Probe For Detection Research Articles

An AuNPs/Co3O4 nanocubic cage–based electrochemiluminescent immunosensor for Corin: a biomarker of stroke

Stroke, or cerebral apoplexy, is a paroxysmal disease that seriously threatens the neurological functions of middle-aged and older men, resulting in high rates of disability and mortality. The blood level of Corin can accurately predict the risk and subtype of stroke. Herein, an immunosensor was developed utilizing a nanocomposite of Au nanoparticles and Co3O4 nanocubic cages (AuNPs/Co3O4NcC) as the sensing substrate, with luminol serving as the sensing probe for Corin detection. Through the application of 3-aminopropyltrimethoxysilane, AuNPs/Co3O4NcC was affixed to the surface of indium tin oxide (ITO)-coated glass, considerably enhancing the electrochemiluminescence (ECL) performance of luminol due to its peroxidase-like activity. To confer biological specificity by immobilizing the Corin antibody on this substrate and to block residual active sites with bovine serum albumin, an immunosensor was developed. The capture of Corin protein by the antibody through an immune reaction, leading to its binding on the sensor, resulted in a decreased ECL signal. Under optimized conditions, the proposed ECL immunosensor demonstrated a broad linear response range from 1pg/mL to 4800pg/mL and a low detection limit of 0.26pg/mL. This ECL immunosensor is characterized by its specificity, stability, and reproducibility and can detect Corin in real blood samples.

Near-infrared fluorescent probe for selective and sensitive detection of glutathione based on thioctic acid-functionalized Ag/Au NCs-assisted by ferric ion

Accurate determination of glutathione (GSH) levels in biological fluids and food supplements is paramount. To address this need, we developed a near-infrared (NIR) fluorescent probe with high sensitivity and selectivity for GSH detection. This probe comprises thioctic acid-functionalized silver and gold nanoclusters (ThA@Ag/Au NCs), emitting red light at 720 nm upon excitation at 310 nm. In the presence of Fe3+ ions, which form complexes with ThA, the fluorescence of ThA@Ag/Au NCs is quenched due to aggregation. However, upon addition of GSH, a potent chelator for Fe3+, the fluorescence of the probe is restored, leading to enhanced emission. Under optimized conditions, the fluorescence response (F/F°) exhibits a linear relationship with increasing GSH concentration within the range of 0.03–95 µM, achieving a limit of detection (LOD) of 9.0 nM. This designed scheme demonstrates excellent selectivity, sensitivity, and accuracy for GSH detection. Furthermore, the NIR fluorescent probe effectively estimated GSH levels in various matrices, including capsules, serum, urine, and saliva samples, yielding recoveries % ranging from 97.2 % to 103.2 %, with RSD % below 4.34 %. Due to its NIR properties, this fluorescent probe holds promise for diagnosing various diseases and potential in vivo applications.

An experimental and theoretical study on mechanistic insights into urolithin-based ratiometric fluorescent probe for instant quantitative detection of fluoride ions

Fluoride detection has been playing an important role in chemical, biological, and medicinal field, especially for keeping physical health and resisting environmental pollution. Herein, a urolithin B fluorescent probe has been successfully developed with good sensitivity, selectivity, anti-interference ability. The low limit of detection (LOD) refers to 0.156 μM, and the instant response time to F− is less than 1 s. The probe is suitable for quantitatively and qualitatively ratiometric detection for F− in solution with two distinct emission bands at 425 (blue) and 566 nm (orange), with the coordinate change of CIE from (0.38, 0.41) to (0.22, 0.11). Urolithin B displayed a remarkable ratiometric fluorescence response towards F−. The detection mechanistic was further proposed by NMR and electronic spectroscopic experiments combining with time-dependent density functional theoretical calculation.

Si,N co-doped carbon quantum dots in mesoporous molecular sieves: A fluorescence sensing platform for Cr(VI) detection

Carbon quantum dots (CQDs) are highly efficient fluorescent probes for metal ion detection; however, they are limited by non-uniform particle size, aggregation, and the inability to facilitate quantitative analysis. Herein, a smart host–guest strategy is utilized, wherein CQDs are immobilized in mesoporous molecular sieves, proving to be an effective method to minimize these drawbacks. Si,N-CQDs@SBA-15 is successfully synthesized by the one-step hydrothermal route using Si,N-CQDs precursor materials as guests and the mesoporous molecular sieve SBA-15 as the host. A good linear relationship between the concentration of Cr(VI) ions and fluorescent intensity is observed from 0 μM to 100 μM (R2 = 0.9986). Thelimitofdetection(LOD) is 0.213 μM, approximately three times lower than that of pristine Si,N-CQDs (0.677 μM). Additionally, Si,N-CQDs@SBA-15 exhibit good detection selectivity for Cr(VI) detection over other metal ions. Moreover, a smartphone-based laboratory device and RGB analysis app are employed to capture and analyze fluorescence images, revealing a LOD for Cr(VI) of 0.201 μM. Förster Resonance Energy Transfer (FRET) is identified as the possible mechanism for the fluorescence quenching of Si,N-CQDs@SBA-15 by Cr(VI) through UV–vis and fluorescence experiments. The practicability of the synthesized fluorescent probes is further validated in river water, yielding satisfactory spiked recoveries ranging from 99.24 % to 106.21 %.

Carbon quantum dots as a turn-on fluorescent probe for the sensitive detection of Cd2+

In this study, fluorescent carbon dots were synthesized for the first time using ammonium citrate and glutamic acid as precursors via a one-pot hydrothermal method. The synthesized carbon dots emit blue fluorescence at 436 nm (excited at 320 nm) and demonstrate excellent photobleaching resistance and fluorescence stability in high salt environments. Within the range of 1–25 μM, the fluorescence of CDs gradually increases with the increasing concentration of Cd2+, reaching a limit of detection as low as 13 nM. This phenomenon could be ascribed to the chelation-enhanced fluorescence, a result of Cd2+ forming complexes with the abundant surface functional groups such as CN–, –COOH, –OH, –NH2 in CDs. Furthermore, this turn-on fluorescent probe has been successfully used for the detection of Cd2+ in tap water and lake water, providing an efficient and sensitive method for the analysis of environmental metals.

A dual-ratiometric strategy to design the NIR-IIb reference-based activatable nanoprobe for accurate nitric oxide detection

Fluorescence imaging offers a real-time, convenient method for detection nitric oxide (NO) in vivo, especially using the near-infrared IIb (NIR-IIb, 1500-1700nm) region. However, traditional single-ratiometric fluorescent probes for NO detection suffer from signal distortion due to inherent limitations such as signal crosstalk and component separation. Herein, we proposed a dual-ratiometric strategy to ensure detection accuracy. The detection result was considered to be reliable upon the auxiliary ratio and practical ratio exhibiting a synchronous change. Based on this strategy, we fabricated a NO-responsive probe, comprising a conjugated polymer P-NH2 and a rare-earth doped nanocrystal NaYF4:Yb,Er,Ce@NaYF4:Yb,Nd@NaYF4. When interaction with NO, the probe generated two ratiometric signals: FNIR-I/FNIR-IIb and FNIR-II’/FNIR-IIb (NIR-I, 700-900nm; NIR-II’, 1000-1400nm), where the latter acted as the practical ratio for its better imaging clarity. With an acetaminophen-induced liver injury model, the probe demonstrated the potential for precise ratiometric imaging of NO distribution in vivo. Particularly, ratio imaging revealed the intricate vascular structure of the abdomen, demonstrating the significant advantages of the NIR-IIb region. Our study offers an effective approach to reduce detection errors in the bioimaging applications and provides a powerful tool for basic biomedical research.

Visual detection and discrimination of nerve and blood agents using a dual-site fluorescent probe in living cells and mice

Of all chemical warfare agents (CWAs), only nerve and blood agents cause massive mortality at low concentrations. To better detect and discriminate nerve and blood agents, a reliable detection method is desirable. We report a series of fluorescent probes for nerve and blood agent detection. Among the tested probes, SR-Pip detected nerve and blood agents quickly (within 10s for nerve agents and 1min for blood agents). SR-Pip coupled with nerve agent produced a weak orange fluorescence with good sensitivity [limit of detection (LOD)=5.5μM]. Upon reaction with blood agent, the fluorescence of SR-Pip changed from orange fluorescence to blue fluorescence with detection limits as low as 9.6nM. This probe effectively visualised different concentrations of nerve agents in living cells, mice and blood. A portable test kit using SR-Pip instantly detected nerve and blood agents. To the best of our knowledge, SR-Pip is the first fluorescent probe for nerve and blood agent detection. Environmental ImplicationBlood agents (also known as cyanide) are widely used in industrial chemicals. Accidental spills of cyanide solutions into rivers and streams have produced massive kills of fish and other aquatic biota. In addition, cyanide intoxication can occur under the conditions of terrorist activities, smoke inhalation, and even improperly prepared foods. Nerve agents are seldom used in industrial chemicals, but they are a class of highly toxic organophosphate compounds. Nerve agents remain a clear and present danger to both the military and citizens. Therefore, it is important to develop a convenient, accurate detection method for nerve and blood agents.

Intermolecular π-π interactions-induced aggregation based on bispyrene for ultrasensitive detection of picric acid (PA) and 2,4-dinitrophenol (DNP)

It was still important to explore excimer emission-based fluorescence probes for nitroaromatics detection by formation or destruction of pyrene excimer with strong emission change. In this work, we have synthesized a simple bis(pyrene) derivative (BisPyH) and studied its optical behavior by using UV–vis and fluorescent spectroscopy. In pure DMSO solution, it emitted a weak blue fluorescence due to the formation of excimer states, on the contrary, the BisPyH in the DMSO/H2O (v/v, 1:1) mixture solution was self-assembled to form aggregates via intermolecular π-π interactions among pyrene units, giving almost 10-fold fluorescence enhancement compared to that in pure DMSO solution. The formed aggregates just offered a perfect platform for the selective detection PA and DNP explosives by quenching the bright green intermolecular-excimer emission in competitive environment. BisPyH exhibited a limit of detection (LOD) towards PA, and DNP as low as 7.32 × 10−8, and 8.11 × 10−8 M, respectively. Subsequently, the fluorescence quenching mechanism was demonstrated by the density functional theory (DFT), UV–vis and fluorescence spectra, SEM, and the quenching constants. Furthermore, filter paper strips and thin-layer chromatography (TLC) plates coated with BisPyH were able to detect PA and DNP in the practical application.

Porphyrin-based covalent organic framework as oxidase mimic for highly sensitive colorimetric detection of pesticides.

A covalent organic framework-based strategy was designedfor label-free colorimetric detection of pesticides. Covalent organic framework-based nanoenzyme with excellent oxidase-like catalytic activity was synthesized. Unlike other artificial enzymes, porphyrin-based covalent organic framework (p-COF) as the oxidase mimic showed highly catalytic chromogenic activity and good affinity toward TMB without the presence of H2O2, which can be used as substitute for peroxidase mimics and H2O2 system in the colorimetric reaction. Based on the fact that the pesticide-aptamer complex can inhibit the oxidase activity of p-COF and reduced the absorbance at 650nm in UV-Vis spectrum, a label-free and facile colorimetric detection of pesticides was designed and fabricated. Under the optimized conditions, the COF-based colorimetric probe for pesticide detection displayed high sensitivity and selectivity. Taking fipronil for example the limit of detection was 2.7ng/mL and the linear range was 5-500,000ng/mL. The strategy was successfully applied to the detection of pesticides with good recovery , which was in accordance with that of HPLC-MS/MS. The COF-based colorimetric detection was free of complicated modification H2O2, which guaranteed the accuracy and reliability of measurements. The COF-based sensing strategy is a potential candidate for the sensitive detection of pesticides of interests.

SMS Fiber Structure-Based Enzymatic Sensor Probe for Uric Acid Detection Using Gold Nanoparticles and Graphene Oxide

SMS Fiber Structure-Based Enzymatic Sensor Probe for Uric Acid Detection Using Gold Nanoparticles and Graphene Oxide

A new fluorescent probe for detection of hydrazine and extremely acidic pH in different modes

Toxic hydrazine poses a huge threat to living organisms and our environment. Extremely acidic pH is also harmful to various ecosystems. Here, a coumarin-based dye called CMM has been synthesized for the ratiometric detection of hydrazine (F504 nm/F560 nm). CMM could detect gaseous hydrazine in the environment and endogenous hydrazine in live cells. Furthermore, the fluorescence at 563 nm of CMM was stable from pH 10.0 to 2.0, whereas increased greatly from pH 2.0 to 1.2. Thus, the probe could also detect extremely acidic pH with great fluorescence enhancement. The pKa value was determined to be as low as 1.41. We anticipate that the probe could provide a good candidate for detection of toxic hydrazine and extremely acidic pH.

A hemicyanine-based fluorescent probe for detection of pH and its applications in live cells imaging and food monitoring

pH as an important parameter is closely related to many pathological processes and food freshness. In this paper, a new pH-sensitive fluorescent probe 1 based on hemicyanine dye was synthesized and used for the detection of pH in live cells and food spoilage. Under neutral and alkaline conditions, the phenolic hydroxyl group of the probe deprotonates, forming a closed spirocyclic structure and resulting in fluorescence quenching. Under acidic conditions, the probe maintains an open-loop structure and has intense fluorescence. The probe possessing good selectivity and stability can target lysosomes and visualize the exogenous pH in live cells. Furthermore, by using this probe, we successfully achieved the monitoring of pH in metamorphic process of shrimp and pork. This probe is expected to provide a good imaging tool for real-time detection of pH in live cells and food monitoring.

Bacterial lipase-responsive polydopamine nanoparticles for detection and synergistic therapy of wound biofilms infection

Wound biofilms represent an elusive conundrum in contemporary treatment and diagnostic options, accredited to their escalating antibiotic resistance and interference in chronic wound healing processes. Here, we developed mesoporous polydopamine (mPDA) nanoparticles, and grafted with rhodamine B (Rb) as biofilm lipase responsive detection probe, followed by π − π stacking mediated ciprofloxacin (CIP) loading to create mP-Rb@CIP nanoparticles. mPDA NPs with a melanin structure could quench fluorescence emissions of Rb. Once encountering biofilm in vivo, the ester bond in Rb and mPDA is hydrolyzed by elevated lipase concentrations, triggering the liberation of Rb and restore fluorescence emissions to achieve real-time imaging of biofilm-infected wounds. Afterwards, the 808 nm near-infrared (NIR) illumination initiates a spatiotemporal controlled antibacterial photothermal therapy (PTT), boosting its effectiveness through photothermal-triggered CIP release for synergistic biofilm eradication. The mP-Rb@CIP platform exhibits dual diagnostic and therapeutic functions, efficaciously treating biofilm-infected wounds in vivo and in vitro. Particularly, the mP-Rb@CIP/NIR procedure expedites wound-healing by alleviating oxidative stress, modulating inflammatory mediators, boosting collagen synthesis, and promoting angiogenesis. Taken together, the theranostic nanosystem strategy holds significant potential for addressing wound biofilm-associated infections.

A near-infrared turn on fluorescent probe for specific detection of cysteine and its application based on a corrole fluorophore

A near-infrared turn on fluorescent probe for specific detection of cysteine and its application based on a corrole fluorophore

Zirconium-porphyrin-MOF-based oxidase-like nanozyme with oxygen vacancy for aflatoxin B1 colorimetric sensing.

In this study, a porous coordination network zirconium-porphyrin-based nanoparticle with oxygen vacancies (OVs) was prepared using acetic acid and benzoic acid as modulators via a simple hydrothermal method. The presence of OVs was confirmed by various characterization methods and was found to enhance oxygen uptake and activation. This resulted in the generation of more reactive peroxyl radicals (•O2 -) and led to an improved oxidase (OXD) mimetic activity. Additionally, it promoted 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) oxidation, with a low Km value of 0.07mM and a high Vmax of 1.47×10-7M·s-1. As aflatoxin B1 (AFB1) inhibits the Pt@PCN-222-ABTS nanozyme system, a colorimetric probe for AFB1 detection was constructed. The limit of detection (LOD)was 0.074µg·L-1. This research presents a novel approach for designing a nanozymatic-based colorimetric method to analyze trace AFB1 residues in food.

Genetic Encoding of Fluoro-l-tryptophans for Site-Specific Detection of Conformational Heterogeneity in Proteins by NMR Spectroscopy.

The substitution of a single hydrogen atom in a protein by fluorine yields a site-specific probe for sensitive detection by 19F nuclear magnetic resonance (NMR) spectroscopy, where the absence of background signal from the protein facilitates the detection of minor conformational species. We developed genetic encoding systems for the site-selective incorporation of 4-fluorotryptophan, 5-fluorotryptophan, 6-fluorotryptophan, and 7-fluorotryptophan in response to an amber stop codon and used them to investigate conformational heterogeneity in a designed amino acid binding protein and in flaviviral NS2B-NS3 proteases. These proteases have been shown to present variable conformations in X-ray crystal structures, including flips of the indole side chains of tryptophan residues. The 19F NMR spectra of different fluorotryptophan isomers installed at the conserved site of Trp83 indicate that the indole ring flip is common in flaviviral NS2B-NS3 proteases in the apo state and suppressed by an active-site inhibitor.

AIEE-active dichlorobenzene and chlorobenzene ratiometric fluorescent probe based on [2.2]paracyclophane

Two AIEE-active [2.2]paracyclophanyl-based diester and monoester (1a and 1b) were facilely synthesized by one-pot method and applied as ratiometric fluorescent probe to detect dichlorobenzene (DCB) and chlorobenzene (CB). Compared with compound 1b, 1a exhibits high sensitivity and low detection limits for DCB and CB in dichloromethane (DCM), particularly, the detection sensitivities for ortho-dichlorobenzene (o-DCB), meta-dichlorobenzene (m-DCB) and chlorobenzene can be modulated by AIEE behavior with lower detection limits of 23.64, 56.27, and 5.92 ppm, respectively in THF/H2O mixed solutions with water fraction (fw) of 70 % due to the formation of aggregation-state. The X-ray structure analysis, theoretical calculations and photophysical properties in different solvents were investigated to reveal the distinctive photophysical behaviors of 1a and 1b. The facile synthesis, X-ray structure, AIEE modulated sensing properties for o-DCB, m-DCB, and CB in DCM and THF/H2O mixed solutions make 1a potential application as fluorescent probe for trace DCB and CB detection in drinking water.

MOF-based probes for total content detection of Ca2+/Sr2+----Regulating detection performance indicators of probes via "paramagnetic/diamagnetic heterometallic probes" strategy

Probes based on luminescence metal–organic frameworks (LMOFs) have been extensively investigated. Although many of the probes can detect multiple analytes individually, it has not been reported that the probes can detect the total content of multiple analytes simultaneously. Herein a series of isostructural MOFs ([MgxMn1-x(L1)(DMSO)2]) were constructed via a strategy utilizing paramagnetic/diamagnetic heterometallic probes, which can be used for detecting Ca2+/Sr2+ individually or simultaneously without discrimination through a structural transformation mechanism. In-depth investigations reveal that probes involving such mechanism possess distinct advantages in total content detection, and one noteworthy characteristic of the probes is they exhibit similar working curves for different detectable analytes. By adjusting the ratio of paramagnetic/diamagnetic metal ions in the probes, their detection performance indicators can be precisely regulated. This work not only develops a new strategy for the customization of probe with specific performance indicators, but also opens up fresh opportunities to construct probes for total content detection.

A sensitive analytical strategy of oligonucleotide functionalized fluorescent probes for detection of nusinersen sodium in human serum

Spinal muscular atrophy (SMA) is a rare autosomal recessive neuromuscular disease. Nusinersen sodium (NS) is the world's first antisense oligonucleotide (ASO) drug for SMA precise targeted therapy. However, the limited half-life of oligonucleotides and their tendency to accumulate in hepatic and renal tissues presented significant challenges for clinical investigation and therapeutic drug monitoring. In this study, we proposed an analytical strategy based on the specific capture of oligonucleotide functionalized fluorescent probes by single stranded binding proteins (SSB) for ultra-sensitive and high-throughput detection of nusinersen sodium in human serum. The magnetic nanoparticles modified with single-strand binding protein (MNPs-SSB) selectively bonded to the red fluorescent quantum dots functionalized with oligonucleotides (RQDs-ssDNA) that were complementary to nusinersen sodium. Upon interaction with nusinersen sodium, RQDs-ssDNA formed a double-stranded complex (RQDs-ssDNA-NS), resulting in enhanced red fluorescence after magnetic separation as it was no longer captured by MNPs-SSB but remained in the supernatant. A quantitative analysis of nusinersen sodium in biological samples was successfully achieved by establishing a relationship between fluorescence intensity and its concentration. The detection signal F/F0 exhibited a linear correlation (R2 = 0.9871) over a wide range from 0.1 nM to 200 nM, with a limit of detection (LOD) of 0.03 nM, demonstrating the high specificity and rapid analysis time (only 30 min). This method provided a novel approach for sensitive, high-throughput, and specific analysis of nusinersen sodium and similar ASO drugs.

Advancements in Small Molecule Fluorescent Probes for Superoxide Anion Detection: A Review.

Superoxide anion (O2•-), a significant reactive oxygen species (ROS) within biological systems, plays a widespread role in cellular function regulation and is closely linked to the onset and progression of numerous diseases. To unveil the pathological implications of O2•- in these diseases, the development of effective monitoring techniques within biological systems is imperative. Small molecule fluorescent probes have garnered considerable attention due to their advantages: simplicity in operation, heightened sensitivity, exceptional selectivity, and direct applicability in monitoring living cells, tissues, and animals. In the past few years, few reports have focused on small molecule fluorescence probes for the detection of O2•-. In this small review, we systematically summarize the design and application of O2•- responsive small molecule fluorescent probes. In addition, we present the limitations of the current detection of O2•- and suggest the construction of new fluorescent imaging probes to indicate O2•- in living cells and in vivo.