Synthesis Of Fluorescent Probe Research Articles

A Novel NIR Fluorescence Probe with AIE Property to Image Viscosity in Nystatin-Induced Cell Model.

As one of the most significant parameters in cellular microenvironment, viscosity levels could be used to determine the metabolic process of bioactive substances within cells. Abnormal viscosity levels are closely associated with a series of diseases. Therefore, the design and synthesis of fluorescent probes that can monitor changes of intracellular viscosity in real-time is of great significance for the study of disease development process. Here, a new viscosity-recognized NIR fluorescence probe W1 based on quinoline-malonitrile is synthesized, and it is not susceptible to interference substances. Besides, AIE probe W1 shows fast response, excellent photostability, low cytotoxicity, good linear relationship between fluorescence intensity value and viscosity. Based on the above advantages, probe W1 is used to image the change of viscosity level in the cell model induced by nystatin.

Two-photon fluorescence probe for palladium with perchlorate induced quenching mechanism and its application in smartphone-based rapid detection

We propose a new approach for detecting palladium using a two-photon fluorescent probe quenched by perchlorate. This newly developed method has the potential to overcome some of the limitations of the currently available methods for detecting palladium. This article provides a detailed introduction to the design and synthesis of fluorescent probe, as well as the fluorescence performance in aqueous solutions. The results demonstrate the probe is highly sensitive, selective, and efficient in detecting palladium. The study also includes a thorough analysis of the quenching mechanism of the probe by perchlorate, and obtained different results from previous literatures. Moreover, the probe can easily identify and differentiate between palladium being present in the valence states 0, + 2/+ 4, and accomplish detecting palladium in convoluted solutions such as wastewater, environmental water, Hela cells and zebrafish. Due to its excellent performance, using self-developed optical device, the possibility of detecting palladium in aqueous solutions based on smartphone was explored.

Synthesis of fluorescent probe based on molecularly imprinted polymers on nitrogen-doped carbon dots for determination of tobramycin in milk.

Tobramycin (TOB) plays a considerable role in combating milk spoilage and preventing disease in dairy cows. However, overuse of TOB can lead to nephrotoxicity, ototoxicity, neuromuscular blockade, and hypersensitivity reactions. Here, nitrogen-doped carbon dots (N-CDs) were prepared with ethylenediamine and citric acid, then molecularly imprinted layers were obtained by imprinting of surface on the N-CDs to prepare Nitrogen-doped carbon dot-based molecularly imprinted polymers (N-CDs@MIPs). The fluorescence emission spectrum of this probe showed a linear enhancement with the TOB concentration in the 1-12μM. Meanwhile, a detection limit of 99.2nM was obtained. This probe was not affected by the structural analogs of the TOB and can show high sensitivity and selectivity compared to non-imprinted polymers (N-CDs@NIPs). Therefore, it can be successfully used for the trace analysis of TOB in milk with advantages over other reported techniques such as liquid chromatography coupled with tandem mass spectrometry or various aptamer sensors.

A novel benzo hemicyanine-based fluorescent probe for susceptible visualizing detection of phosgene

Leakage and misuse of phosgene, a common and highly hazardous industrial chemical, have always constituted a safety risk. Therefore, it is crucial to develop sensitive detection methods for gaseous phosgene. This work describes the design and development of a new fluorescent dye based on benzohemicyanine, as well as the synthesis of fluorescent probes for the sensitive detection of gaseous phosgene. Due to the excellent intramolecular charge transfer (ICT) effect from the strong electron-donating impact of the o-aminophenol group on benzo hemicyanine, the probe does not emit fluorescence. When the probe reacts with phosgene, the ICT effect is inhibited, and the result exhibits observable green fluorescence, thereby visualizing the response to phosgene. The probe offers exceptional sensitivity, a rapid response, and a low phosgene detection limit. In addition, we developed probe-loaded, portable test strips for the quick and sensitive detection of phosgene in the gas phase. Finally, the constructed probe-loaded test strips were utilized effectively to monitor the simulated phosgene leakage.

Synthesis of fluorescent probes based on cellulose for Fe2+ recognition

Synthesis of fluorescent probes based on cellulose for Fe2+ recognition

Discriminating normal and inflammatory mice models by viscosity changes with a two-photon fluorescent probe

Studies have found that the intracellular viscosity changes have close relationship with many diseases, therefore design and synthesis of fluorescent probe for testing intracellular viscosity is of great significance to the development of clinical. Herein, we developed a new two-photon near infrared probe (HCT) for viscosity imaging to discriminate normal and inflammatory models. Experimental results displayed that HCT has great sensitivity for the detection of viscosity, and based on the excellent performance of its photostability and lower cytotoxicity, HCT was successfully utilized for single-photon/ two-photon fluorescence imaging of the viscosity in living cells. More importantly, we employ HCT to further showcase in living tissues. Additionally, HCT could be used to discriminate between normal and inflamed mice, heralding its practical application in biomedical aspects.

Rational Design, Synthesis of Fluorescence Probes for Quantitative Detection of Amyloid-β in Alzheimer's Disease Based on Rhodamine-Metal Complex.

The efficient detection and monitoring of amyloid-β plaques (Aβ42) can greatly promote the diagnosis and therapy of Alzheimer's disease (AD). Fluorescence imaging is a promising method for this, but the accurate determination of Aβ42 still remains a challenge. The development of a reliable fluorescent probe to detect Aβ42 is essential. Herein, we report a rational design strategy for Aβ42 fluorescence probes based on rhodamine-copper complexes, Rho1-Cu-Rho4-Cu, among them Rho4-Cu exhibits the best performance including high sensitivity (detection limit = 24 nM), high affinity (Kd = 23.4 nM), and high selectivity; hence, Rho4-Cu is selected for imaging Aβ42 in AD mice, and the results showed that this probe can differentiate normal mice and AD mice effectively.

Precise probe design based ESIPT coupled AIE mechanism toward endogenous cyanide in food detection and bioimaging

Cyanide (CN–) is one of the strongest and fastest-acting toxic substances, and its excessive presence will cause great harm to the environment, food and human health. Although several fluorescent probes that recognize CN– have been extensively reported, developing the fluorescent probe with aggregation-induced emission (AIE) and large Stokes shift is still an extremely difficult problem due to lack of effective strategies. Thus, continuing efforts to seek novel, structurally distinct and functionally specific structures with large Stokes shift and AIE features are highly anticipated. Herein, we design and synthesize a novel asymmetric bis-Schiff base block (named as HPBC) based on the excited state intramolecular proton transfer (ESIPT) and AIE mechanism. Benefiting from restriction of intramolecular rotation and –CN electron-withdraw nature promoting the ESIPT effect of HPBC in the recognition system, HPBC shows large Stokes shift (243 nm), high selectivity, rapid identification of CN– and AIE effects. Excitingly, the limit of detection (LOD) was calculated to be 1.32 × 10-7 M, which was much lower than the 1.9 × 10-6 M specified by the World Health Organization and better than other previously reported probes. More importantly, further investigations demonstrated that the molecule we designed was successfully applied to sense endogenous CN– in food samples and the low cytotoxicity of HPBC was allowed to discover CN– in C. elegans, mice and HeLa cells. Overall, this work provides a new viewpoint of the rational design and synthesis of fluorescent probe toward CN– with AIE and large Stokes shift features, and triggers the discovery of new functions and properties of biological materials, which would open a new frontier for probes.

Molecular engineering of an efficient iminocoumarin-based probe for practical sensing applications

Thiophenols are important synthetic intermediates that are also primary pollutants. So, it is of great importance to develop highly sensitive and selective approaches for the detection of thiophenols in practical samples and living systems. Herein, an efficacious high-throughput strategy (benzopyran ring-opening reaction with iminocoumarins and alkylating reagents), was developed for the synthesis of fluorescent probes. The selected thiophenol-specific probe IminoC1 showed remarkable sensing performance with a 320-fold fluorescence enhancement, good detection limit of 31 nM, and high selectivity over common biothiols. Moreover, the probe IminoC1 was successfully applied for the determination of thiophenols in water samples and the visualization of thiophenols in living cells, which pointed to its strong potential for practical sensing applications.

Specific fluorescent probe synthesis based on “thiol-chromene click reaction” and its applications

Specific fluorescent probe synthesis based on “thiol-chromene click reaction” and its applications

Synthesis of Fluorescent Probes Targeting Tumor-Suppressor Protein FHIT and Identification of Apoptosis-Inducing FHIT Inhibitors.

For the early diagnosis of cancer, leading to a better chance of full recovery, marker genes whose expression is already altered in precancerous lesions are desirable, and the tumor-suppressor gene FHIT is one candidate. The gene product, FHIT protein, has a unique dinucleoside triphosphate hydrolase (AP3Aase) activity, and in this study, we designed and synthesized a series of FHIT fluorescent probes utilizing this activity. We optimized the probe structure for high and specific reactivity with FHIT and applied the optimized probe in a screening assay for FHIT inhibitors. Screening of a compound library with this assay identified several hits. Structural development of a hit compound afforded potent FHIT inhibitors. These inhibitors induce apoptosis in FHIT-expressing cancers via caspase activation. Our results support the idea that FHIT binders, no matter whether inhibitors or agonists of AP3Aase activity, might be promising anticancer agents.

Fluorescence Intensity Enhancement of Green Carbon Dots: Synthesis, Characterization and Cell Imaging.

The hydrothermal treatment of green carbon dots (CDs) is an appropriate fluorescent probe synthesis method. CDs are exploited as biological staining agents, especially for cellular detection and imaging. The nitrogen-doped green carbon dots (N-CDs) formation can improve the fluorescence intensity property in a one-step process. Here, we report two N-CDs from lemon and tomato extraction in the presence of hydroxylamine. Lemon and tomato N-CDs showed the blue fluorescence under ultraviolet radiation of about 360nm. The characterization of CDs and N-CDs showed the presence of N-H and C-N bonds which enhanced the fluorescence efficiency. The mean size of lemon and tomato N-CDs were about 2 and 3nm with an increased quantum yield (QY) of 5% and 3.38%, respectively. The CDs and N-CDs cytotoxicity assay exhibited high cell viability approximately 85% and 73%, respectively. N-CDs show superior fluorescent intensity in different solvents and significant stability under long-time UV irradiation, different PH and high ionic strength. Our results indicated that the use of N-CDs in cell imaging can lead to fluorescence intensity enhancement as well as proper biocompatibility. Therefore, the safe and high fluorescence intensity of green N-CDs can be utilized for fluorescent probes in biolabeling and bioimaging applications.

Design, synthesis and evaluation of a novel fluorescent probe to accurately detect H2S in hepatocytes and natural waters

Design and synthesis of fluorescent probe with fast response, excellent water solubility and good hepatocyte-targeting capacity to detect hydrogen sulfide (H2S) in hepatocytes and water samples is of great significance. Here, a novel fluorescent probe QL-Gal-N3 for detection of H2S was designed and synthesized based on H2S-mediated azide reduction strategy. This sensor demonstrated low toxicity, fast response (within 1 min), high selectivity and low detection limit (as low as 126 nM in water) for the detection of H2S. HeLa, A549 and HepG-2 cells were chosen to investigate the hepatocyte-targeting ability of QL-Gal-N3 respectively. The results indicated that the specific recognition of ASGPR over-expressed in hepatocytes by galactose group was an important reason for the good targeting ability of probe QL-Gal-N3. Furthermore, due to the introduction of glycosyl moiety, the water solubility of fluorescent probe was enhanced obviously. It was successfully applied for the detection of H2S in environmental water samples including river water, tap water, lake water and waste water.

Spermine-NBD as fluorescent probe for studies of the polyamine transport system in Leishmania donovani.

This study describes the synthesis of fluorescent probes as potential substrates for the polyamine transport system (PTS) of Leishmania donovani. A competitive radioassay was used to determine the most efficient probe. We observed that the conjugate spermine-nitrobenzofurazan (Spm-NBD) was able to compete with [3H]-spermidine in L. donovani at a potent IC50 of 60 µM.

Chemoselective Acylation of Hydrazinopeptides to Access Fluorescent Probes for Time-Resolved FRET Assays on GPCRs.

Fluorescence techniques represent a powerful tool to investigate dynamic and functional architecture of GPCRs. Thus, fluorescent GPCR ligands have found various applications in cellular imaging, in the development of binding assays as replacements for radioligands in the study of ligand-receptor but also in receptor-receptor interactions at the cell surface or in native tissues. To extend the applicability of these techniques, the design and the synthesis of fluorescent probes are critical steps. As there are numerous peptide receptors in the GPCR family, fluorescent peptide-based probes are of importance. Herein, we present a convenient method to facilitate the solution-phase fluorescent labeling of peptides which is based on the chemoselective acylation of α-hydrazinopeptides. This approach combines the advantages to use commercially available amine-reactive dyes and very mild conditions that are fully compatible with the chemical sensitivity of the dyes. It gives a rapid access to fluorescent peptidic probes compatible with the time-resolved fluorescence resonance energy transfer (TR-FRET) techniques.

Synthesis and Structure-Activity Relationship of Omaezallene Derivatives.

Omaezallene derivatives (nor-bromoallene, nor-bromodiene, and bromoenynes) were successfully synthesized. Their antifouling activity and toxicity to the cypris larvae of the barnacle Amphibalanus amphitrite and ecotoxicity to the marine crustacean Tigriopus japonicus were studied. It was revealed that the two side chains of omaezallene were essential to its antifouling activity because the activities of nor-bromoallene and nor-bromodiene were significantly diminished. The bromoenyne was found to exhibit potent antifouling activities comparable to omaezallene with low toxicity and ecotoxicity. Preparation of bromoenyne framework is much easier than that of bromodiene moiety in omaezallene. Based on the antifouling activities of the bromoenynes, the synthesis of fluorescent probes and evaluation of their biological activities were also carried out.

Sensing Performance and Efficiency of Two Energy Transfer-Based Two-Photon Fluorescent Probes for pH.

The design and synthesis of fluorescent probes for monitoring pH values inside living cells have attracted great attention, due to the important role pH plays in many biological processes. In this study, the optical properties of two different two-photon fluorescent probes for pH are studied. The ratiometric sensing of the probes are theoretically illustrated. Meanwhile, the recognitional mechanisms of the probes are investigated, which shows the energy transfer process when react with H+. Specially, the calculated results demonstrate that Probe1 possesses a higher energy transfer efficiency and a larger two-photon absorption cross-section than Probe2, indicating it to be a preferable pH fluorescent probe. Therefore, the influence of connection between the donor and the acceptor on the sensing performances of the probe is demonstrated. Our results help to understand the experimental observations and provide a theoretical basis to synthesize efficient two-photon fluorescent probes for monitoring pH changes.

Heterocyclic sterol probes for live monitoring of sterol trafficking and lysosomal storage disorders

The monitoring of intracellular cholesterol homeostasis and trafficking is of great importance because their imbalance leads to many pathologies. Reliable tools for cholesterol detection are in demand. This study presents the design and synthesis of fluorescent probes for cholesterol recognition and demonstrates their selectivity by a variety of methods. The construction of dedicated library of 14 probes was based on heterocyclic (pyridine)-sterol derivatives with various attached fluorophores. The most promising probe, a P1-BODIPY conjugate FP-5, was analysed in detail and showed an intensive labelling of cellular membranes followed by intracellular redistribution into various cholesterol rich organelles and vesicles. FP-5 displayed a stronger signal, with faster kinetics, than the commercial TF-Chol probe. In addition, cells with pharmacologically disrupted cholesterol transport, or with a genetic mutation of cholesterol transporting protein NPC1, exhibited strong and fast FP-5 signal in the endo/lysosomal compartment, co-localizing with filipin staining of cholesterol. Hence, FP-5 has high potential as a new probe for monitoring cholesterol trafficking and its disorders.

Dipodal quinoline-tethered fluorescent probe synthesis and investigation of spectroscopic properties

Novel quinoline-tethered fluorescent probe was designed and synthesized as multidentate ligand. Their sensing actions were confirmed by UV–Vis absorbance and emission spectroscopic studies in presence of perchlorate salts of Co2+, Li+, Fe2+, K+, Pb2+, Cu2+, Zn2+, Ni2+, Hg2+, Ag+ cations in acetonitrile (1 × 10−5 M for absorption studies, 1 × 10−7 M for fluorescence studies). It was found that the dipodal compounds can selectively bind to Cu2+ and Ag+ metal ions with a significant quenching in their emissions. The capture of Cu2+ and Ag+ by the probe resulted in deprotonation of the secondary amine conjugated to the quinoline-tethered probe, so that the electron-donation ability of the ‘N’ atom would be greatly enhanced and the probe (2) showed blue-shift in emission and exhibited an on-off fluorescent response. The binding study was explored by using fluorescence spectroscopy with Job plot method.

Real-Time Imaging of the Azole Class of Antifungal Drugs in Live Candida Cells.

Azoles are the most commonly used class of antifungal drugs, yet where they localize within fungal cells and how they are imported remain poorly understood. Azole antifungals target lanosterol 14α-demethylase, a cytochrome P450, encoded by ERG11 in Candida albicans, the most prevalent fungal pathogen. We report the synthesis of fluorescent probes that permit visualization of antifungal azoles within live cells. Probe 1 is a dansyl dye-conjugated azole, and probe 2 is a Cy5-conjugated azole. Docking computations indicated that each of the probes can occupy the active site of the target cytochrome P450. Like the azole drug fluconazole, probe 1 is not effective against a mutant that lacks the target cytochrome P450. In contrast, the azole drug ketoconazole and probe 2 retained some antifungal activity against mutants lacking the target cytochrome P450, implying that both act against more than one target. Both fluorescent azole probes colocalized with the mitochondria, as determined by fluorescence microscopy with MitoTracker dye. Thus, these fluorescent probes are useful molecular tools that can lead to detailed information about the activity and localization of the important azole class of antifungal drugs.