic separation. Moreover, by changing different aptamers, the aptasensor was applied to detect metal ions, acetamiprid, and thrombin, with the turned-on photoluminescence (PL) emission intensity recorded and showing linearity to the concentrations of targets. The robustness of method was demonstrated by applying it to real samples, which included vegetables (for detecting acetamiprid with LODs of 0.02 and 0.04 ng/L), serum samples (for detecting thrombin with LODs of 5.5 and 4.3 nM), and water samples (for detecting Pb2+ with an LOD of 0.17 nM). Therefore, due to its impressive selectivity and sensitivity, the Fe3O4@PPy aptasensor could be utilized as a universal detection platform for various clinical and environmental applications.

A single 8-hydroxyquinoline-appended bile acid fluorescent probe obtained by click chemistry for solvent-dependent and distinguishable sensing of zinc(II) and cadmium(II).

Construction of fluorescent probes for zinc ion (Zn2+ ) and cadmium ion (Cd2+ ) is significant for the safety of humans. However, the discriminating recognition of Zn2+ and Cd2+ by a single probe remains challenging owing to their similar properties. Herein, a novel deoxycholic acid derivative containing 8-hydroxyquinoline fluorophore has been facilely synthesized through click chemistry to form a clamp-like probe. Using its perfect bonding cavity from 1,2,3-triazole and quinoline, this molecule showed favorable solvent-dependent fluorescent responses and distinguished Zn2+ and Cd2+ in different solvents. In ethanol aqueous solution, it displayed good selectivity and ratiometric fluorescence to Zn2+ with 30 nm spectroscopic red-shifts. In acetonitrile aqueous solution, it exhibited good selectivity and ratiometric fluorescence to Cd2+ with 18 nm spectroscopic red-shifts. Moreover, the unique microstructural features of the probe in assembly were used to reflect its recognition processes. Due to its merits of low detection limit and instant response time, the probe was utilized for sensing Zn2+ and Cd2+ in water, beer and urine with high accuracy. Meanwhile, this probe served as a handy tool and was employed to obtain inexpensive test strips for the prompt and semiqualitative analysis of Zn2+ and Cd2+ with the naked eye.

Rapid detection of prostate cancer biomarkers in living cells and urine samples using diboronic acid fluorescent probe

Rapid and sensitive screening methods are critical for early-stage cancer diagnosis. Sialyl Lewis X (sLeX), a terminal O-glycan structure highly expressed on the surface of cancer cells, can be employed as a marker for cancer detection. In this study, a fluorescence analysis method based on a diboronic acid fluorescent (DBF) probe was established to detect the content of sLeX in solution under physiological conditions within 10 min and sLeX-carrying secreted proteins in prostate cancer cells within 15 min, with a limit of detection of 4.58 × 10−10 mol/L. Fluorescence microscopy imaging confirmed that proteins carrying sLeX were secreted from the cell into the extracellular space. Finally, urine samples from clinical patients (n = 15) and healthy volunteers (n = 5) were examined and analyzed using this method, and significant differences in sLeX levels between the two groups were found. In conclusion, we successfully established a method for detecting sLeX levels in urine samples of patients with prostate cancer using a DBF probe, which can be used to improve early non-invasive screening, disease development monitoring and prognostic detection of prostate cancer.

Acidic tumor microenvironment-activatable fluorescent diagnostic probe for the rapid identification and resection of human tumors via spraying

A fluorescent diagnostic probe for real-time intraoperative image-guided tumor resection can significantly improve the efficiency and quality of oncological therapy, but their development is challenging. Herein, a novel fluorescent diagnostic probe called HLTC based on β-carboline was designed and synthesized. HLTC was found to show a ∼10-fold enhancement of fluorescence quantum field with pH from 7.4 to 4.0, indicating its imaging potential in acid environment which is a typical hallmark of the tumor microenvironment (TME). Following fluorescence microscopy imaging showed HLTC could emit specific signals in cancer cells and sections, by both one-photon excitation and two-photon excitation. Importantly, HLTC enabled the precise and rapid delineation of both transplanted tumor and clinical tumor tissues within several minutes of simple topical spray. The tumor-to-background ratio (TBR) was up to 10.2 ± 1.0 at clinical liver cancer tissues and 9.9 ± 0.3 at clinical colon cancer tissues, allowing precise tumor margin identification and the effective guidance of surgical tumor resection. Furthermore, CCK8 assay, pharmacokinetic evaluation, blood analysis and H&E staining were performed, which verified high biocompatibility and biosafety of HLTC at working concentration. These results reveal the exciting potential of this small-molecule fluorescent diagnostic probe for real-time fluorescence-based navigation during surgical tumor resection.

Molecular Dynamics of Glycolipids in Liposomes.

Glycosphingolipids (GSLs) in the mammalian plasma membrane are essential for various biological events as they form glycolipid-rich membrane domains, such as lipid rafts. GSLs consist of a certain oligosaccharide head group and a ceramide tail with various lengths of acyl chains. The structure of the head group as well as the carbon number and degree of the unsaturation of the acyl chain are known to regulate the membrane distributions and interleaflet couplings of GSLs by altering physicochemical properties, such as dynamics, interactions, and cluster sizes. This chapter provides the detailed use of time-resolved fluorescence measurement for investigating the membrane properties of lactosylceramide (LacCer)-enriched domains in bilayer membranes. LacCer belongs to the neutral GSLs and is believedin forming a highly ordered phase in model membranes and biological membranes, while the details of the domain remain unclear. Here, we suggest using trans-parinaric acid (tPA) and tPA-LacCer fluorescent probes to reveal the dynamics and size of the GSL domains since they prefer to bedistributed in the GSL-rich ordered phase. The fluorescence lifetime in the nanosecond timescale reveals the difference in the surrounding membrane environments, which relates to hydrocarbon chain ordering, membrane hydration, and submicrometer domain size. The fluorescence lifetime of these probes can thus provide important information on submicron- to nano-scale small GSL domains not only in model membranes but also in biological membranes.

A triphenylamine-based fluorescent probe with phenylboronic acid for highly selective detection of Hg2+ and CH3Hg+ in groundwater.

Mercury is a highly toxic heavy metal and it poses a serious threat to the natural environment and human health. Thus, selective detection of trace mercury (e.g. inorganic mercury and methylmercury) in the environment is critical yet challenging. Herein, we describe the rational design and facile synthesis of a new triphenylamine-based phenylboronic acid fluorescent probe (TPA-PBA) for selective detection of Hg2+ and CH3Hg+. Due to the inherent specificity of the displacement reaction between phenylboronic acid and mercury, this probe exhibits exceptionally high selectivity towards Hg2+/CH3Hg+ against other tested ions with ppb-level sensitivity. More importantly, the probe TPA-PBA is effective and selective in detecting Hg2+/CH3Hg+ in tap water and real-world groundwater, indicating its potential practical applications in in situ and online mercury detection in real-world scenarios. With TPA-PBA based test strips Hg2+ can be distinguished from CH3Hg+ by the naked eye. This study could accelerate the development of low-cost, highly efficient and selective fluorescent probes for rapid trace mercury detection.

Directed evolution engineering to improve activity of glucose dehydrogenase by increasing pocket hydrophobicity.

Glucose dehydrogenase (GDH) is a NAD(P)+ dependent oxidoreductase, which is useful in glucose determination kits, glucose biosensors, cofactor regeneration, and biofuel cells. However, the low efficiency of the catalysis hinders the use of GDH in industrial applications. In this study, an analysis of interactions between eight GDH mutants and NADP+ is powered by AlphaFold2 and Discovery Studio 3.0. The docking results showed that more hydrogen bonds formed between mutants, such as P45A and NADP+, which indicated that these mutants had the potential for high catalytic efficiency. Subsequently, we verified all the mutants by site-directed mutagenesis. It was notable that the enzyme activity of mutant P45A was 1829 U/mg, an improvement of 28-fold compared to wild-type GDH. We predicted the hydrophobicity of the protein-ligand complexes, which was confirmed by an 8-anilino-1-naphthalenesulphonic acid fluorescent probe. The following order of increasing hydrophobicity index was deduced: GDH < N46E < F155Y < P45A, which suggested that the enzyme activity of GDH is positively related to its pocket hydrophobicity. Furthermore, P45A still showed better catalytic ability in organic solvents, reaching 692 U/mg in 10% isopropanol, which was 19-fold that of the wild-type GDH. However, its substrate affinity was affected by organic solvents. This study provides a good theoretical foundation for further improving the catalytic efficiency of GDH.

Oxidative leaching of V-Cr–bearing reducing slag via a Cr(III) induced Fenton-like reaction in concentrated alkaline solutions

V-Cr–bearing reducing slag (VCRS) is considered a hazardous waste that can create ecosystem disasters if handled improperly. It consists of a considerable amount of heavy metals, such as vanadium (V) and chromium (Cr). In this study, we propose a novel process featuring a VCRS self-induced Cr(III)-Fenton-like reaction to efficiently recover V and Cr from hazardous VCRS. The generation of hydroxyl radicals (·OH) and determination of their effect on V and Cr oxidation were examined via electron spin resonance detection, free radical quenching, and terephthalic acid fluorescence probe methods. The V and Cr oxidative leaching processes were directly controlled by the amount of added H2O2 and generated·OH from the Cr(III)-Fenton-like reaction, which in turn was dependent on the amount of dissolved Cr(OH)4-. In a single oxidative leaching process, the leaching efficiencies of V and Cr reached 97.5 ± 0.6 % and 85.2 ± 0.8 %, respectively, and reached 99.4 ± 0.5 % and 94.6 ± 0.9 %, respectively, from circular leaching owing to a continuous supply of dissolved Cr(OH)4- from fresh VCRS. This study identifies a novel approach to discovering deep oxidation of the VCRS while minimizing environmental contamination via a waste control strategy and can be considered an attractive alternative approach for the green treatment of VCRS.

Amplification of the Fluorescence Signal with Clustered Regularly Interspaced Short Palindromic Repeats-Cas12a Based on Au Nanoparticle-DNAzyme Probe and On-Site Detection of Pb2+ Via the Photonic Crystal Chip.

Although great headway has been made in DNAzyme-based detection of Pb2+, its adaptability, sensitivity, and accessibility in complex media still need to be improved. For this, we introduce new ways to surmount these hurdles. First, a spherical nucleic acid (SNA) fluorescence probe (Au nanoparticles-DNAzyme probe) is utilized to specifically identify Pb2+ and its suitability for precise detection of Pb2+ in complex samples due to its excellent nuclease resistance. Second, the sensitivity of Pb2+ detection is greatly enhanced via the use of a clustered regularly interspaced short palindromic repeats-Cas12a with target recognition accuracy to amplify the fluorescent signal upon the trans cleavage of the SNA (signal probe), and the limit of detection reaches as low as 86 fM. Third, we boost the fluorescence on photonic crystal chips with a bionic periodic arrangement by employing a straightforward detection device (smartphone and portable UV lamp) to achieve on-site detection of Pb2+ with the limit of detection as low as 24 pM. Based on the abovementioned efforts, the modified Pb2+ fluorescence sensor has the advantages of higher sensitivity, better specificity, accessibility, less sample consumption, and so forth. Moreover, it can be applied to accurately detect Pb2+ in complex biological or environmental samples, which is of great promise for widespread applications.

Monitoring of Parathion-Methyl Based on PtNPs Combined with 4-Amino-3-Hydroxy-1-Naphthalenesulfonic Acid Fluorescent Probe and Enzyme Inhibition

Monitoring of Parathion-Methyl Based on PtNPs Combined with 4-Amino-3-Hydroxy-1-Naphthalenesulfonic Acid Fluorescent Probe and Enzyme Inhibition

Micelle-Type Sensor for Saccharide Recognition by Using Boronic Acid Fluorescence Amphiphilic Probe and Surfactants

ABSTRACT Controlling molecular assembly by analytes is attractive for analytical techniques. Herein, we designed and synthesized amphiphilic boronic acid fluorescent probes (B-Nap-C8) possessing naphthaleneboronic acid as a saccharide recognition site and alkyl chain as a hydrophobic moiety. We evaluated their saccharide recognition function in relation to the fluorescent or absorption intensity changes of the assembled B-Nap-C8. Also, we evaluated the saccharide recognition function of B-Nap-C8 by the effect of co-surfactants. B-Nap-C8 forms micelles and recognizes fructose at pH 10. In the presence of surfactants, B-Nap-C8 with the cetyltrimethylammonium bromide (CTAB) micelle can recognize fructose at neutral pH. On the other hand, B-Nap-C8 with the sodium dodecyl sulfate (SDS) micelle recognizes fructose at pH > 10. This is caused by the decrease of the apparent pK a of B-Nap-C8 on the surface of the cationic CTAB micelle. The micelle formation of B-Nap-C8 with CTAB was confirmed by TEM observation, and the critical micelle concentration of B-Nap-C8 with CTAB was determined to be 1.79 mM from the CTAB titration measurements. From the saccharide titration experiments, B-Nap-C8 with CTAB micelles selectively recognizes fructose at neutral pH. The selectivity for saccharides was not observed in the saccharide titration of B-Nap-C8 with SDS. Thus, the co-surfactant systems with B-Nap-C8 were found to control the selectivity for saccharide recognition in water.

Inhibition of Lysozyme Amyloid Fibrillation by Silybin Diastereoisomers: The Effects of Stereochemistry.

Silybin is a flavonoid lignin compound consisting of two diastereomers with nearly equal molar ratios. It has been reported that silybin can effectively inhibit the aggregation of amyloid protein, but the difference between the two silybin diastereomers has been rarely studied. In this work, the inhibitory ability of silybin to hen egg-white lysozyme (HEWL) was demonstrated, and the difference of kinetic parameters of two diastereomers was analyzed. Fluorescence quenching titration was utilized to analyze the binding differences to native HEWL between the diastereomers, and transmission electron microscopy (TEM) was utilized to analyze the characteristics of the surface of various samples. The differences between hydrophobicity and the secondary structure among several HEWL samples were measured by the 8-anilino-1-naphthalene sulfonic (ANS) acid fluorescence probe, Raman spectra, and far-UV circular dichroism. Moreover, the differences in the binding energy of these two diastereomers with HEWL were analyzed by molecular docking. Also, we have investigated the effect of silybin diastereomers on HEWL fibril-induced cytotoxicity in SH-SY5Y cells. Results show that silybin has a certain inhibitory effect on the HEWL fibrillogenesis process, while silybin B (SB) has a more significant inhibitory effect than silybin A (SA), especially at high concentrations. This work provides some insights into the screening of amyloid inhibitors from complicated natural products and indicates that SB has the prospect of further development as an amyloid inhibitor.

A Cylindrical Ion Sensor Tip with a Diameter of 1.5 mm for Potentially Invasive Medical Application.

A fine cylindrical chemical sensor tip is developed with optical fiber in the core, surrounded by a transparent cylinder of photopolymer Norland Optical Adhesive 61 (NOA 61), and covered by a polymer hydrogel mixed with sensing molecules. The overall diameter is as small as 1.5 mm. pH response is demonstrated using two approaches of sensing materials: (i) absorbing probe Phenol Red mixed with Rhodamine 6G fluorescent dye and (ii) 8-hydroxypyrene-1,3,6-trisulfonic acid fluorescent probe. Both the optical excitation and fluorescence signal collection are through the optical fibers. A time resolution of 10 s is achieved for pH variations. Good linearity is observed in the physiological range from pH 7.0 to pH 8.6 with reversible and reproducible outcomes. For in vitro urea measurement, the sensor tip can distinguish 1, 3, and 5 mM urea solution, which is a crucial range in saliva urea concentration. The miniaturized tip with such simple cylindrical symmetry is designed to detect vital signs during minimally invasive surgeries and can be potentially accompanied with endoscopes to enter human bodies.

Photocatalytic Activity of Zn x Mn3-x O4 Oxides and ZnO Prepared From Spent Alkaline Batteries.

Oxides with ZnxMn3−xO4 stoichiometries and ZnO were synthesized from the “black mass” material recovered from spent alkaline batteries. The oxides were characterized by XRF, XRD with Rietveld refinement, SEM, and TEM methods. Optical characterization included diffuse reflectance (DRS) and photoluminescence (PL) measurements. ZnO presented a clear band edge in the UV region, and PL signals were detected. The Zn/Mn oxides showed strong absorption in the UV region and a continuous absorption band in the Vis-IR regions. There is a non-detected PL signal due to excited charges being trapped on sub-band energy states and/or transfer by non-radiative paths. Photocatalytic activity under both irradiation conditions was evaluated using the resazurin dye test, terephthalic acid fluorescence probe method, and NOx air purification evaluation. In the three photoactivity tests, ZnO performed well under both UV and Vis irradiation, whereas no evidence of any appreciable photocatalytic activity was observed for the Zn/Mn oxides. The results are discussed in terms of the findings of previously reported optical measurements.

Removal of antipyrine through two-dimensional and three-dimensional electrolysis: comparison, modification, and improvement

In this work, removal of antipyrine was studied through two-dimensional (2D) and three-dimensional (3D) electrolysis. 2D electrolysis was firstly studied with the Ti/SnO2-Ta2O5-IrO2 anode as working electrode. Operating parameters affecting antipyrine removal, such as current density, electrode distance, and initial concentration of antipyrine, were investigated and optimized. As the limited antipyrine removal efficiency of 48.0% was not satisfying, 3D electrolysis with γ-Al2O3 as particle electrodes was introduced in the purpose of improving the antipyrine removal. An obviously enhanced removal efficiency of 78.3% was obtained, which seemingly validated the effect of particle electrodes in improving antipyrine removal. Hence, an effort to further enhance the antipyrine removal efficiency was made through improving the electrochemical characteristics of γ-Al2O3 as particle electrodes. Modified Sn-Sb-Bi/γ-Al2O3 particles were thus prepared through impregnation method. And a desirable antipyrine removal efficiency of 94.4% and energy consumption of 0.18 kWh/g antipyrine were achieved in the 3D electrolysis with Sn-Sb-Bi/γ-Al2O3 as particle electrodes. Furthermore, possible mechanism and pathway of antipyrine degradation in 3D electrolysis were explored through detection of ·OH using terephthalic acid fluorescent probe method and detection of antipyrine degradation intermediates using LC-MS.

Design and Application of γ-aminobutyric Acid Nano-Fluorescent Probe

The design and development of nano-fluorescent probes is a research hotspot in the field of biological functional materials. The construction of γ-aminobutyric acid (GABA) fluorescent probe based on fluorescence resonance energy transfer (FRET) technology will become a new tool for bacterial screening. Recombinant plasmids suitable for prokaryotic expression were constructed by subcloning technology, and verified by agarose gel electrophoresis and sequencing. The target protein was expressed in BL21 E. coli and purified by nickel matrix affinity chromatography. Spectroscopy and fluorescence imaging validate probe function and evaluate bacteria’s environmental adaptability. The results showed that a recombinant plasmid suitable for efficient and stable expression of prokaryotes was successfully constructed. The release of GABA will cause changes in fluorescence intensity. The probe can evaluate the metabolic activity and adaptability of bacteria to different environments. This method uses bacteria’s metabolic activity to realize the selection of bacteria. The sample does not require special biochemical treatment. Compared with traditional evaluation methods by counting the number of bacteria and analyzing metabolite, it is a more convenient tool for screening and evaluation of bacteria strains.

Controllable synthesis of transition metal ion-doped CeO2 micro/nanostructures for improving photocatalytic performance

Ceria (CeO2) micro/nanostructures with various sizes and shapes were successfully synthesized by doping with a series of transition metal ions without any surfactant or template. The crystal structure, morphology, and photocatalytic performance of CeO2 and Co-, Zr-, Cu-doped CeO2 were characterised. The lattice parameters, which indicate the crystal structure distortion and change of Ce4+, were changed by incorporating a transition metal into the CeO2. Doping with transition metal ion could broaden the absorption range from ultraviolet to visible region and enhance the concentration of oxygen vacancies, which exhibited a significantly lower optical band than pure CeO2. The photocatalytic studies revealed that CeO2 with a spate of oxygen vacancies displayed a higher photocatalytic activity than pure CeO2 in degrading the organic pollutant rhodamine B (RhB). Furthermore, the O2− and OH radicals formed during photocatalysis process were revealed by means of nitrotetrazolium blue chloride (NBT) reduction method and a terephthalic acid (TA) fluorescence probe method, respectively, which discovers that radicals were crucial for the degradation of RhB. The H2-TPR confirmed that a small amount of transition metal ions significantly affected the oxidation state of the surface cations and oxygen vacancies. This study clearly reveals the effects of different transition metal ion dopants on the morphologies and photocatalytic performance of transition metal ion-doped CeO2 micro/nanostructures.

Synthesis and Verification of Fluorescent pH Probes Based on 2-Quinolone Platform

Intracellular pH plays an important role in biological events, including cell metabolism, signal transduction, cell growth, apoptosis and endocytosis. The development of simple and robust proton-recognizing fluorescent probes has been a research field of high interest. In this work, we describe the design and synthesis of a 2-quinolone Schiff base as a novel acidic fluorescent probe. The design strategy of the probe is based on non-classical hydrogen bonding in the form of an intramolecular three-centered hydrogen bond (THB). pH titrations at pHs 2 and 6 indicate that there is enhancement of fluorescence with increasing proton concentration. Additionally, no interference of metal ions such as Cu2+, Fe3+, Hg2+, Zn2+, Co2+, Cd2+ and Pb2+ ions were detected during the course of these titrations.

Responsive mechanism of three novel hypochlorous acid fluorescent probes and solvent effect on their sensing performance**Project supported by the National Natural Science Foundation of China (Grant Nos. 11374195 and 11404193) and the Taishan Scholar Program of Shandong Province, China.

Optical properties and responsive mechanisms of three newly synthesized fluorescent probes for hypochlorous acid (HOCl) are investigated by employing time-dependent density functional theory. The computational results show that the absorption and emission properties of these probes change obviously when they react with hypochlorous acid. It is found that the probe FHZ has the best performance according to the probing behavior. Moreover, the responsive mechanisms of the probes are studied by analyzing the distributions of molecular orbitals and charge transfer, which are shown as the photon-induced electron transfer (PET) for FHZ and the intramolecular charge transfer (ICT) for the other two probes. Specially, solvent effect on optical properties of the probe FHZ before and after reaction is studied within the polarizable continuum model (PCM). It is shown that performance of the probe depends crucially on the solvent polarity. Our computational results agree well with the experimental measurement, and provide information for design of efficient two-photon fluorescent probes.

A novel acidic pH fluorescent probe based on a benzothiazole derivative

A novel acidic pH fluorescent probe 1 based on a benzothiazole derivative has been designed, synthesized and developed. The linear response range covers the acidic pH range from 3.44 to 6.46, which is valuable for pH researches in acidic environment. The evaluated pKa value of the probe 1 is 4.23. The fluorescence enhancement of the studied probe 1 with an increase in hydrogen ions concentration is based on the hindering of enhanced photo-induced electron transfer (PET) process. Moreover, the pH sensor possesses a highly selective response to H+ in the presence of metal ions, anions and other bioactive small molecules which would be interfere with its fluorescent pH response. Furthermore, the probe 1 responds to acidic pH with short response time that was less than 1min. The probe 1 has been successfully applied to confocal fluorescence imaging in live HeLa cells and can selectively stain lysosomes. All of such good properties prove it can be used to monitoring pH fluctuations in acidic environment with high sensitivity, pH dependence and short response time.