Organic Fluorescent Probes Research Articles

Fluorescence imaging of dopamine in living cells triggered by unique coupling reactions

BackgroundDopamine (DA) is a significant neurotransmitter and catecholamine molecule in the mammalian brain. It plays a crucial role in perception, cognition, central nervous system regulation, and hormone secretion. The detection of DA levels in living systems is a vital aspect of the research and early diagnosis of neurological disorders. ResultsIn this study, we achieved an effective fluorescence generation specific to DA under physiological pH conditions by coupling the DA detection reaction to the HClO oxidation reaction. This method has been used for the detection of DA in cells, as well as for the visualization of DA levels in cellular models of inflammation and depression. SignificanceThis is the first time that utilizes an organic fluorescent molecular probe to detect DA in living cells.

A multifunctional metal–organic complex fluorescent probe for highly sensitive detection of lysine, CrO42-/Cr2O72-, Fe3+ and nitro-aromatic compounds

Water contamination caused by organic and inorganic compounds represents an urgent global issue. It is meaningful to detect these compounds. A metal–organic complex (Zn-HTBA) was investigated as a multifunctional fluorescent probe which showed exceptional fluorescence characteristics. The Zn- HTBA can specifically detect lysine in water through fluorescence enhancement, and the detection limit was 0.17 μM. The Zn-HTBA also selectively detect CrO42-/Cr2O72- and Fe3+ with the detection limits of 0.014/0.022 and 0.12 μM through fluorescence quenching. In addition, the Zn-HTBA was found to sensitively detect nitroaromatic compounds in ethanol through fluorescence quenching. The possible detection mechanisms were studied in detail through UV–Vis, PXRD, XPS, etc. The mechanism of Zn-HTBA to detect CrO42-/Cr2O72-, Fe3+ and 2-nitrotoluene was energy competition, and the mechanism of Zn-HTBA to detect lysine was the formation of hydrogen bonds.

Polymer organic framework-based ratiometric fluorescent probe for non-enzymatic glucose detection

Fluorescent pyrene-based polymer organic frameworks (POFs) PY-POFs were designed and synthesized by a facile solvothermal route for glucose detection, employing melamine, aromatic dialdehydes, 1-pyrenecarboxaldehyde, and phenylboronic acid as building blocks. The structure and photophysical properties of as-prepared POFs were systematically characterized via Fourier-transform infrared spectroscopy, X-ray photoelectron spectroscopy, scanning electron microscopy, transmission electron microscopy, N2 adsorption isotherms and fluorescence spectroscopy. The PY-POFs possessed intrinsic microporosity derived from extensively interlinked aminal structures, boasting specific surface areas reaching 192 m²/g, pore volumes extending to 0.8079 cm³/g, pore diameters of up to 17 nm, and an average particle dimension of ∼ 31 nm. In addition, the PY-POFs exhibited a dual-wavelength emission of pyrene excimer (λem = 472 nm) and monomer (λem = 382, 400, and 422 nm). When exposed to glucose, the former emission decreased while the latter increased, corresponding to the conformational transition of pyrene molecule from excimer to monomer. The non-enzymatic ratiometric fluorescent sensor displayed remarkable sensitivity towards glucose within 10−200 μM range, with a limit of detection (LOD) of 5.6 µM. Moreover, the glucose detection ability of PY-POFs was also in vivo demonstrated within zebrafish. Application of this sensor to real samples such as human saliva and DMEM biological matrix presented satisfactory results and good reproducibility. These results indicate that the fluorescent glucose sensor PY-POFs have broader prospects for utilization within biological studies.

Ultrasensitive Dual Fluorophore-Conjugated Carbon Dots for Intracellular pH Sensing in 3D Tumor Models.

The dysregulation of pH has been linked to the onset of chronic conditions, such as cancer and neurological diseases. Consequently, the development of a highly sensitive tool for intracellular pH sensing is imperative to investigate the interplay between pH and the biochemical changes accompanying disease pathogenesis. Here, we present the development of a ratiometric fluorescent nanoprobe, NpRhoDot, designed for precisely measuring pH levels. We demonstrate its efficacy in sensitively reporting intracellular pH in monolayer A549 lung cancer cells, primary fibroblast cells, and 3D tumor spheroids derived from the DLD-1 colorectal adenocarcinoma cell line. NpRhoDot leverages a novel design, where stable carbon dots are functionalized with a pH-responsive ratiometric fluorescent probe comprising a naphthalimide-rhodamine moiety, NpRho1. This design confers NpRhoDot with the high pH sensitivity characteristics of organic fluorescent probes, along with excellent photostability up to 1 h and biocompatibility of carbon dots. Through one-photon and two-photon fluorescence microscopy, we validate the reliability of NpRhoDot for biosensing intracellular pH in monolayer and three-dimensional tumor models from pH 4 to 7.

Imidazo[1,2-α]pyridine-based polarity and viscosity-dependent fluorescent probes and application in selective detection of 2,6-dichloro-4-nitroaniline

Three novel organic fluorescent probes M1-M3 with different conjugation length have been designed and synthesized from readily available starting materials via concise procedures. The obtained carbonyl-substituted imidazopyridines M1-M3 exhibit viscosity- and polarity-dependent dual-responsive properties. Structural modulation indicates that the polarity and viscosity sensitivity of M1-M3 can be tuned by variation of conjugation length. Importantly, M1-M3 demonstrate sensing ability for 2,6-dichloro-4-nitroaniline (DCN) with high selectivity and sensitivity in THF with a limit of detection (LOD) of 189 ppb, 62 ppb, and 54 ppb, respectively. The luminescence quenching of the compounds is caused by photo-induced electron transfer.

Application of novel AIE materials in fluorescence probes

Fluorescence bioimaging is considered an indispensable technique in biological research due to its high sensitivity, excellent spatial and temporal resolution, non-invasiveness, rapid and real-time responsiveness, as well as its ease of accessibility. Among these, fluorescence probes play a pivotal role in fluorescence imaging technology thanks to their strong specificity, high sensitivity, rapid response, and straightforward implementation. Aggregation-Induced Emission (AIE) molecules exhibit a unique phenomenon: they emit weak or no fluorescence when dissolved in a solution but become highly luminescent under aggregated/clustering conditions. Leveraging this, fluorescent probes with AIE characteristics often demonstrate fluorescence activation under conditions of spontaneous aggregation or binding with analytes, showcasing high sensitivity and exceptional signal-to-noise ratios. Consequently, fluorescence probes based on the distinctive optical properties of AIE hold immense potential for applications in fluorescence bioimaging. This paper primarily investigates the synthesis properties and applications in biosensing of a series of novel organic fluorescence probes with aggregation-induced emission (AIE) characteristics. By synthesizing a range of compounds with AIE properties and experimentally determining their photophysical properties and molecular structures, the study explores the potential applications of these compounds in DNA recognition, protein detection, small molecule recognition, cellular pH detection, and bioimaging.

A Fluorescent Organic Probe for Naked-Eye Detection of Chloromethanes.

An organic fluorescent probe (OFP-TAR) with a propeller-like structure was designed and synthesized. The photoluminescence of OFP-TAR in solution exhibited a significant red shift with the increase of solvent polarity, enabling a transition of fluorescence emission from blue (445 nm) to yellow (540 nm). The organic thin-film materials based on OFP-TAR/PMMA exhibit significant color changes upon exposure to CH2Cl2, CHCl3, and CCl4, with their maximum fluorescence wavelengths measured at 445, 471, and 494 nm respectively. The device facilitates the visual detection of chloromethanes and is capable of enduring more than 7 cycles of testing. These materials can also be prepared as binary-coded microarray data storage devices or applied in the field of anti-counterfeiting. The quantum yields of guest-loaded crystals CH2Cl2@OFP-TAR, CHCl3@OFP-TAR and CCl4@OFP-TAR are observed as 19.13 %, 8.79 %, and 0.83 % respectively, which are consistent with the tendency of OFP-TAR in CH2Cl2 (47.30 %), CHCl3 (34.27 %) and CCl4 (3.10 %). The fluorescence properties of OFP-TAR, OFP-TAR/PMMA, guest-loaded and guest-free crystals provided insights into the special response mechanism of OFP-TAR towards different chloromethanes.

An Eu (III)-functionalized covalent organic framework fluorescent probe for specific detection of Flumequine based on pore restriction and antenna effect

The overuse of quinolone antibiotics has led to a series of health and environmental issues. Herein, we combine the distinct luminescence properties of Eu3+ with the unique structure of covalent organic frameworks (COFs) to develop a precise and sensitive fluorescent probe for detecting Flumequine (Flu) in water. Eu3+ is thoroughly anchored into the channels of COFs as recognition sites, while the synthesized probe material still maintains its intact framework structure. The unique structure of COFs provides excellent support and protection for Eu3+. Therefore, COF-Eu can rapidly bind with Flu which can transfer the absorbed energy to Eu3+ through an “antenna effect”, resulting in red fluorescence. Moreover, there is a good linear relationship between Flu concentration in the range of 0–30 µM, with a detection limit of 41 nM. Simultaneously, the material maintains remarkable reproducibility, with its performance remaining almost unchanged after five cycles of use. Remarkably, the probe demonstrates excellent Flu recovery rates in real samples. This study provides a viable approach for the recognition of flumequine in the environment through a customized fluorescence detection method.

Novel fluorescence probe for ClO− in living cells: Based on FRET mechanism

Hypochlorous acid (HClO) as a kind of reactive oxygen species (ROS) plays a vital role in many biological processes. Organic fluorescence probes have attracted great interests for the detection of HClO, due to their relatively high selectivity and sensitivity, satisfactory spatiotemporal resolution and good biocompatibility. Constructing fluorescence probes to detect HClO with advantages of large Stokes shift, wide emission gap, near infrared emission and good water solubility is still challenging. In this work, a new ratiometric fluorescence probe (named HCY) for HClO was developed. FRET-based HCY was constructed by bonding a coumarin and a flavone fluorophore. In absence of HClO, HCY exists FRET process, however, FRET is inhibited in the presence of HClO because the conjugated double bond broke. Due to the good match of the emission spectrum of the donor and the absorption spectrum of the acceptor, the FRET system appears favorable energy transfer efficiency. HCY showed high sensitivity and rapid response time. The linearity between the ratios of fluorescence intensity and concentration of HClO was established with a low limit of detection. What’s more, HCY was also applied for fluorescence images of HClO in RAW264.7 cells.

Recent Advances in Organic Small-Molecule Fluorescent Probes for the Detection of Zinc Ions (Zn2+).

Zinc(II) ions (Zn2g) play crucial roles in the growth, propagation, and metabolism of animals, plants, and humans. Abnormal concentrations of Zn2+ in the environment and living organisms pose potential risks to environmental protection and human health. Therefore, it is imperative to develop rapid, reliable and in-situ detection methods for Zn2+in both environmental and biological contexts. Furthermore, effective analytical methods are required for diagnosing diseases and understanding physiological metabolic mechanisms associated with Zn2+concentration levels. Organic small-molecule fluorescent probes offer advantages such as fast, reliable, convenient, non-destructive detection capabilities and have significant application potential in Zn2+ detection and bioimaging; thus garnering extensive attention. Over the past two years alone, various organic small-molecule probes for Zn2+ based on different detection mechanisms and fluorophores have been rapidly developed. However, these probes still exhibit several limitations that need further resolution. In light of this context, we provide a comprehensive summary of the detection mechanisms, performance characteristics, and application scope of Zn2+ fluorescence probes since year 2022 while highlighting their advantages. We also propose solutions to address existing issues with these probes and outline future directions for their advancement. This review aims to serve as a valuable reference source offering insights into the development of advanced organic small-molecule-based fluorescence probes specifically designed for detecting Zn2+.

Solvent effects on the ESIPT emission of salicylaldehyde Schiff base derivative: A theoretical reconsideration

Salicylaldehyde Schiff base derivatives have potential applications in bioimaging, chemical sensors and organic luminescence materials because of their high fluorescence quantum yield with the excited-state intramolecular proton transfer (ESIPT) process. Recently, Shu et al. studied the luminescent mechanism of the Et2N-substituted salicylaldehyde Schiff base compound (DDHAC) in solvents [Dyes and Pigments 195 (2021) 109708]. It showed double-peaked bands in an n-hexane solvent but a single-peaked band in an acetonitrile solvent. They suggested that the open-enol in the acetonitrile solvent was the dominant species. They considered that open-enol fluorescence and ESIPT fluorescence constituted a dual fluorescent phenomenon in the n-hexane solvent. However, because acetonitrile is a non-protonic solvent, DDHAC molecules in acetonitrile mainly existed as the lower-energy hydrogen-bonded enol form rather than the open-enol form alone. In addition, the open-enol did not undergo the ESIPT process in n-hexane. Therefore, the luminescence mechanism of the DDHAC molecules in n-hexane and acetonitrile solvents needs to be reconsidered. In this study, we investigated the DDHAC molecules in n-hexane and acetonitrile solvents using the density functional theory and time-dependent density functional theory. The potential energy surfaces and optimisation of structures elucidated that the DDHAC molecules in n-hexane and acetonitrile solvents underwent the ESIPT process. The calculated fluorescence peaks demonstrated that the single-peaked broad emission band in the acetonitrile solvent was formed by the combination of the hydrogen-bonded enol* and keto* forms rather than open-enol*. Moreover, the dual fluorescence peaks in the n-hexane solvent were reattributed to the open-enol* form and hydrogen-bonded enol* form. The lack of keto* fluorescence in the n-hexane solvent was attributed to diminished charge coupling in comparison to the enol* form. Our results revise the mechanism of the DDHAC molecules in n-hexane and acetonitrile solvents, providing guidance for designing efficient organic fluorescence probes.

Activatable NIR Fluorescence Probe for Epinephrine Detection and Bioimaging Based on Anionic Heptamethine Cyanine.

Epinephrine (EP) is an essential catecholamine in the human body. Currently, most EP detection methods are not suitable for in vivo detection due to material limitations. An organic small molecule fluorescent probe based on a chemical cascade reaction for the detection of EP was designed. Anionic heptamethine cyanine dye was selected as a fluorescent dye because of its NIR fluorescence emission with excellent biocompatibility. The secondary amine of EP nucleophilically attacks the carbonate of the probe with its stronger nucleophilicity and further undergoes intramolecular nucleophilic cyclization to release the fluorophore. Other substances containing only primary amines or no β-OH lack reaction competitiveness due to their weaker nucleophilicity or inability to undergo further cyclization. The fluorescence recovery of the probe was linearly related to the EP concentration of 2-75 μmol/L. The detection limit was 0.4 μmol/L. The recovery rate was 94.78-111.32%. Finally, we successfully achieved bioimaging of EP in living cells and EP analogue in nematodes.

Advances in Organic Fluorescent Probes for Intracellular Zn2+ Detection and Bioimaging.

Zinc ions (Zn2+) play a key role in maintaining and regulating protein structures and functions. To better understand the intracellular Zn2+ homeostasis and signaling role, various fluorescent sensors have been developed that allow the monitoring of Zn2+ concentrations and bioimaging in live cells in real time. This review highlights the recent development of organic fluorescent probes for the detection and imaging of intracellular Zn2+, including the design and construction of the probes, fluorescent response mechanisms, and their applications to intracellular Zn2+ detection and imaging on-site. Finally, the current challenges and prospects are discussed.

On-Site Quantitative Visualization of Singlet Oxygen in Crops via an Organic Small Molecule-Based Ratiometric Fluorescent Probe and a Mobile Fluorescence Analysis Device.

Singlet oxygen (1O2) plays imperative roles in a variety of biotic or abiotic stresses in crops. The change of its concentration within a crop is closely related to the crop growth and development. Accordingly, there is an urgent need to develop an efficient analytical method for on-site quantitative detection of 1O2 in crops. Here, we judiciously constructed a novel ratiometric fluorescent probe, SX-2, for the detection of 1O2 in crops. Upon treating with 1O2, probe SX-2 displayed highly selective ratiometric fluorescence response, which is favorable for the quantitative detection of 1O2. Concurrently, the fluorescence solution color of probe SX-2 was varied, obviously from blue to yellow, indicating that the probe is beneficial for on-site detection by the naked eye. Sensing reaction mechanism studies showed that the 2,3-diphenyl imidazole group in SX-2 could function as a new selective recognition group for 1O2. Probe SX-2 was utilized for the detection of photoirradiation-induced 1O2 and endogenous 1O2 in living cells. The changes in the 1O2 level in zebrafish were also tracked by fluorescence imaging. In addition, the production of 1O2 in crop leaves under a light source of different wavelengths was studied. The results demonstrated more 1O2 were produced under a light source of 365 nm. Furthermore, to achieve on-site quantitative detection, a mobile fluorescence analysis device has been made. Probe SX-2 and mobile fluorescence analysis device were capable of on-site quantitative detecting of 1O2 in crops. The method developed herein will be convenient for the on-site quantitative measurement of 1O2 in distinct crops.

Theoretical Insights into the Effect of Different Numbers of Thiophene Groups on Hydrogen Bond Interaction and Excited-State Intramolecular Proton-Transfer Process for Flavonoid Derivatives.

In this study, we systematically explored the impact of varying the number of thiophene groups on the hydrogen bond interaction and excited-state intramolecular proton-transfer (ESIPT) processes in flavonoid derivatives (STF, DTF, and TTF) using the density functional theory and time-dependent density functional theory methods. Initially, a thorough analysis of the optimized geometric structures revealed that the intramolecular hydrogen bond in the S1 state is enhanced and gradually weakened as the number of thiophene groups increases. To gain a deeper understanding of the hydrogen bond interaction, topological analysis, interaction region indicator scatter plots, and isosurface plots were employed. These images provide further insights that align with the structural analysis. Additionally, we observed a red-shift in the electronic spectra (absorption and fluorescence spectra), which is primarily attributed to the narrowing of the energy gap between the highest occupied molecular orbital and the lowest unoccupied molecular orbital, as elucidated by the frontier molecular orbitals. Furthermore, a combined analysis between the hole-electron distribution and the transition density matrix heat map shows that electron excitation involves the unidirectional charge-transfer mechanism. In the end, by conducting relaxed potential energy curve scans, we found that an increase in the number of thiophene groups elevates the energy barrier for ESIPT, making it more challenging for the reaction. In summary, our study underscores the vital effect of thiophene-substituted numbers in modulating the ESIPT process, which is able to provide valuable insights for the design and synthesis of desired organic fluorescent probes in the future.

A high-performance organic fluorescent probe with aggregation-induced emission properties for long-term tumor monitoring

Near-infrared organic fluorescent probes have great need in biological sciences and medicine but most of them are still largely unable to meet demand. In this work, a delicate multipurpose organic fluorescent probe (DPPM-TPA) with aggregation-induced emission performances is designed and prepared by facile method to reflect fluorescence labeling, two-photon imaging, and long-term fluorescent tracking. Specifically, DPPM-TPA NPs was constructed from 4-(diphenylamino)phenylboronic acid and DPPM-Br by classical Suzuki coupling reaction and then coated with F127. Such nanoprobe possessed high stability in diverse medium under ambient temperatures, low cytotoxicity, and brilliant fluorescence performance. More importantly, DPPM-TPA NPs showed excellent two-photon imaging and extraordinary long-term fluorescence tracing capacity to malignant tumor, and it can last up to 9 days. These results indicated that DPPM-TPA NPs is expected to serve as a fluorescent probe for photodiagnostic and providing a new idea for the development of long-term fluorescent tracker.

Synthesis of nanoflower-shaped covalent organic framework fluorescent probe for sensitive detection of aluminum ions

The widespread application of aluminum has led to numerous adverse effects on organisms due to aluminum pollution. In this study, we synthesized a two-dimensional fluorescent covalent organic framework (TAPB-DMTP-COF) material with nanoflower morphology for fluorescence detection of aluminum ions. The coordination binding of Al3+ with the nitrogen atoms on the imine bonds and the oxygen atoms on the methoxy groups restricts intramolecular rotation in TAPB-DMTP-COF, which inhibits the photo-induced electron transfer (PET) in the COF molecule and triggers the fluorescence emission "turn on" phenomena. A fluorescence method for Al3+ detection was established, and it was found that the method exhibits high sensitivity, selectivity, a wide detection range from 4.0 to 100.0 μM, and a low limit of detection (LOD) of 38.5 nM. This work provided new ideas for the convenient and rapid detection of aluminum ions in environmental analysis and expanded the application of fluorescence COFs in analytical sensing.

A Halloysite Nanotubes-based Probe for Efficient Fluorescence Detection and Adsorption Removal of Pb2+ in Water.

The detection and removal of Pb2+ is of utmost importance for environmental protection and human health due to its toxicity, persistent pollution, and bioaccumulation effects. To address the limitations associated with organic small molecule-based fluorescence probes such as poor water solubility and single functionality in detecting Pb2+, a fluorescence probe based on halloysite nanotubes was developed. This probe not only enables specific, rapid, and reliable detection of Pb2+ but also facilitates efficient removal of it from water. The development of this bifunctional fluorescent probe provides a valuable insight for designing more advanced probes targeting heavy metal ions.

Excited-State Intramolecular Proton Transfer-Driven Photon-Gating for Photoelectrochemical Sensing of CO-Releasing Molecule-3.

Different from prevalent approaches such as immunological recognition, complementary base pairing, or enzymatic regulation in current photoelectrochemical (PEC) sensing, this study reported an excited-state intramolecular proton transfer (ESIPT)-driven photon-gating PEC sensor. The sensor is developed for the detection of CO-releasing molecule-3 (CORM-3) by modifying an ESIPT-switched organic fluorescent probe molecule (NDAA) onto the surface of a p-type semiconductor (BiOI). The NDAA can be excited and exhibit strong green fluorescence after responding with CORM-3, resulting in an electrode-interface photon competitive absorption effect due to the switch on ESIPT and considerably reducing the photocurrent signal. The experimental results revealed that the as-developed PEC sensor achieved good analytical performance with high selectivity and sensitivity, with a linear range of 0.01-1000 μM and a lower detection limit of 6.5 nM. This work demonstrates the great potential of the organic fluorescent probe molecule family in advancing PEC analysis. It is anticipated that our findings will stimulate the creation of diverse functional probes possessing distinctive characteristics for inventive PEC sensors.

Tumor Microenvironment Activatable Nanoprodrug System for In Situ Fluorescence Imaging and Therapy of Liver Cancer.

The development of new imaging and treatment nanoprodrug systems is highly demanded for diagnosis and therapy of liver cancer, a severe disease characterized by a high recurrence rate. Currently, available small molecule drugs are not possible for cancer diagnosis because of the fast diffusion of imaging agents and low efficacy in treatment due to poor water solubility and significant toxic side effects. In this study, we report the development of a tumor microenvironment activatable nanoprodrug system for the diagnosis and treatment of liver cancer. This nanoprodrug system can accumulate in the tumor site and be selectively activated by an excess of hydrogen peroxide (H2O2) in the tumor microenvironment, releasing near-infrared solid-state organic fluorescent probe (HPQCY-1) and phenylboronic acid-modified camptothecin (CPT) prodrug. Both HPQCY-1 and CPT prodrugs can be further activated in tumor sites for achieving more precise in situ near-infrared (NIR) fluorescence imaging and treatment while reducing the toxic effects of drugs on normal tissues. Additionally, the incorporation of hydrophilic multivalent chitosan as a carrier effectively improved the water solubility of the system. This research thus provides a practical new approach for the diagnosis and treatment of liver cancer.