Multifunctional Probe Research Articles

A multifunctional near-infrared fluorescent probe based on benzothiazole structure for fluoride-ion detection

Fluoride ions (F−) are one of the essential trace elements for the human body and are widely existed in nature. In this study, we present a novel fluorescent probe (YF-SZ-F) designed and synthesized for the specific detection of F−. The probe exhibits high sensitivity, excellent selectivity, and low cytotoxicity, making it a promising tool for biomedical applications. Imaging experiments conducted on zebrafish and Arabidopsis roots demonstrate the probe’s remarkable cellular permeability and biocompatibility, laying a solid foundation for its potential biomedical utility. Furthermore, the probe holds potential for practical applications in environmental monitoring and public health through its capability to detect fluoride ions in water samples and via mobile phone software. This multifaceted functionality underscores the broad applicability and significance of the fluorescent probe, not only in scientific research but also in real-world scenarios, contributing to the development of more convenient and precise methods for fluoride detection.

SERS surgical navigation with postsurgical immunotherapy of local microtumors and distant metastases for improved anticancer outcomes.

The standard of clinical care of most malignant solid cancers is surgery, followed by postsurgical adjuvant therapy, but microtumor lesions left behind after surgery and invisible distant metastases are the major reasons for treatment failure. Here, we report an integrated strategy combining surface-enhanced Raman spectroscopy (SERS) surgical navigation with postsurgical immunotherapy elicited by near-infrared II photothermal treatment and programmed death-1 antibody. The SERS surgical navigation is principally based on the multifunctional optical probes (namely, MATRA probes) integrating with T1-weighted magnetic resonance (MR) imaging, photothermal effect and Raman spectroscopic detection. We demonstrate in a 4T1 breast tumor mouse model that the pre-surgical MR/SERS dual-modal imaging is capable of providing comprehensive tumor information, and intraoperative SERS detection allows accurately delineating the tumor margins and guiding the surgical resection in real time with the least residual microscopic foci. We verify that the postsurgical immunotherapy effectively eradicates those local microtumor lesions and invisible distant metastases, greatly inhibiting the postsurgical cancer recurrence and distant metastasis.

Development of a monomolecular multifunctional imaging probe for dual labeling of LDL to image and characterize atherosclerotic plaques

Development of a monomolecular multifunctional imaging probe for dual labeling of LDL to image and characterize atherosclerotic plaques

Bioimaging research of a turn-on homodimer fluorescent probes for pH and viscosity: A new turn-on strategy

A turn-on homodimer fluorescent probe (BL) that responded to pH and viscosity was facilely synthesized based on homo-FRET. The probe exhibited strong fluorescence intensity and color changes in alkaline pH. In addition, the probe's fluorescence had altered with changes in viscosity. What's more, Mitochondrial localization and pH bioimaging in E. coli cell experiments confirmed that BL was a novel multifunctional probe that could be applied to mitochondrial-targeted fluorescence imaging under normal conditions and real-time fluorescence imaging of E. coli under alkaline conditions.

Dual excitation channel ratiometric fluorescent probes for visual and fluorescent detection of anthrax spore biomarker and tetracycline hydrochloride

Long-term and excessive use of tetracycline hydrochloride (TC) can lead to its accumulation in the environment, which can cause water contamination, bacterial resistance, and food safety problems. 2,6-Pyridine dicarboxylic acid (DPA) is a major biomarker of Bacillus anthracis spores, and its rapid and sensitive detection is of great significance for disease prevention and counter-terrorism. A bifunctional ratiometric fluorescent nanoprobe has been fabricated to detect DPA and TC. 3,5-dicarboxyphenylboronic acid (BOP) was intercalated into layered europium hydroxide (LEuH) by the ion-exchange method and exfoliated into nanosheets as a fluorescent nanoprobe (PNP). DPA and TC could significantly enhance the red fluorescence of Eu3+ through the antenna effect under different excitation wavelengths, while the fluorescence of BOP can be used as a reference based on the constant emission intensity, realizing ratiometric detection. A low limit of detection (LOD) for the target (DPA: 9.7 nM, TC: 21.9 nM) can be achieved. In addition, visual detection of DPA and TC was realized using color recognition software based on the obvious color changes. This is the first ratiometric fluorescent nanoprobe based on layered rare-earth hydroxide (LRH) for the detection of DPA and TC simultaneously, which opens new ideas in the design of multifunctional probes.

Novel benzothiazole-based fluorescent probe for efficient detection of Cu2+/S2− and Zn2+ and its applicability in cell imaging

BackgroundIn recent years, environmental pollution has been increasing due to the excessive emission of toxic ions, which has caused serious harm to human health and ecological environment. There are various methods for detecting Cu2+, S2− and Zn2+, but the traditional ion detection methods have obvious disadvantages, such as poor selectivity and long detection time. Therefore, it is still crucial to develop simple, efficient and rapid detection methods. ResultsA fluorescent probe based on benzothiazole, (E)-N'-(3-(benzo[d]thiazol-2-yl)-2-hydroxy-5-methylbenzylidene)-3,4,5-tris(benzyloxy)benzohydrazide (BT), was designed and synthesized. It was characterized using ESI-MS, 1H NMR, and 13C NMR. BT can be used as a chemosensor to detect Cu2+, S2− and Zn2+ in CH3CN/H2O (7:3, v/v, pH = 7.4, HEPES buffer: 0.1 M), with detection limits of 0.301 μM, 0.017 μM, and 0.535 μM, respectively. At an excitation wavelength of 320 nm, BT exhibits an “on-off-on” response to Cu2+/S2− and enhanced fluorescence response to Zn2+, with a change in fluorescence color from orange to green. The coordination ratio of ions to the probe was determined to be 1:1 through Job's plot and hydrogen spectral titration. The recognition mechanism was discussed in conjunction with theoretical calculations. Furthermore, the probe has been successfully used in test strips and medical swabs colorimetry, as well as live cell imaging. SignificanceThe probe BT lays the foundation for the design and synthesis of multifunctional fluorescent probes. As a portable detection method, probe BT was used to detect Cu2+, S2− and Zn2+ on strips. Furthermore, the probe was applied to biological cells to detect target ions with low cytotoxicity and excellent cell permeability. This indicating that it can be used as a potential candidate for tracking Cu2+ and S2− in clinical diagnostics and biological systems.

An advanced 3D DNA nanoplatform for spatiotemporally confined enhanced dual-mode biosensing MicroRNA in cancer cell

Dual-mode signal output platforms have demonstrated considerable promise due to their improved anti-interference capability and inherent signal self-correction. Nevertheless, traditional discrete-distributed signal probes often encounter significant drawbacks, including limited mass transfer efficiency, diminished signal strength, and instability in intricate biochemical environments. In response to these challenges, a scalable and hyper-compacted 3D DNA nanoplatform resembling “periodic focusing heliostat” has been developed for synergistically enhanced fluorescence (FL) and surface-enhanced Raman spectroscopy (SERS) biosensing of miRNA in cancer cells. Our approach utilized a distinctive assembly strategy integrating gold nanostars (GNS) as fundamental “heliostat units” linked by palindromic DNA sequences to facilitate each other hand-in-hand cascade alignment and condensed into large scale nanostructures. This configuration was further augmented by the incorporation of gold nanoparticles (GNP) via strong Au–S bonds, resulting in a sturdy framework for improved signal transduction. The initiation of this assembly process was mediated by the hybridization of dsDNA to miRNA-21, which served as a primer for polymerization and nicking reactions, thus generating a multifunctional T2 probe. This probe is intricately designed with three distinct parts: a 3′-palindromic end for structural integrity, a central region for capturing SERS-active probes (Cy3-P2), and a 5′-segment for attaching fluorescence reporters. Upon integration T2 into the GNS-based heliostat unit, it promotes palindromic arm-induced aggregation and plasma exciton coupling between plasma nanoparticles and signal transduction tags. This clustered arrangement creates a high-density “hot spot” array that maximizes the local electromagnetic fields necessary for enhanced SERS and FL response. This superstructure supports enhanced aggregation-induced signal amplification for both SERS and FL, offering exceptional sensitivity with LOD as low as 0.0306 pM and 0.409 pM. The efficacy of this method was demonstrated in the evaluation of miRNA-21 in various cancer cell lines.

Near-infrared and multifunctional fluorescent probe enabled by cyanopyridine cyanine dye for bisulfite recognition and biological imaging

SO2 derivatives, sulfite/bisulfite, are widely employed in both the food processing and drug synthesis industries. Despite their widespread application, excessive levels of sulfite/bisulfite can negatively impact human health. Most probes for detecting sulfite/bisulfite are restricted by their fluorescence within the visible spectrum range and poor solubility in aqueous solution, which limit their use in food testing and biological imaging. Herein, a near-infrared probe comprising of the cyanopyridine cyanine skeleton, 4-((Z)−2-((E)−2-chloro-3-(2-cyano-2-(1-methylpyridine-4(1H)-ylidene)ethylidene)cyclohex-1-en-1-yl)−1-cyanovinyl)−1-methylpyridin-1-ium (abbreviated as CCP), was developed. This probe enables precise quantification of bisulfite (HSO3-) in almost pure buffered solutions, showing a near-infrared fluorescence emission at 784 nm with an impressively low detection limit of 0.32 μM. The probe stands out for its exceptional selectivity, minimal susceptibility to interference, and strong adaptability. The probe CCP utilizes the CC bond to trigger a near-infrared fluorescence quenching reaction with HSO3- via nucleophilic addition, which effectively disrupts the large delocalization within the molecule for accurate HSO3- identification. Moreover, the probe has been successfully applied in detecting HSO3- in various food products and living cells, simplifying the measurement of HSO3- content in water samples. This advancement not only enhances the analytical capabilities but also contributes to ensuring food safety and environmental protection. Environmental ImplicationSO2 derivatives including sulfite/bisulfite, serving dual roles as preservatives and antioxidants, have widespread application across various sectors including food preservation, water sanitation, and the pharmaceutical industry. Despite their widespread application, excessive levels of sulfite/bisulfite can affect human health. Developing methods for precisely and sensitively detecting sulfite/bisulfite in food products and biological samples is important for ensuring food safety and environmental protection. Here, a sensitive near-infrared and multifunctional fluorescent probe in a 99.9 % buffered solution, along with water gel encapsulation, has been successfully applied for the detection of bisulfite in food, authentic water samples, and biological cells.

A europium-based CP fluorescent probe for sensing malachite green, ascorbic acid and uric acid

Lanthanide (such as Eu and Tb) coordination polymers (CPs) having various coordination modes and unique fluorescence properties can be used as highly sensitive chemical sensors. We synthesized a europium-based CP of Eu2(PEDA)3(H2O)4 (referred to as Eu-PEDA, PEDA = 1,2-phenylenediacetic acid group), and investigated its application in fluorescence detection. Eu-PEDA can be considered as a multifunctional fluorescent probe for sensing malachite green (MG), ascorbic acid (AA) and uric acid (UA) to achieve rapid fluorescence quenching response, and Eu-PEDA exhibited high quenching efficiency Ksv (6.21 × 105 M−1 for MG; 1.16 × 104 M−1 for AA; 1.02 × 104 M−1 for UA), low detection limit (MG: 0.050 μM; AA: 0.705 μM; UA: 0.965 μM). Furthermore, Eu-PEDA suspension dipping test strips, the Eu-PEDA test strips are obtained. The resultant test strip, shows the potential for directly visual detection of MG, AA and UA just by dripping the analytes on it. This work thus offers a facile and efficient strategy for rapid and visual detection of MG, AA and UA by Eu-based coordination compounds. In addition, the possible mechanisms of fluorescence quenching were studied by absorption spectra, the emission and excitation spectra.

A Water-Soluble Multifunctional Probe for Colorimetric Copper Sensing, Lysosome Labelling and Live-Cell Imaging.

We report a water-soluble fluorescence and colorimetric copper probe (LysoBC1); this system can also serve for lysosome labeling and for the dynamic tracking of Cu2+ in living cells. The sensing mechanism takes advantage of the synergic action by the following three components: i) a lysosome targeting unit, ii) the spirolactam ring-opening for the selective copper chelation and iii) the metal-mediated hydrolysis of the rhodamine moiety for fluorescence enhancement. In aqueous environment the molecule acts as a fluorescent reversible pH sensor and as colorimetric probe for Cu2+ at physiological pH; the hydrolysis of the copper targeting unit resulted in a 50-fold increase of the fluorescence intensity. Most importantly, in vitro cell analyses in undifferentiated (SH SY5Y) and differentiated (d-SH SY5Y) neuroblastoma cells, LysoBC1 is able to selectively accumulate into lysosome while the copper binding ability allowed us to monitor intracellular copper accumulation into lysosome.

Multifunctional nanoenzyme lateral flow immunoassay strip for rapid and ultrasensitive detection of carbofuran in vegetables

A lateral flow immunoassay strip (LFIAS) is one of the most frequently rapid test technologies for carbofuran (CAR). Nevertheless, the LFIAS has a poor quantitative capability and low sensitivity. And, it also requires often complex sample handling steps, making testing time longer. In this study, Fe3O4 nanoparticles were successively modified with MIL-100(Fe)-based metal-organic framework (MOF) and chloroplatinic acid hexahydrate to obtain a core-shell complex of Fe3O4-MOF-Pt. The complex had a peroxidase-mimicking activity catalytic function that enabled signal amplification and sensitivity enhancement. Upon coupling with carbofuran monoclonal antibody (CAR-mAb), the magnetic separation properties of the probe enabled target-specific enrichment. The LFIAS based on Fe3O4-MOF-Pt nanocomposites could detect CAR in the range of 0.25–50 ng mL−1 with a limit of detection (LOD) of 0.15 ng mL−1, enabling colorimetric and catalytic analysis. In addition, the method showed high specificity and stability for detecting CAR in various vegetables, and recovery rates of the spiked samples were 91.40%−102.40%. In conclusion, this study provided one-stop detection of “target enrichment-visual inspection”. While lowering the LOD, it reduced the detection time and improved the detection efficiency. The multifunctional Fe3O4-MOF-Pt nanocomposite provides an idea for the construction of novel multifunctional probes to improve the detection performance of conventional LFIAS.

Methylene blue-encapsulated Cu(I)/(Ⅱ) mixed-valence MOF with spontaneously recycled catalysis for electrochemical-colorimetric dual-signal readout aptasensor

The sensitive and accurate detection of thrombin (Tb) is of significance for early diagnosis of thrombosis, vascular injuries, and cancers. A dual-signal aptasensor with effective signal amplification is a promising approach to improve accuracy and sensitivity simultaneously for detection of Tb. Herein, an electrochemistry (EC)-colorimetry dual-readout aptasensor was proposed for Tb detection based on a multifunctional probe, MB@ZIF-90/Cu(I)/(Ⅱ). Methylene blue (MB), as an electroactive and colorimetric dual-signal molecule, was encapsulated into ZIF-90 along with introducing Cu species to form MB@ZIF-90/Cu. After post-treatment heating, MB@ZIF-90/Cu(I)/(Ⅱ) formed with the catalysis toward MB even without the addition of substrate. The catalysis was further enhanced by the “confinement effect” of Cu(I)/Cu(Ⅱ) system and MB in the pore for amplified electrochemical signals. While, the unbounded probes were collected for colorimetric analysis. As a result, EC-colorimetric dual-readout aptasensor exhibits wider linear ranges of 1 pM–10 μM and 0.1 nM–100 μM and lower detection limit down to 0.3 pM and 0.03 nM for Tb, respectively. The mixed-valence MOFs-based dual-signal aptasensor achieved ultrasensitive detection of Tb with more reliable results, revealed its promising application in clinical diagnosis of Tb and even other biomarkers.

A dual-response fluorescent probe to reveal the role of ferroptosis in drug-induced liver injury

Drug-induced liver injury (DILI) has already been recognized as a formidable obstacle in global health issues. Despite the proposed common cellular death types in DILI, exploration of alternative mechanisms such as ferroptosis for prevention and therapy remains limited. Visualization of the role of ferroptosis in DILI holds profound promise in advancing diagnostic and therapeutic strategies for DILI. To address this, herein, a unique multi-functional fluorescent probe, VIS-HNO, was elaborately built up and endowed with the extraordinary performance of excellent selectivity and high sensitivity toward HNO and viscosity. Employing the probe VIS-HNO with the confocal fluorescence imaging, the up-regulations of HNO and viscosity in the process of DILI and ferroptosis were disclosed. Then the remediation of DILI was achieved by VIS-HNO under the treatment of ferrostatin1 (Fer-1), indicating its therapeutic value for DILI. In addition, taking into account the significance of HNO in determining the role of ferroptosis in DILI, the pathway of HNO production in DILI was further explored, which presented essential insights into the role of ferroptosis in DILI. Overall, these findings demonstrated that VIS-HNO may be a potent tool to aid in diagnosing ferroptosis and DILI, more importantly, to understand the role of ferroptosis in DILI by evaluating HNO and viscosity level, which will facilitate the exploration of unique strategies for DILI treatment.

Phenylboronic Acid-Modified Membrane-Like Magnetic Quantum Dots Enable the Ultrasensitive and Broad-Spectrum Detection of Viruses by Lateral Flow Immunoassay.

Although lateral flow immunochromatographic assay (LFIA) is an effective point-of-care testing technology, it still cannot achieve broad-spectrum and ultrasensitive detection of viruses. Herein, we propose a multiplex LFIA platform using a two-dimensional graphene oxide (GO)-based magnetic fluorescent nanofilm (GF@DQD) as a multifunctional probe and 4-aminophenylboronic acid (APBA) as a broad-spectrum recognition molecule for viral glycoprotein detection. GF@DQD-APBA with enhanced magnetic/fluorescence properties and universal capture ability for multiple viruses was easily prepared through the electrostatic adsorption of one layer of density-controlled Fe3O4 nanoparticles (NPs) and thousands of small CdSe/ZnS-MPA quantum dots (QDs) on a monolayer GO sheet followed by chemical coupling with APBA on the QD surface. The GF@DQD-APBA probe enabled the universal capture and specific determination of different target viruses on the test strip through an arbitrary combination with the antibody-modified LFIA strip, thus greatly improving detection efficiency and reducing the cost and difficulty of multiplex LFIA for viruses. The proposed technique can simultaneously and sensitively diagnose three newly emerged viruses within 20 min with detection limits down to the pg/mL level. The excellent practicability of GF@DQD-APBA-LFIA was also demonstrated in the detection of 34 clinical specimens positive for SARS-CoV-2, revealing its potential for epidemic control and on-site viral detection.

Recent Progress of Multifunctional Molecular Probes for Triple-Negative Breast Cancer Theranostics.

Breast cancer (BC) poses a significant threat to women's health, with triple-negative breast cancer (TNBC) representing one of the most challenging and aggressive subtypes due to the lack of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) expression. Traditional TNBC treatments often encounter issues such as low drug efficiency, limited tumor enrichment, and substantial side effects. Therefore, it is crucial to explore novel diagnostic and treatment systems for TNBC. Multifunctional molecular probes (MMPs), which integrate target recognition as well as diagnostic and therapeutic functions, introduce advanced molecular tools for TNBC theranostics. Using an MMP system, molecular drugs can be precisely delivered to the tumor site through a targeted ligand. Real-time dynamic monitoring of drug release achieved using imaging technology allows for the evaluation of drug enrichment at the tumor site. This approach enables accurate drug release, thereby improving the therapeutic effect. Therefore, this review summarizes the recent advancements in MMPs for TNBC theranostics, encompassing the design and synthesis of MMPs as well as their applications in the field of TNBC theranostics.

Lipid droplets-targeting multifunctional fluorescent probe and its application in ferroptosis and bioimaging

Unraveling the inherent association between diseases and hypochlorous acid (HOCl), within the microenvironments of lipid droplets (LD) remains a formidable task due to the scarcity of research on multiple disease models. Intracellular HOCl and microenvironments (such as viscosity and polarity), are crucial in upholding redox balance, orchestrating diffusion, transportation and signal transduction within living cells. Nevertheless, the simultaneous visualization of HOCl alongside viscosity and polarity within the milieu of ferroptosis, inflammation, and neoplasia remains unachieved due to the absence of appropriate analytical tools. In response to this pressing need, we have developed an innovative, multifunctional near infrared (NIR) fluorescent probe, termed NCN, capable of simultaneously monitoring HOCl, viscosity, and polarity within lipid droplets. NCN is characterized by its heightened sensitivity to increased viscosity and decreased polarity, which allows it to target LDs with high specificity. Furthermore, the combined application of HOCl, viscosity, and polarity as diagnostic markers for cancer has been triumphantly demonstrated, encompassing cellular and tissue strata as well as in vivo tumor-bearing mouse models. Therefore, our probe offers a reliable means for understanding LD microenvironments and proposes a promising approach for the diagnostic evaluation of diseases.

Aversatile MOF as an electrochemical/fluorescence/colorimetric signal probe for the tri-modal detection of MMP-9 secretion in the extracellular matrix to identify the efficacy of chemotherapeutic drugs

BackgroundMMP-9 plays a crucial role in regulating the degradation of proteins within the extracellular matrix (ECM). This process closely correlates with the occurrence, development, invasion, and metastasis of various tumors, each exhibiting diverse levels of MMP-9 expression. However, the accuracy of detection results using the single-mode method is compromised due to the coexistence of multiple biologically active substances in the ECM. ResultsTherefore, in this study, a tri-modal detection system is proposed to obtain more accurate information by cross-verifying the results. Herein, we developed a tri-modal assay using the ZIF-8@Au NPs@S QDs composite as a multifunctional signal probe, decorated with DNA for the specific capture of MMP9. Notably, the probe demonstrated high conductivity, fluorescence response and mimicked enzyme catalytic activity. The capture segments of hybrid DNA specifically bind to MMP9 in the presence of MMP9, causing the signal probe to effortlessly detach the sensor interface onto the sample solution. Consequently, the sensor current performance is weakened, with the colorimetric and fluorescent signals becoming stronger with increasing MMP9 concentration. Notably, the detection range of the tri-modal sensor platform spans over 10 orders of magnitude, verifying notable observations of MMP-9 secretion in four tumor cell lines with chemotherapeutic drugs. Furthermore, the reliability of the detection results can be enhanced by employing pairwise comparative analysis. SignificanceThis paper presents an effective strategy for detecting MMP9, which can be utilized for both the assessment of MMP-9 in cell lines and for analyzing the activity and mechanisms involved in various tumors.

Fabrication of a novel 3D cadmium−organic framework doped with europium as multifunctional fluorescence probes for enhancing detectability

A novel 3D cadmium−organic framework (Cd-MOF) was prepared based on ionothermal synthesis method and successfully doped with Eu3+ cations to obtain Eu@Cd-MOF by postsynthetic modification method. Cd-MOF has a strong and stable framework due to the balance of cations on the overall structure. Using characterization methods such as powder X-ray powder diffraction (PXRD), Fourier transform infrared (FTIR) spectroscopy, and thermogravimetry analysis (TGA), these two materials have excellent solvent and chemical stability. More interestingly, both two materials can be used as multifunctional fluorescent probes to detect Fe3+, Co2+, Cr2O72−, MnO4−, and NB, and exhibit significant sensitivity, anti-interference, and recyclability. It is worth noting that compared to the original Cd-MOF, Eu@Cd-MOF enhanced excellent quenching effect on Fe3+, Co2+, Cr2O72−, MnO4−, and NB. Related to this, we conducted detailed research and discussion on possible fluorescence sensing mechanisms. This work extends propositions to the design and doped with europium of ionothermal synthesis MOFs, and explores their potential applications in the field of fluorescence sensing.

A new multifunctional phenanthroline-derived probe for colorimetric sensing of Fe2+ and fluorometric sensing of Zn2.

A new phenanthroline derivativebearing imidazole group, (2-(3,5-di(pyridin-4-yl)phenyl)-1-p-tolyl-1H-imidazo[4,5-f][1,10]phenanthroline) (1), has been devised. 1can be used as a multifunctionalprobe exhibiting a highly sensitive colorimetric response to Fe2+ and a selectively ratiometric fluorescent response to Zn2+in a buffer-ethanol solution. The absorption enhancement accompanied by a visual color change from colorless to red upon addition of Fe2+, makes 1a suitable naked-eye sensor for Fe2+. Moreover, 1displayed a Zn2+-induced red-shift of emission (44 nm) showing a color change from blue to light cyan under a 365-nm UV lamp. Its practical imaging applicability for intracellular Zn2+was confirmed in HeLa cells using a confocal microscope. The improved emission properties and cell imaging capability would provideanew approach forfluorescencesensationfor Zn2+.

Multifunctional lipid droplet probes for observing the polarity change of ferroptosis, inflammation, fatty liver and evaluating the efficacy of drugs

BackgroundLipid droplets (LDs) polarity is intricately linked to diverse biological processes and diseases. The visualization of LDs-polarity is of vital importance but challenging due to the lack of high-specificity, high-sensitivity and large-Stokes shift probes for real-time tracking LDs-polarity in biological systems. ResultsFour D-π-A based fluorescent probes (TPA-TCF1-TPA-TCF4) have been developed by combining tricyanofuran (an electron acceptor, A) and triphenylamine (an electron donor, D) derivatives with different terminal groups. Among them, TPA-TCF1 and TPA-TCF4 exhibit excellent polar sensitivity, large Stokes shift (≥182 nm in H2O), and efficient LDs targeting ability. In particular, TPA-TCF4 is capable of monitoring the change of LDs-polarity during ferroptosis, inflammation, apoptosis of cancer cell, and fatty liver. SignificanceAll these features render TPA-TCF4 a versatile tool for pharmacodynamic evaluation of anti-cancer drugs, in-depth understanding of the biological effect of LDs on ferroptosis, and medical diagnosis of LDs-polarity related diseases.