Using 2-dicyanomethylene-3-cyano-4,5,5-trimethyl-2,5-dihydrofuran(TCF)as a near-infrared fluorescent chromophore,we designed and synthesized a TCF-based fluorescent probe TCF-NS by introducing 2,4-dinitrophenyl ether ...Using 2-dicyanomethylene-3-cyano-4,5,5-trimethyl-2,5-dihydrofuran(TCF)as a near-infrared fluorescent chromophore,we designed and synthesized a TCF-based fluorescent probe TCF-NS by introducing 2,4-dinitrophenyl ether as the recognized site for H_(2)S.The probe TCF-NS displayed a rapid-response fluorescent against H_(2)S with high sensitivity and selection but had no significant fluorescence response to other biothiols.Furthermore,TCF-NS was applied to sense H_(2)S in living cells successfully with minimized cytotoxicity and a large Stokes shift.展开更多
Gallstones are a common disease worldwide,often leading to obstruction and inflammatory complications,which seriously affect the quality of life of patients.Research has shown that gallstone disease is associated with...Gallstones are a common disease worldwide,often leading to obstruction and inflammatory complications,which seriously affect the quality of life of patients.Research has shown that gallstone disease is associated with ferroptosis,lipid droplets(LDs),and abnormal levels of nitric oxide(NO).Fluorescent probes provide a sensitive and convenient method for detecting important substances in life systems and diseases.However,so far,no fluorescent probes for NO and LDs in gallstone disease have been reported.In this work,an effective ratiometric fluorescent probe LR-NH was designed for the detection of NO in LDs.With an anthracimide fluorophore and a secondary amine as a response site for NO,LR-NH exhibits high selectivity,sensitivity,and attractive ratiometric capability in detecting NO.Importantly,it can target LDs and shows excellent imaging ability for NO in cells and ferroptosis.Moreover,LR-NH can target the gallbladder and image NO in gallstone disease models,providing a unique and unprecedented tool for studying NO in LDs and gallbladder.展开更多
Ferroptosis is a new regulated cell death process executed by lipid peroxidation(LPO)of polyunsaturated fatty acids.Lipid droplets(LDs),as an important organelle for lipid storage and metabolism,are probably a major s...Ferroptosis is a new regulated cell death process executed by lipid peroxidation(LPO)of polyunsaturated fatty acids.Lipid droplets(LDs),as an important organelle for lipid storage and metabolism,are probably a major site of LPO and play critical roles in the regulation of ferroptosis.However,the detailed study on LPO in LDs has not been carried out because of the lack of LD-targeting tools for the in situ monitoring of LPO.Herein,the first LD-targeting LPO fluorescence probe(LD-LPO)has been developed.LD-LPO exhibits a rapid and selective fluorescence enhancement at 518 nm,which is unaffected by highly destructive reactive oxygen species(e.g.,hydroxyl radical)and environmental factor changes(e.g.,polarity and viscosity).LD-LPO is capable of targeting LDs and visualizing LPO within LDs in situ during erastin-or(1S,3R)-RSL3(RSL3)-induced ferroptosis.Moreover,LD-LPO has also been used to image LPO in the ferroptosis-associated non-alcoholic fatty liver disease(NAFLD),and to evaluate the medicine treatment of NAFLD with saroglitazar,demonstrating its utility for monitoring LPO levels in biosystems.The favorable analytical and imaging performance of LD-LPO may allow its application in more ferroptosisassociated physiological and pathological processes.展开更多
Fluorescent probes have wide applications in biological and environmental analysis due to their advantages of simple operation, convenient flexibility, high sensitivity and efficiency. They are considered to be promis...Fluorescent probes have wide applications in biological and environmental analysis due to their advantages of simple operation, convenient flexibility, high sensitivity and efficiency. They are considered to be promising tools for accurate analysis of agriculture- and food-related hazardous substances. In this review, the types and characteristics of the near-infrared fluorescence probes (NIFPs) are briefly described. The recent advances of NIFPs for precisely detecting various hazardous substances including heavy metals, sulfite and related sulfiting agents and hydrogen peroxide are summarized. Finally, the present challenges and future perspectives faced by NIFPs in food safety analysis are discussed.展开更多
A ratiometric fluorescent probe for hypoxanthine(Hx)detection was established based on the mimic enzyme and fluorescence characteristics of cobalt-doped graphite-phase carbon nitride(Co doped g-C_(3)N_(4)).In addition...A ratiometric fluorescent probe for hypoxanthine(Hx)detection was established based on the mimic enzyme and fluorescence characteristics of cobalt-doped graphite-phase carbon nitride(Co doped g-C_(3)N_(4)).In addition to emitting strong fluorescence,the peroxidase activity of Co doped g-C_(3)N_(4)can catalyze the reaction of O-phenylenediamine and H_(2)O_(2)to produce diallyl phthalate which can emit yellow fluorescence at 570 nm.Through the decomposition of Hx by xanthine oxidase,Hx can be indirectly detected by the generating hydrogen peroxide based on the measurement of fluorescent ratio I(F_(570)/F_(370)).The linear range was 1.7-272.2 mg/kg(R^(2)=0.997),and the detection limit was 1.52 mg/kg(3σ/K,n=9).The established method was applied to Hx detection in bass,grass carp,and shrimp,and the data were verified by HPLC.The result shows that the established probe is sensitive,accurate,and reliable,and can be used for Hx detection in aquatic products.展开更多
Humanβ-galactosidase(β-gal)is recognized as a crucial biomarker for evaluating senescence at the cellular and tissue levels in humans.However,tools to precisely track the endogenousβ-gal are still limited.Herein,we...Humanβ-galactosidase(β-gal)is recognized as a crucial biomarker for evaluating senescence at the cellular and tissue levels in humans.However,tools to precisely track the endogenousβ-gal are still limited.Herein,we present two novel self-calibratingβ-gal probes 7a and 7b which were constructed on a unique green/red dual-emissive fluorescence platform.The two probes inherently exhibited a stable green fluorescence signal impervious toβ-gal activity,serving as a reliable internal reference.They also displayed a progressively diminishing red fluorescence signal with the increasing ofβ-gal expression levels.The dual behavior endows them with self-calibration capacity and then renders excellently selective and sensitive for precisely monitoringβ-gal activity.Notably,compared with E.coliβ-gal,the two probes are more effectively response to A.oryzaeβ-gal homologous to humanβ-gal,indicating their unique species-selectivity.Furthermore,7a was validated for its effectiveness in determining senescenceassociatedβ-galactosidase(SA-β-gal)expression in senescent NRK-52E and HepG2 cells,underscoring its practical applicability in senescence research.展开更多
To address the lack of systematic studies on heavy metal fluorescent probes in typical buffer solutions,this study developed a Fe^(3+)and Cu^(2+)fluorescent probe,DHU‑NP‑4,based on a naphthalimide fluorophore.Comparat...To address the lack of systematic studies on heavy metal fluorescent probes in typical buffer solutions,this study developed a Fe^(3+)and Cu^(2+)fluorescent probe,DHU‑NP‑4,based on a naphthalimide fluorophore.Comparative analysis of the probe's performance in various buffer systems revealed that buffers with high organic content are unsuitable for evaluating such probes.Furthermore,the pH of the solvent system was found to significantly influence the probe's behavior.Under highly acidic conditions(pH≤2),DHU‑NP‑4 exhibited exceptional specificity for Fe^(3+),while in alkaline conditions,it demonstrated high specificity for Cu^(2+).Leveraging these properties,the probe enabled the quantitative detection of Fe^(3+)and Cu^(2+)in solution.展开更多
The presence of aluminum(Al^(3+))and fluoride(F^(−))ions in the environment can be harmful to ecosystems and human health,highlighting the need for accurate and efficient monitoring.In this paper,an innovative approac...The presence of aluminum(Al^(3+))and fluoride(F^(−))ions in the environment can be harmful to ecosystems and human health,highlighting the need for accurate and efficient monitoring.In this paper,an innovative approach is presented that leverages the power of machine learning to enhance the accuracy and efficiency of fluorescence-based detection for sequential quantitative analysis of aluminum(Al^(3+))and fluoride(F^(−))ions in aqueous solutions.The proposed method involves the synthesis of sulfur-functionalized carbon dots(C-dots)as fluorescence probes,with fluorescence enhancement upon interaction with Al^(3+)ions,achieving a detection limit of 4.2 nmol/L.Subsequently,in the presence of F^(−)ions,fluorescence is quenched,with a detection limit of 47.6 nmol/L.The fingerprints of fluorescence images are extracted using a cross-platform computer vision library in Python,followed by data preprocessing.Subsequently,the fingerprint data is subjected to cluster analysis using the K-means model from machine learning,and the average Silhouette Coefficient indicates excellent model performance.Finally,a regression analysis based on the principal component analysis method is employed to achieve more precise quantitative analysis of aluminum and fluoride ions.The results demonstrate that the developed model excels in terms of accuracy and sensitivity.This groundbreaking model not only showcases exceptional performance but also addresses the urgent need for effective environmental monitoring and risk assessment,making it a valuable tool for safeguarding our ecosystems and public health.展开更多
Colorectal cancer(CRC)is one of the most prevalent malignant tumors worldwide,exhibiting high morbidity and mortality.Lack of efficient tools for early diagnosis and surgical resection guidance of CRC have been a seri...Colorectal cancer(CRC)is one of the most prevalent malignant tumors worldwide,exhibiting high morbidity and mortality.Lack of efficient tools for early diagnosis and surgical resection guidance of CRC have been a serious threat to the long-term survival rate of the CRC patients.Recent studies have shown that relative higher viscosity was presented in tumor cells compared to that in normal cells,leading to viscosity as a potential biomarker for CRC.Herein,we reported the development of a series of novel viscosity-sensitive and mitochondria-specific fluorescent probes(HTB,HTI,and HTP)for CRC detection.Among them,HTB showed high sensitivity,minimal background interference,low cytotoxicity,and significant viscous response capability,making it an ideal tool for distinguishing colorectal tumor cells from normal cells.Importantly,we have successfully utilized HTB to visualize in a CRC-cells-derived xenograft(CDX)model,enriching its medical imaging capacity,which laid a foundation for further clinical translational application.展开更多
Rheumatoid arthritis(RA) is a chronic inflammatory disease with multi-system damage and autoimmune features.The main clinical manifestations of RA include joint pain,swelling,and stiffness,and RA may lead to joint def...Rheumatoid arthritis(RA) is a chronic inflammatory disease with multi-system damage and autoimmune features.The main clinical manifestations of RA include joint pain,swelling,and stiffness,and RA may lead to joint deformity and dysfunction in severe cases.The pathologic development of RA involves complex interactions of multiple biomarkers,and detecting a single biomarker may produce falsepositive results due to other confounding factors.Therefore,fluorescent probes that can detect multiple biomarkers simultaneously are crucial for precise RA diagnosis.Peroxynitrite(ONOO^(-)) and viscosity are inflammation-related factors in cells.In this study,we developed a dual responsive near-infrared fluorescent probe,YLS,for ONOO^(-) and viscosity.The probe features dual-channel turn-on fluorescence responses at 625 and 760 nm upon the presence of ONOO^(-) and viscosity,respectively.Supported by YLS,we found that during RA pathology,lymphocyte infiltration not only increases the concentration of proteins in the joint fluid resulting in elevated viscosity;at the same time,the overproduction of ONOO^(-) exacerbates oxidative stress and inflammatory responses.This multiparameter assay is expected to improve the diagnostic accuracy of the early stages of RA,thus providing a scientific basis for early intervention and personalized treatment.展开更多
In this study,a self-calibrating near-infrared fluorescence probe was designed and synthesized based on the dual-fluorophore strategy utilizing methylene blue and coumarin.The probe utilized methylene blue(emission sp...In this study,a self-calibrating near-infrared fluorescence probe was designed and synthesized based on the dual-fluorophore strategy utilizing methylene blue and coumarin.The probe utilized methylene blue(emission spectrum range:640-740 nm)and coumarin fluorophore(emission spectrum range:440-600 nm)as signal output units,thereby achieving effective spectral separation and highly selective detection of HClO.Under physiological pH conditions,HClO triggers an oxidation-cleavage reaction,releasing methylene blue and coumarin,which emit distinct red and green fluorescence,respectively.This dual-emission feature enabled rapid HClO detection with two-channel detection limits of 25.13 nmol·L^(-1)(green channel)and 31.55 nmol·L^(-1)(red channel).Furthermore,in cell imaging experiments,this probe demonstrated excellent cell membrane permeability and low cytotoxicity,successfully enabling the monitoring of both endogenous and exogenous HClO in living cells.By incorporating a twochannel self-calibration system,the probe effectively mitigated signal variations caused by instrumental or environmental interference,substantially improving detection sensitivity and reliability.展开更多
The abnormal metabolic activity of the tumor can increase the oxygen consumption in tumor cells,and the poor blood perfusion often happens in tumor regions as well,which are the main reasons that result in a hypoxic s...The abnormal metabolic activity of the tumor can increase the oxygen consumption in tumor cells,and the poor blood perfusion often happens in tumor regions as well,which are the main reasons that result in a hypoxic situation in the tumor.A fluorescence probe,AQD,with selective response toward hypoxia was designed for the detection of hypoxic tumor cells,which was obtained by the covalent connection of a large planar conjugated fluorophore with good fluorescence stability and a N,N-dimethylaniline moiety via the azo bond.The introduction of the azo bond in AQD caused significant fluorescence emission quenching,and the probe was reduced under hypoxic conditions to release the fluorophore via breaking the azo bond,resulting in the gradual recovery of fluorescence emission.Probe AQD exhibited a remarkable fluorescence response in hypoxic conditions,high selectivity,and good biocompatibility,which was successfully used for the imaging of hypoxic tumor cells and realized the detection of hypoxic A549 cells.展开更多
Herein,a luminescent europium-based metal-organic framework(Eu-MOF,[Eu_(3)(L)(HL)(NO_(3))_(2)(DMF)_(2)]·4DMF·5H_(2)O,H_(4)L=5,5′-(pyrazine-2,6-diyl)diisophthalic acid,DMF=N,N-dimethylformamide)was developed...Herein,a luminescent europium-based metal-organic framework(Eu-MOF,[Eu_(3)(L)(HL)(NO_(3))_(2)(DMF)_(2)]·4DMF·5H_(2)O,H_(4)L=5,5′-(pyrazine-2,6-diyl)diisophthalic acid,DMF=N,N-dimethylformamide)was developed for the dual-functional detection of environmental pollutants.This fluorescence-quenching-based sensor exhibited excep-tional sensitivity for both 2,4,6-trinitrophenol(TNP)and tetracycline(TC),achieving remarkably low detection lim-its of 1.96×10^(-6)and 1.71×10^(-7)mol·L^(-1),respectively.Notably,the system exhibited 99%fluorescence quenching ef-ficiency for TC,indicating ultra-efficient analyte recognition.The detection performance surpasses most reported lu-minescent MOF sensors,attributed to synergistic mechanisms of fluorescence resonance energy transfer(FRET)and photoinduced electron transfer(PET).CCDC:2446483.展开更多
A zinc sulfate open framework matrix,[Zn(SO_4)(DMSO)](1),was synthesized by solvothermal evaporationusing dimethyl sulfoxide(DMSO)as the solvent.A compositeP@1,which exhibits fluorescence and room tempera-ture phospho...A zinc sulfate open framework matrix,[Zn(SO_4)(DMSO)](1),was synthesized by solvothermal evaporationusing dimethyl sulfoxide(DMSO)as the solvent.A compositeP@1,which exhibits fluorescence and room tempera-ture phosphorescence(RTP)properties,was prepared by doping 2,6-naphthalic acid(P)into matrix1at a low con-centration.P@1emitted a green RTP that was visible to the naked eye and lasted for approximately 2 s.P@1exhib-ited selective phosphorescence enhancement response towards Pb^(2+),with a detection limit of 2.52μmol·L^(-1).Themain detection mechanism is the Pb—O coordination-induced phosphorescence enhancement in the system.Inter-estingly,P@1also functioned as a dual-channel probe for the rapid detection of Fe^(3+)ions through fluorescencequenching with a detection limit of 0.038μmol·L^(-1).The recognition mechanism may be attributed to the competi-tive energy absorption betweenP@1and Fe^(3+)ions.CCDC:2388502,1.展开更多
Excited-state intramolecular proton-transfer(ESIPT)based fluorescence probes are particularly attractive due to their unique properties including environmental sensitivity,a large Stokes shift,and potential for ratiom...Excited-state intramolecular proton-transfer(ESIPT)based fluorescence probes are particularly attractive due to their unique properties including environmental sensitivity,a large Stokes shift,and potential for ratiometric sensing.In general,ESIPT-based fluorophore incorporates an intramolecular hydrogen bonding interaction between a hydrogen bond donor(-OH and NH_(2)are common)and a hydrogen bond acceptor(C=N and C=O).More,protection-deprotection of hydroxyl group as hydrogen bond donor could induce an off-on switch of ESIPT-based emission.Therefore,protection-deprotection of hydroxyl group has been the widely used strategy to design fluorescent probes,where the potential key issue is selecting a protective group that can specifically leave in the presence of the target analyte.In this review,we mainly summarize the specific protecting groups(sites)and deprotection mechanisms for biologically important species(including reactive sulfur species(RSS),reactive oxygen species(ROS),enzymes,etc.),and analyze the advantages and disadvantages of different protection mechanisms from some aspects including probe stability,selectivity,response rate and assay system,etc.Based on the aforementioned,we further point out the current challenges and the potential future direction for developing ESIPT-based probes.展开更多
Early recognition is key to improving the prognosis of ischemic stroke(IS),while available imaging methods tend to target events that have already undergone ischemia.A new method to detect early IS is urgently needed,...Early recognition is key to improving the prognosis of ischemic stroke(IS),while available imaging methods tend to target events that have already undergone ischemia.A new method to detect early IS is urgently needed,as well as further study of its mechanisms.Viscosity and cysteine(Cys)levels of mitochondria have been associated with ferroptosis and IS.It is possible to identify IS and ferroptosis accurately and early by monitoring changes in mitochondrial Cys and viscosity simultaneously.In this work,a viscosity/Cys dual-responsive mitochondrial-targeted near-infrared(NIR)fluorescent probe(NVCP)was constructed for the precise tracking of IS using a two-dimensional design strategy.NVCP consists of a chromophore dyad containing diethylaminostyrene quinolinium rotor and chloro-sulfonylbenzoxadiazole(SBD-Cl)derivative with two easily distinguished emission bands(λ_(em)=592 and 670 nm).NVCP performs the way of killing two birds with one stone,that is,the probe exhibits excellent selectivity and sensitivity for detecting viscosity and Cys in living cells with excellent biocompatibility and accurate mitochondrial targeting capability by dual channel imaging mode.In addition,NVCP recognized that the viscosity increases and Cys level decreases in cells when undergoing ferroptosis and oxygen-glucose deprivation(OGD)stress by confocal imaging,flow cytometry,and Western blot experiments.Treatment of ferroptosis inhibitors(ferrostatin-1(Fer-1)and deferoxamine(DFO))could reverse the variation tendency of viscosity and Cys.This is the first time that the relationship between ferroptosis and IS was identified through an analysis of Cys and viscosity.More importantly,the ischemic area was also instantly distinguished from normal tissues through fluorescence imaging of NVCP in vivo.The developed NIR dual-responsive probe NVCP toward viscosity and Cys could serve as a sensitive and reliable tool for tracking ferroptosis-related pathological processes during IS.展开更多
Hypochlorous acid(HClO)is a critical biomolecule in living organisms,playing an essential role in numerous physiological or pathological processes.Abnormal levels of HClO in the body may lead to a series of diseases,f...Hypochlorous acid(HClO)is a critical biomolecule in living organisms,playing an essential role in numerous physiological or pathological processes.Abnormal levels of HClO in the body may lead to a series of diseases,for instance,inflammation and cancer.Thus,accurate measurement of HClO levels should be more beneficial for understanding its role in diseases and gaining a deeper insight into the pathogenesis of diseases.In this work,we designed a near-infrared two-photon fluorescent probe(HDM-Cl-HClO)for detecting fluctuations in HClO levels in inflammatory and tumor-bearing mice.Notably,the probe can respond to HClO within 5 s and trigger a brilliant red fluorescence at 660 nm.It exhibits high specificity and sensitivity for HClO.The superior spectral capability of the probe has enabled the detection of HClO levels in cells and zebrafish,as well as achieved the detection of HClO in inflammatory and tumor mice.This work not only provides a novel strategy and tool for HClO imaging in living systems,but also holds great potential for the diagnosis of inflammation and cancer.展开更多
Singlet oxygen(^(1)O_(2)),as the primary reactive oxygen species in photodynamic therapy,can effectively induce excessive oxidative stress to ablate tumors and kill germs in clinical treatment.However,monitoring endog...Singlet oxygen(^(1)O_(2)),as the primary reactive oxygen species in photodynamic therapy,can effectively induce excessive oxidative stress to ablate tumors and kill germs in clinical treatment.However,monitoring endogenous^(1)O_(2)is greatly challenging due to its extremely short lifetime and high reactivity in biological condition.Herein,we report an ultra-high signal-to-ratio near-infrared chemiluminescent probe(DCMCy)for the precise detection of endogenous^(1)O_(2)during photodynamic therapy(PDT).The methoxy moiety was removed from enolether unit in DCM-Cy to suppress the potential self-photooxidation reaction,thus greatly eliminating the photoinduced background signals during PDT.Additionally,the compact cyclobutane modification of DCM-Cy resulted in a significant 6-fold increase in cell permeability compared to conventional adamantane-dioxane probes.Therefore,our“step-by-step”strategy for DCM-Cy addressed the limitations of traditional chemiluminescent(CL)probes for^(1)O_(2),enabling effectively tracking of endogenous^(1)O_(2)level changes in living cells,pathogenic bacteria and mice in PDT.展开更多
Fluorescence-based imaging applications have been benefiting greatly from donor-acceptor(D-A)/donor-π-acceptor(D-π-A)fluorescent probes owing to their intramolecular charge transfer(ICT)nature and self-assembly beha...Fluorescence-based imaging applications have been benefiting greatly from donor-acceptor(D-A)/donor-π-acceptor(D-π-A)fluorescent probes owing to their intramolecular charge transfer(ICT)nature and self-assembly behavior.In this study,we design and synthesize a hydrophilic D-A fluorescent probe,namely CHBA,which would self-assemble into interlaced textures down to nanoscale but disassemble by trace amount of water in fingertip area.Upon finger-pressing,it enables fingerprint imaging and covers level-1/2/3 fingerprint information,wherein the sweat pores can be mapped in both bright field model and fluorescence mode,capable of naked-eye-based similarity analysis for personal identity verification(PIV).Spectroscopic analysis and morphology study show that the working mechanism can be attributed to the selective water-erosion effect on the solid-liquid interphase under physical contact.The sweat pore information can be digitized by polar coordinate conversion,further allowing machine-learning-based analysis for PIV application.The final PIV accuracy reaches 100%for all the involved machine-learning models,with no erroneous judgements.A prototype of PIV system is constructed by integrating CHBA with artificial intelligence hardware,wherein the sweat pore imaging,data processing and the decisionmaking could be run in parallel,suggesting high feasibility in real-world application.展开更多
The cell membrane,a fluid interface composed of self-assembled phospholipid molecules,is a vital component of biological systems that maintains cellular stability and prevents the invasion of foreign toxins.Due to its...The cell membrane,a fluid interface composed of self-assembled phospholipid molecules,is a vital component of biological systems that maintains cellular stability and prevents the invasion of foreign toxins.Due to its inherent fluidity,the cell membrane can undergo bending,shearing,and stretching,making membrane deformation crucial in processes like cell adhesion,migration,phagocytosis,and signal transduction.Within the plasma membrane are highly ordered dynamic structures formed by lipid molecules,known as“lipid rafts,”whose dynamic dissociation and reorganization are prerequisites for membrane deformation.Fluorescent probes have emerged as vital tools for studying these dynamic processes,offering a non-destructive,in situ,and real-time imaging method.By strategically designing these probes,researchers can image not only the microdomains of cell membranes but also explore more complex processes such as membrane fusion and fission.This review systematically summarizes the latest advancements in the application of fluorescent probes for cell membrane imaging.It also discusses the current challenges and provides insights into future research directions.We hope this review inspires further studies on the dynamic processes of complex cell membranes using fluorescent probes,ultimately advancing our understanding of the mechanisms underlying membrane dissociation,reorganization,fusion,and separation,and fostering research and therapeutic development for membrane-associated diseases.展开更多
基金financially supported by the Natural Science Foundation of Jiangsu Province(Grant No.BK20241181)the State Key Laboratory of AnalyticalChemistry for Life Science,School of Chemistry and Chemical Engineering,Nanjing University(Grant No.SKLACLS2419)。
文摘Using 2-dicyanomethylene-3-cyano-4,5,5-trimethyl-2,5-dihydrofuran(TCF)as a near-infrared fluorescent chromophore,we designed and synthesized a TCF-based fluorescent probe TCF-NS by introducing 2,4-dinitrophenyl ether as the recognized site for H_(2)S.The probe TCF-NS displayed a rapid-response fluorescent against H_(2)S with high sensitivity and selection but had no significant fluorescence response to other biothiols.Furthermore,TCF-NS was applied to sense H_(2)S in living cells successfully with minimized cytotoxicity and a large Stokes shift.
基金supported by the National Natural Science Foundation of China(No.22077044)the Natural Science Foundation of Hubei Province(No.2022CFA033).
文摘Gallstones are a common disease worldwide,often leading to obstruction and inflammatory complications,which seriously affect the quality of life of patients.Research has shown that gallstone disease is associated with ferroptosis,lipid droplets(LDs),and abnormal levels of nitric oxide(NO).Fluorescent probes provide a sensitive and convenient method for detecting important substances in life systems and diseases.However,so far,no fluorescent probes for NO and LDs in gallstone disease have been reported.In this work,an effective ratiometric fluorescent probe LR-NH was designed for the detection of NO in LDs.With an anthracimide fluorophore and a secondary amine as a response site for NO,LR-NH exhibits high selectivity,sensitivity,and attractive ratiometric capability in detecting NO.Importantly,it can target LDs and shows excellent imaging ability for NO in cells and ferroptosis.Moreover,LR-NH can target the gallbladder and image NO in gallstone disease models,providing a unique and unprecedented tool for studying NO in LDs and gallbladder.
基金the financial support from the National Natural Science Foundation of China(Nos.82060626,22004137,22164022,22174147,22074151,22374153,22174148)Excellent Youth scientific and technological talents of Guizhou Province(No.Qiankehe platform talents[2021]5638)+3 种基金Talents of Guizhou Science and Technology Cooperation Platform(No.[2020]4104)Science and Technology Innovation Team of Higher Education of Guizhou Provincial Education Department(No.Qianjiaoji[2023]073)Future Science and Technology Elite Talent Cultivation Project of Zunyi Medical University(No.ZYSE-2021-01)Zunyi Science and Technology Plan Project(No.Zunshi Keren Platform[2023]2)。
文摘Ferroptosis is a new regulated cell death process executed by lipid peroxidation(LPO)of polyunsaturated fatty acids.Lipid droplets(LDs),as an important organelle for lipid storage and metabolism,are probably a major site of LPO and play critical roles in the regulation of ferroptosis.However,the detailed study on LPO in LDs has not been carried out because of the lack of LD-targeting tools for the in situ monitoring of LPO.Herein,the first LD-targeting LPO fluorescence probe(LD-LPO)has been developed.LD-LPO exhibits a rapid and selective fluorescence enhancement at 518 nm,which is unaffected by highly destructive reactive oxygen species(e.g.,hydroxyl radical)and environmental factor changes(e.g.,polarity and viscosity).LD-LPO is capable of targeting LDs and visualizing LPO within LDs in situ during erastin-or(1S,3R)-RSL3(RSL3)-induced ferroptosis.Moreover,LD-LPO has also been used to image LPO in the ferroptosis-associated non-alcoholic fatty liver disease(NAFLD),and to evaluate the medicine treatment of NAFLD with saroglitazar,demonstrating its utility for monitoring LPO levels in biosystems.The favorable analytical and imaging performance of LD-LPO may allow its application in more ferroptosisassociated physiological and pathological processes.
基金supported by the National Natural Science Foundation of China (Nos. 81925019, 81801817 and U22A20333)the National Key Research and Development Program of China (Nos. 2023YFB3810000 and 2023YFB3810003)+1 种基金the Fundamental Research Funds for the Central Universities and the Fujian Basic Research Foundation (Nos. 2022J011403, 2023XAKJ0101009, B2302014 and 2020Y4003)the Program for New Century Excellent Talents in University, China (No. NCET-13-0502).
文摘Fluorescent probes have wide applications in biological and environmental analysis due to their advantages of simple operation, convenient flexibility, high sensitivity and efficiency. They are considered to be promising tools for accurate analysis of agriculture- and food-related hazardous substances. In this review, the types and characteristics of the near-infrared fluorescence probes (NIFPs) are briefly described. The recent advances of NIFPs for precisely detecting various hazardous substances including heavy metals, sulfite and related sulfiting agents and hydrogen peroxide are summarized. Finally, the present challenges and future perspectives faced by NIFPs in food safety analysis are discussed.
基金supported by the National Natural Science Foundation of China(21804050)the National Key R and D Program of China(2018YFD0901003)+2 种基金the Science and Technology Planning Project of Xiamen,China(3502Z20183031)the Fujian Provincial Fund Project(2018J01432)the Xiamen Science and Technology Planning Project,China(3502Z20183031)。
文摘A ratiometric fluorescent probe for hypoxanthine(Hx)detection was established based on the mimic enzyme and fluorescence characteristics of cobalt-doped graphite-phase carbon nitride(Co doped g-C_(3)N_(4)).In addition to emitting strong fluorescence,the peroxidase activity of Co doped g-C_(3)N_(4)can catalyze the reaction of O-phenylenediamine and H_(2)O_(2)to produce diallyl phthalate which can emit yellow fluorescence at 570 nm.Through the decomposition of Hx by xanthine oxidase,Hx can be indirectly detected by the generating hydrogen peroxide based on the measurement of fluorescent ratio I(F_(570)/F_(370)).The linear range was 1.7-272.2 mg/kg(R^(2)=0.997),and the detection limit was 1.52 mg/kg(3σ/K,n=9).The established method was applied to Hx detection in bass,grass carp,and shrimp,and the data were verified by HPLC.The result shows that the established probe is sensitive,accurate,and reliable,and can be used for Hx detection in aquatic products.
基金the financial support from the National Natural Science Foundation of China(Nos.21977082,22037002and 21472148)the Natural Science Basic Research Program of Shaanxi(No.2020JC-38)。
文摘Humanβ-galactosidase(β-gal)is recognized as a crucial biomarker for evaluating senescence at the cellular and tissue levels in humans.However,tools to precisely track the endogenousβ-gal are still limited.Herein,we present two novel self-calibratingβ-gal probes 7a and 7b which were constructed on a unique green/red dual-emissive fluorescence platform.The two probes inherently exhibited a stable green fluorescence signal impervious toβ-gal activity,serving as a reliable internal reference.They also displayed a progressively diminishing red fluorescence signal with the increasing ofβ-gal expression levels.The dual behavior endows them with self-calibration capacity and then renders excellently selective and sensitive for precisely monitoringβ-gal activity.Notably,compared with E.coliβ-gal,the two probes are more effectively response to A.oryzaeβ-gal homologous to humanβ-gal,indicating their unique species-selectivity.Furthermore,7a was validated for its effectiveness in determining senescenceassociatedβ-galactosidase(SA-β-gal)expression in senescent NRK-52E and HepG2 cells,underscoring its practical applicability in senescence research.
文摘To address the lack of systematic studies on heavy metal fluorescent probes in typical buffer solutions,this study developed a Fe^(3+)and Cu^(2+)fluorescent probe,DHU‑NP‑4,based on a naphthalimide fluorophore.Comparative analysis of the probe's performance in various buffer systems revealed that buffers with high organic content are unsuitable for evaluating such probes.Furthermore,the pH of the solvent system was found to significantly influence the probe's behavior.Under highly acidic conditions(pH≤2),DHU‑NP‑4 exhibited exceptional specificity for Fe^(3+),while in alkaline conditions,it demonstrated high specificity for Cu^(2+).Leveraging these properties,the probe enabled the quantitative detection of Fe^(3+)and Cu^(2+)in solution.
基金supported by the National Natural Science Foundation of China(No.U21A20290)Guangdong Basic and Applied Basic Research Foundation(No.2022A1515011656)+2 种基金the Projects of Talents Recruitment of GDUPT(No.2023rcyj1003)the 2022“Sail Plan”Project of Maoming Green Chemical Industry Research Institute(No.MMGCIRI2022YFJH-Y-024)Maoming Science and Technology Project(No.2023382).
文摘The presence of aluminum(Al^(3+))and fluoride(F^(−))ions in the environment can be harmful to ecosystems and human health,highlighting the need for accurate and efficient monitoring.In this paper,an innovative approach is presented that leverages the power of machine learning to enhance the accuracy and efficiency of fluorescence-based detection for sequential quantitative analysis of aluminum(Al^(3+))and fluoride(F^(−))ions in aqueous solutions.The proposed method involves the synthesis of sulfur-functionalized carbon dots(C-dots)as fluorescence probes,with fluorescence enhancement upon interaction with Al^(3+)ions,achieving a detection limit of 4.2 nmol/L.Subsequently,in the presence of F^(−)ions,fluorescence is quenched,with a detection limit of 47.6 nmol/L.The fingerprints of fluorescence images are extracted using a cross-platform computer vision library in Python,followed by data preprocessing.Subsequently,the fingerprint data is subjected to cluster analysis using the K-means model from machine learning,and the average Silhouette Coefficient indicates excellent model performance.Finally,a regression analysis based on the principal component analysis method is employed to achieve more precise quantitative analysis of aluminum and fluoride ions.The results demonstrate that the developed model excels in terms of accuracy and sensitivity.This groundbreaking model not only showcases exceptional performance but also addresses the urgent need for effective environmental monitoring and risk assessment,making it a valuable tool for safeguarding our ecosystems and public health.
基金supported by the National Natural Science Foundation of China(Nos.82272067,81974386,M-0696,and 82273486)Natural Science Foundation of Hunan Province(Nos.2022JJ80052,2024JJ6596)the Innovation Fund for Postgraduate Students of Central South University(No.2023ZZTS0841)。
文摘Colorectal cancer(CRC)is one of the most prevalent malignant tumors worldwide,exhibiting high morbidity and mortality.Lack of efficient tools for early diagnosis and surgical resection guidance of CRC have been a serious threat to the long-term survival rate of the CRC patients.Recent studies have shown that relative higher viscosity was presented in tumor cells compared to that in normal cells,leading to viscosity as a potential biomarker for CRC.Herein,we reported the development of a series of novel viscosity-sensitive and mitochondria-specific fluorescent probes(HTB,HTI,and HTP)for CRC detection.Among them,HTB showed high sensitivity,minimal background interference,low cytotoxicity,and significant viscous response capability,making it an ideal tool for distinguishing colorectal tumor cells from normal cells.Importantly,we have successfully utilized HTB to visualize in a CRC-cells-derived xenograft(CDX)model,enriching its medical imaging capacity,which laid a foundation for further clinical translational application.
基金the National Natural Science Foundation of China(Nos.22325703,22377071,U23A6009)Research Project Supported by Shanxi Scholarship Council of China(No.2022-002)+1 种基金the Shanxi Province Science Foundation(No.202203021221009)Shanxi Province Science and Technology activities for overseas people selected funding project(No.2024001)。
文摘Rheumatoid arthritis(RA) is a chronic inflammatory disease with multi-system damage and autoimmune features.The main clinical manifestations of RA include joint pain,swelling,and stiffness,and RA may lead to joint deformity and dysfunction in severe cases.The pathologic development of RA involves complex interactions of multiple biomarkers,and detecting a single biomarker may produce falsepositive results due to other confounding factors.Therefore,fluorescent probes that can detect multiple biomarkers simultaneously are crucial for precise RA diagnosis.Peroxynitrite(ONOO^(-)) and viscosity are inflammation-related factors in cells.In this study,we developed a dual responsive near-infrared fluorescent probe,YLS,for ONOO^(-) and viscosity.The probe features dual-channel turn-on fluorescence responses at 625 and 760 nm upon the presence of ONOO^(-) and viscosity,respectively.Supported by YLS,we found that during RA pathology,lymphocyte infiltration not only increases the concentration of proteins in the joint fluid resulting in elevated viscosity;at the same time,the overproduction of ONOO^(-) exacerbates oxidative stress and inflammatory responses.This multiparameter assay is expected to improve the diagnostic accuracy of the early stages of RA,thus providing a scientific basis for early intervention and personalized treatment.
文摘In this study,a self-calibrating near-infrared fluorescence probe was designed and synthesized based on the dual-fluorophore strategy utilizing methylene blue and coumarin.The probe utilized methylene blue(emission spectrum range:640-740 nm)and coumarin fluorophore(emission spectrum range:440-600 nm)as signal output units,thereby achieving effective spectral separation and highly selective detection of HClO.Under physiological pH conditions,HClO triggers an oxidation-cleavage reaction,releasing methylene blue and coumarin,which emit distinct red and green fluorescence,respectively.This dual-emission feature enabled rapid HClO detection with two-channel detection limits of 25.13 nmol·L^(-1)(green channel)and 31.55 nmol·L^(-1)(red channel).Furthermore,in cell imaging experiments,this probe demonstrated excellent cell membrane permeability and low cytotoxicity,successfully enabling the monitoring of both endogenous and exogenous HClO in living cells.By incorporating a twochannel self-calibration system,the probe effectively mitigated signal variations caused by instrumental or environmental interference,substantially improving detection sensitivity and reliability.
文摘The abnormal metabolic activity of the tumor can increase the oxygen consumption in tumor cells,and the poor blood perfusion often happens in tumor regions as well,which are the main reasons that result in a hypoxic situation in the tumor.A fluorescence probe,AQD,with selective response toward hypoxia was designed for the detection of hypoxic tumor cells,which was obtained by the covalent connection of a large planar conjugated fluorophore with good fluorescence stability and a N,N-dimethylaniline moiety via the azo bond.The introduction of the azo bond in AQD caused significant fluorescence emission quenching,and the probe was reduced under hypoxic conditions to release the fluorophore via breaking the azo bond,resulting in the gradual recovery of fluorescence emission.Probe AQD exhibited a remarkable fluorescence response in hypoxic conditions,high selectivity,and good biocompatibility,which was successfully used for the imaging of hypoxic tumor cells and realized the detection of hypoxic A549 cells.
文摘Herein,a luminescent europium-based metal-organic framework(Eu-MOF,[Eu_(3)(L)(HL)(NO_(3))_(2)(DMF)_(2)]·4DMF·5H_(2)O,H_(4)L=5,5′-(pyrazine-2,6-diyl)diisophthalic acid,DMF=N,N-dimethylformamide)was developed for the dual-functional detection of environmental pollutants.This fluorescence-quenching-based sensor exhibited excep-tional sensitivity for both 2,4,6-trinitrophenol(TNP)and tetracycline(TC),achieving remarkably low detection lim-its of 1.96×10^(-6)and 1.71×10^(-7)mol·L^(-1),respectively.Notably,the system exhibited 99%fluorescence quenching ef-ficiency for TC,indicating ultra-efficient analyte recognition.The detection performance surpasses most reported lu-minescent MOF sensors,attributed to synergistic mechanisms of fluorescence resonance energy transfer(FRET)and photoinduced electron transfer(PET).CCDC:2446483.
文摘A zinc sulfate open framework matrix,[Zn(SO_4)(DMSO)](1),was synthesized by solvothermal evaporationusing dimethyl sulfoxide(DMSO)as the solvent.A compositeP@1,which exhibits fluorescence and room tempera-ture phosphorescence(RTP)properties,was prepared by doping 2,6-naphthalic acid(P)into matrix1at a low con-centration.P@1emitted a green RTP that was visible to the naked eye and lasted for approximately 2 s.P@1exhib-ited selective phosphorescence enhancement response towards Pb^(2+),with a detection limit of 2.52μmol·L^(-1).Themain detection mechanism is the Pb—O coordination-induced phosphorescence enhancement in the system.Inter-estingly,P@1also functioned as a dual-channel probe for the rapid detection of Fe^(3+)ions through fluorescencequenching with a detection limit of 0.038μmol·L^(-1).The recognition mechanism may be attributed to the competi-tive energy absorption betweenP@1and Fe^(3+)ions.CCDC:2388502,1.
基金National Natural Science Foundation of China(Nos.22277104,22325703,22074084)the Natural Science Foundation of Shanxi Province(No.202203021212184)+3 种基金Research Project supported by Shanxi Scholarship Council of China(No.2022-002)the Basic Research Program of Shanxi Province(Free Exploration)(No.202203021221009)2022 Lvliang City science and technology plan project(Nos.2022SHFZ51,2022GXYF15)Scientific Instrument Center of Shanxi University(No.201512)。
文摘Excited-state intramolecular proton-transfer(ESIPT)based fluorescence probes are particularly attractive due to their unique properties including environmental sensitivity,a large Stokes shift,and potential for ratiometric sensing.In general,ESIPT-based fluorophore incorporates an intramolecular hydrogen bonding interaction between a hydrogen bond donor(-OH and NH_(2)are common)and a hydrogen bond acceptor(C=N and C=O).More,protection-deprotection of hydroxyl group as hydrogen bond donor could induce an off-on switch of ESIPT-based emission.Therefore,protection-deprotection of hydroxyl group has been the widely used strategy to design fluorescent probes,where the potential key issue is selecting a protective group that can specifically leave in the presence of the target analyte.In this review,we mainly summarize the specific protecting groups(sites)and deprotection mechanisms for biologically important species(including reactive sulfur species(RSS),reactive oxygen species(ROS),enzymes,etc.),and analyze the advantages and disadvantages of different protection mechanisms from some aspects including probe stability,selectivity,response rate and assay system,etc.Based on the aforementioned,we further point out the current challenges and the potential future direction for developing ESIPT-based probes.
基金supported by The Natural Science Foundation of Hunan Province(Nos.2022JJ30484,2022JJ10042,2023JJ50157)The Start-up funds of University of South China(Nos.201RGC012,20XQD030)+3 种基金The Research Foundation of Education Bureau of Hunan Province(No.22B0418)Education Ministry’s Collaborative Education Program with Industry of China(Nos.220902102225257,230902999244208)Ministry of Education of China:“Chunhui Plan”(No.HZKY20220359)The Open Fund of State Key Laboratory of Chemo/Biosensing and Chemometrics(Nos.2021013,2021015).
文摘Early recognition is key to improving the prognosis of ischemic stroke(IS),while available imaging methods tend to target events that have already undergone ischemia.A new method to detect early IS is urgently needed,as well as further study of its mechanisms.Viscosity and cysteine(Cys)levels of mitochondria have been associated with ferroptosis and IS.It is possible to identify IS and ferroptosis accurately and early by monitoring changes in mitochondrial Cys and viscosity simultaneously.In this work,a viscosity/Cys dual-responsive mitochondrial-targeted near-infrared(NIR)fluorescent probe(NVCP)was constructed for the precise tracking of IS using a two-dimensional design strategy.NVCP consists of a chromophore dyad containing diethylaminostyrene quinolinium rotor and chloro-sulfonylbenzoxadiazole(SBD-Cl)derivative with two easily distinguished emission bands(λ_(em)=592 and 670 nm).NVCP performs the way of killing two birds with one stone,that is,the probe exhibits excellent selectivity and sensitivity for detecting viscosity and Cys in living cells with excellent biocompatibility and accurate mitochondrial targeting capability by dual channel imaging mode.In addition,NVCP recognized that the viscosity increases and Cys level decreases in cells when undergoing ferroptosis and oxygen-glucose deprivation(OGD)stress by confocal imaging,flow cytometry,and Western blot experiments.Treatment of ferroptosis inhibitors(ferrostatin-1(Fer-1)and deferoxamine(DFO))could reverse the variation tendency of viscosity and Cys.This is the first time that the relationship between ferroptosis and IS was identified through an analysis of Cys and viscosity.More importantly,the ischemic area was also instantly distinguished from normal tissues through fluorescence imaging of NVCP in vivo.The developed NIR dual-responsive probe NVCP toward viscosity and Cys could serve as a sensitive and reliable tool for tracking ferroptosis-related pathological processes during IS.
基金National Natural Science Foundation of China(No.22264013)Hainan Province Clinical Medical Center(No.2021)Hainan Province Science and Technology Special Fund(No.ZDYF2024SHFZ104).
文摘Hypochlorous acid(HClO)is a critical biomolecule in living organisms,playing an essential role in numerous physiological or pathological processes.Abnormal levels of HClO in the body may lead to a series of diseases,for instance,inflammation and cancer.Thus,accurate measurement of HClO levels should be more beneficial for understanding its role in diseases and gaining a deeper insight into the pathogenesis of diseases.In this work,we designed a near-infrared two-photon fluorescent probe(HDM-Cl-HClO)for detecting fluctuations in HClO levels in inflammatory and tumor-bearing mice.Notably,the probe can respond to HClO within 5 s and trigger a brilliant red fluorescence at 660 nm.It exhibits high specificity and sensitivity for HClO.The superior spectral capability of the probe has enabled the detection of HClO levels in cells and zebrafish,as well as achieved the detection of HClO in inflammatory and tumor mice.This work not only provides a novel strategy and tool for HClO imaging in living systems,but also holds great potential for the diagnosis of inflammation and cancer.
基金supported by National Natural Science Foundation of China(Nos.32121005,22225805,22308101,and 32394001)Shanghai Science and Technology Innovation Action Plan(No.23J21901600)+2 种基金Innovation Program of Shanghai Municipal Education Commission,Shanghai Frontier Science Research Base of Optogenetic Techniques for Cell Metabolism(Shanghai Municipal Education Commission,No.2021 Sci&Tech 03-28)the China Postdoctoral Science Foundation(No.2021M701199)Natural Science Foundation of Shanghai(No.23ZR1416600).
文摘Singlet oxygen(^(1)O_(2)),as the primary reactive oxygen species in photodynamic therapy,can effectively induce excessive oxidative stress to ablate tumors and kill germs in clinical treatment.However,monitoring endogenous^(1)O_(2)is greatly challenging due to its extremely short lifetime and high reactivity in biological condition.Herein,we report an ultra-high signal-to-ratio near-infrared chemiluminescent probe(DCMCy)for the precise detection of endogenous^(1)O_(2)during photodynamic therapy(PDT).The methoxy moiety was removed from enolether unit in DCM-Cy to suppress the potential self-photooxidation reaction,thus greatly eliminating the photoinduced background signals during PDT.Additionally,the compact cyclobutane modification of DCM-Cy resulted in a significant 6-fold increase in cell permeability compared to conventional adamantane-dioxane probes.Therefore,our“step-by-step”strategy for DCM-Cy addressed the limitations of traditional chemiluminescent(CL)probes for^(1)O_(2),enabling effectively tracking of endogenous^(1)O_(2)level changes in living cells,pathogenic bacteria and mice in PDT.
基金the financial support from National Natural Science Foundation of China(No.51703135)the technical support from Beijing Key Laboratory of Optical Materials and Photonic Devices。
文摘Fluorescence-based imaging applications have been benefiting greatly from donor-acceptor(D-A)/donor-π-acceptor(D-π-A)fluorescent probes owing to their intramolecular charge transfer(ICT)nature and self-assembly behavior.In this study,we design and synthesize a hydrophilic D-A fluorescent probe,namely CHBA,which would self-assemble into interlaced textures down to nanoscale but disassemble by trace amount of water in fingertip area.Upon finger-pressing,it enables fingerprint imaging and covers level-1/2/3 fingerprint information,wherein the sweat pores can be mapped in both bright field model and fluorescence mode,capable of naked-eye-based similarity analysis for personal identity verification(PIV).Spectroscopic analysis and morphology study show that the working mechanism can be attributed to the selective water-erosion effect on the solid-liquid interphase under physical contact.The sweat pore information can be digitized by polar coordinate conversion,further allowing machine-learning-based analysis for PIV application.The final PIV accuracy reaches 100%for all the involved machine-learning models,with no erroneous judgements.A prototype of PIV system is constructed by integrating CHBA with artificial intelligence hardware,wherein the sweat pore imaging,data processing and the decisionmaking could be run in parallel,suggesting high feasibility in real-world application.
基金supported by the National Nature Science Foundation of China(22107028)State Key Laboratory of Fine Chemicals,Dalian University of Technology(KF2307)+4 种基金Central Guidance Fund for Local Science and Technology Development Project(2024FRD05069)Natural Science Foundation of Chongqing(CSTB2023NSCQ-MSX0335)ML.wishes to thank the support of the National Natural Science Foundation of China(22308220)Shenzhen Uni-versity Third-Phase Project of Constructing High-Level University(000001032104)the Research Team Culti-vation Program of Shenzhen University(2023QNT005).
文摘The cell membrane,a fluid interface composed of self-assembled phospholipid molecules,is a vital component of biological systems that maintains cellular stability and prevents the invasion of foreign toxins.Due to its inherent fluidity,the cell membrane can undergo bending,shearing,and stretching,making membrane deformation crucial in processes like cell adhesion,migration,phagocytosis,and signal transduction.Within the plasma membrane are highly ordered dynamic structures formed by lipid molecules,known as“lipid rafts,”whose dynamic dissociation and reorganization are prerequisites for membrane deformation.Fluorescent probes have emerged as vital tools for studying these dynamic processes,offering a non-destructive,in situ,and real-time imaging method.By strategically designing these probes,researchers can image not only the microdomains of cell membranes but also explore more complex processes such as membrane fusion and fission.This review systematically summarizes the latest advancements in the application of fluorescent probes for cell membrane imaging.It also discusses the current challenges and provides insights into future research directions.We hope this review inspires further studies on the dynamic processes of complex cell membranes using fluorescent probes,ultimately advancing our understanding of the mechanisms underlying membrane dissociation,reorganization,fusion,and separation,and fostering research and therapeutic development for membrane-associated diseases.