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 rapid development of super-resolution microscopy has made it possible to observe subcellular structures and dynamic behaviors in living cells with nanoscale spatial resolution, greatly advancing progress in life s...The rapid development of super-resolution microscopy has made it possible to observe subcellular structures and dynamic behaviors in living cells with nanoscale spatial resolution, greatly advancing progress in life sciences. As hardware technology continues to evolve, the availability of new fluorescent probes with superior performance is becoming increasingly important. In recent years, fluorescent nanoprobes (FNPs) have emerged as highly promising fluorescent probes for bioimaging due to their high brightness and excellent photostability. This paper focuses on the development and applications of FNPs as probes for live-cell super-resolution imaging. It provides an overview of different super-resolution methods, discusses the performance requirements for FNPs in these methods, and reviews the latest applications of FNPs in the super-resolution imaging of living cells. Finally, it addresses the challenges and future outlook in this field.展开更多
PBQ[1-(4-chlorophenyl)-3-(pyridin-3-yl)urea],an enormous potent molluscicide,showed excellent Pomacea canaliculata(P.canaliculata)control activity and low toxicity for other aquatic organisms,but its snail-killing mec...PBQ[1-(4-chlorophenyl)-3-(pyridin-3-yl)urea],an enormous potent molluscicide,showed excellent Pomacea canaliculata(P.canaliculata)control activity and low toxicity for other aquatic organisms,but its snail-killing mechanisms are still not fully understood.We employed an optical method to elucidate PBQ action via a novel fluorescent viscosity probe,NCV.As the viscosity in the test solutions increased,compared with that in pure ethanol,a 54-fold fluorescence intensity enhancement of NCV was observed in 310 cP of 90%glycerol.Furthermore,NCV successfully exhibited a selective fluorescence response towards monensin-induced cellular viscosity changes in HepG2 cells.The liver,stomach,and foot plantar of the tested snails were frozen and sectioned for fluorescent imaging experiments after the treatment with different PBQ concentrations over various times.A significant fluorescent increase in the snail's liver was observed upon exposure to 0.75 mg/L PBQ for 72 h,which highlighted an increase in viscosity.Hematoxylin and eosin(HE)staining further supported PBQ-induced liver damage with a viscosity increase in P.canaliculata.Our study provides a new rapid optical visualization method to study the killing mechanisms of PBQ and may help discover new chemicals that control snail populations.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
The fluorescence imaging (FLI) in the second near-infrared window (NIR-II, 1000–1700nm) has attracted considerable attention in the past decade. In contrast to conventional NIR-I window excitation (808nm/980nm), FLI ...The fluorescence imaging (FLI) in the second near-infrared window (NIR-II, 1000–1700nm) has attracted considerable attention in the past decade. In contrast to conventional NIR-I window excitation (808nm/980nm), FLI with NIR-II window excitation (1064nm/other wavelength beyond 1000nm) can afford deeper tissue penetration depth with high clarity due to the merits of suppressed photon scattering and diminished autofluorescence. In this review, we have summarized NIR-II window excitable/emissive organic/polymeric fluorophores recently developed. The characteristics of these fluorophores such as chemical structures and photophysical properties have also been critically discussed. Furthermore, the latest development of noninvasive in vivo FLI with NIR-II excitation was highlighted. The ideal imaging results emphasized the importance of NIR-II excitation of these fluorophores in enabling deep tissue penetration and high-resolution imaging. Finally, a perspective on the challenges and prospects of NIR-II excitable/emissive organic/polymeric fluorophores was also discussed. We expected this review will be served as a source of inspiration for researchers, stimulating the creation of novel NIR-II excitable fluorophores and fostering the development of bioimaging applications.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
We report five coordination polymers(CPs)based on fluorescent ligands[1,6-di(1H-imidazol-1-yl)pyrene(dip),9,10-di(1H-imidazol-1-yl)anthracene(dia)]and anionic ligands[cyclohexane-1,4-dicarboxylic acid(H_(2)cda),campho...We report five coordination polymers(CPs)based on fluorescent ligands[1,6-di(1H-imidazol-1-yl)pyrene(dip),9,10-di(1H-imidazol-1-yl)anthracene(dia)]and anionic ligands[cyclohexane-1,4-dicarboxylic acid(H_(2)cda),camphoric acid(H_(2)cpa)].In[Cd(dip)(cda)]·4H_(2)O}_(n)(1),the Cd^(2+)ions,acting as tetrahedral nodes,are linked by dipand cda^(2-)ligands with four Cd^(2+)ions into five-fold interpenetrating network array of topology of dia.In{[Cd(dip)(cpa)]·4H_(2)O}_(n)(2),the Cd^(2+)ions,acting as a 4-connector,are linked by cpa^(2-)and dip ligands into a 3D framework ofcds topology.In{[Ni(dia)_(2)Cl_(2)]·DMF}_(n)(3),the Ni^(2+)ion is linked by four dia ligands into a layer structure,and 1Dchannels of a cross-section of 1.35 nm×0.96 nm are formed.In{[Cd(dia)_(2)(H_(2)O)_(2)](NO_(3))_(2)·2DMSO}n(4),the dia ligandsconnected Cd^(2+)ions into a 2D layer,and 1D channels are formed between adjacent layers with a cross-section of0.87 nm×0.43 nm.In[Zn(dip)Cl_(2)]_(n)(5),the Zn^(2+)ion is linked by dip ligands into an infinite 1D chain.The infrared,thermal gravimetric,and fluorescent emission data were collected and analyzed for these coordination polymers.CCDC:2356055,1;2440075,2;2356057,3;2356057,4;2356059,5.展开更多
Succinylcholine(SC)is a widely used depolarizing muscle relaxant,but improper use can lead to arrhythmias and,in severe cases,pose a life-threatening risk.Additionally,some criminals exploit SC for illicit activities....Succinylcholine(SC)is a widely used depolarizing muscle relaxant,but improper use can lead to arrhythmias and,in severe cases,pose a life-threatening risk.Additionally,some criminals exploit SC for illicit activities.Therefore,rapid SC detection is paramount for clinical practice and public safety.Currently,however,limited methods are available for the rapid detection of SC.A fluorescent indicator displacement assay sensor based on molecular recognition of an amide naphthotube was developed.This sensor enabled the rapid fluorescent detection of SC through competitive binding between SC and methylene blue with the amide naphthotube.The sensor exhibited exceptional sensitivity with a detection limit as low as 1.1μmol/L and a detection range of 1.1~60μmol/L,coupled with outstanding selectivity and robust stability.Furthermore,this sensor accurately determined SC levels in biological samples such as serum.In summary,this research provides a new solution for the rapid and accurate sensing of SC in complex matrices and offers new insights for the swift identification and detection of toxins.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
Six coordination polymers based on 9,10-di(pyridine-4-yl)-anthracene(DPA)and 1,6-di(1H-imidazol-1-yl)pyrene(DIP)were obtained by solvothermal reactions.{[Zn(DPA)Cl_(2)]·DMF·2H_(2)O}n(1)and{[Zn_(1.5)(DPA)_(1....Six coordination polymers based on 9,10-di(pyridine-4-yl)-anthracene(DPA)and 1,6-di(1H-imidazol-1-yl)pyrene(DIP)were obtained by solvothermal reactions.{[Zn(DPA)Cl_(2)]·DMF·2H_(2)O}n(1)and{[Zn_(1.5)(DPA)_(1.5)Cl_(3)]·5H_(2)O}n(2)are framework isomers,which both contain zigzag chains formed by DPA,Zn^(2+),and Cl-.The zigzag chains in 1 are further assembled by C—H…Cl interactions into layers,and these layers exhibit two different orientations,displaying a rare 2D to 3D interpenetration mode.The zigzag chains in 2 are parallelly arranged.{[Zn_(3)(DPA)_(3)Br_(6)]·2DMF·_(1.5)H_(2)O}n(3)is isostructural to 2.3 was obtained using ZnBr_(2)instead of ZnCl_(2).[M(DPA)(formate)_(2)(H_(2)O)_(2)]n[M=Co(4),Cu(5)]are isostructural,contain chain structures formed by DPA,Cu^(2+)/Co^(2+),and for-mate ions,which were formed in situ in the solvothermal reaction.{[Zn(DIP)_(2)Cl]ClO_(4)}n(6)contains a layer structure formed by DIP and Zn^(2+).Free DPA and DIP ligands exhibited high fluorescence at room temperature,and coordina-tion polymers 3 and 6 displayed enhanced fluorescent emissions.展开更多
The rapid development of microfluidic technology has led to the evolution of microdroplets from simple emulsion structures to complex multilayered and multicompartmental configurations.These advancements have endowed ...The rapid development of microfluidic technology has led to the evolution of microdroplets from simple emulsion structures to complex multilayered and multicompartmental configurations.These advancements have endowed microdroplets with the capability to contain multiple compartments that remain isolated from one another,enabling them to carry different molecules of interest.Consequently,researchers can now investigate intricate spatially confined chemical reactions and signal transduction pathways within subcellular organelles.Moreover,modern microdroplets often possess excellent optical transparency,allowing fluorescently labelled,multi-layered,and compartmental droplets to provide detailed insights through real-time,in situ,and dynamic fluorescence imaging.Hence,this review systematically summarizes current methodologies for preparing multicomponent microdroplets and their applications,particularly focusing on fluorescent microdroplets.Additionally,it discusses existing critical challenges and outlines future research directions.By offering a comprehensive overview of the preparation methods and applications of fluorescent microdroplets,this review aims to stimulate the interest of researchers and foster their utilization in more complex and biomimetic environments.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
In recent years,cellulose-based fluorescent polymers have received considerable attention.However,conventional modification methods face challenges such as insolubility in most solvents,fluorescence instability,and en...In recent years,cellulose-based fluorescent polymers have received considerable attention.However,conventional modification methods face challenges such as insolubility in most solvents,fluorescence instability,and environmental risks.In this study,a novel biosynthesis strategy was developed to fabricate fluorescent cellulose by adding fluorescent glucose derivatives to a bacterial fermentation broth.The metabolic activity of bacteria is utilized to achieve in situ polymerization of glucose and its derivatives during the synthesis of bacterial cellulose.Owing to the structural similarity between triphenylamine-modified glucose(TPA-Glc N)and glucose monomers,the TPA-Glc N were efficiently assimilated by the bacterial cells and incorporated into the cellulose matrix,resulting in a uniform distribution of fluorescence.The fluorescence color and intensity of the obtained cellulose could be adjusted by varying the amount of the fluorescent glucose derivatives.Compared to the fluorescent cellulose synthesized through physical dyeing,the fluorescence of the products obtained by in situ polymerization showed higher intensity and stability.Furthermore,fluorescent bacterial cellulose can be hydrolyzed into nanocellulose-based ink,which demonstrates exceptional anti-counterfeiting capabilities under UV light.This biosynthesis method not only overcomes the limitations of traditional modification techniques but also highlights the potential of microbial systems as platforms for synthesizing functional polymers.展开更多
Methylglyoxal (MGO) is a vital signaling molecule that related to a variety of pathologies in both animals and plants. However, high levels of MGO are associated with several diseases. Therefore, developing a sensitiv...Methylglyoxal (MGO) is a vital signaling molecule that related to a variety of pathologies in both animals and plants. However, high levels of MGO are associated with several diseases. Therefore, developing a sensitive method for monitoring MGO levels in vivo and investigating its molecular mechanism is of great importance. Although most of the reported MGO fluorescence probes are designed for cells and animals, none have been used for study MGO levels in plants. Consequently, we herein report a fluorescent probe named CPDN, which is rational constructed utilizing coumarin derivatives and O-phenylenediamine as the fluorophore and the recognition group, respectively. In our study, CPDN have shown ability to selectively and sensitively detect MGO in solution and has been successfully exploited for imaging endogenous and exogenous MGO levels in living cells, zebrafish and Arabidopsis thaliana. Surprisingly, further investigation of CPDN has found that high MGO levels in Arabidopsis thaliana could inhibit the root growth. Moreover, it is demonstrated that the MGO levels in Arabidopsis thaliana increased when subjected to drought stress, which may be the main cause inhibiting root development and resulting in shorter root length. Therefore, the probe CPDN can be a powerful tool for studying the MGO levels under abiotic stress conditions and exploring its role in plant growth mechanisms. We believe that the application of CPDN in monitoring MGO levels in plants holds great values for deepening the understanding of plant growth mechanisms.展开更多
Organic small molecule fluorophores have been widely used in biology and biochemistry to study cellular structures and processes at high spatial and temporal resolution.Small-molecule dyes offer various benefits,such ...Organic small molecule fluorophores have been widely used in biology and biochemistry to study cellular structures and processes at high spatial and temporal resolution.Small-molecule dyes offer various benefits,such as high photostability,low molecular weight,and great biocompatibility.However,the poor brightness of most of conventional dyes in biological environments limits their use in high-quality superresolution fluorescence imaging.Chemists have conceived and developed many methods to enhance the brightness of fluorophores,including structural alterations that raise extinction coefficients and quantum yields.This review outlines current attempts and substantial advances achieved by chemists to improve the brightness of organic small-molecule fluorescent dyes,such as scaffold rigidification and twisted intramolecular charge transfer(TICT)inhibition.We think that this review will help researchers understand the chemical mechanisms involved in increasing the brightness of fluorophores for biological applications.展开更多
The fluorophores Xan-OH and Xan-OH/FBS,based on xanthene structure,possess an effective nearinfrared absorption,near-infraredⅡ(NIR-Ⅱ)fluorescent imaging ability,and excellent photothermal property.Xan-OH/FBS also ha...The fluorophores Xan-OH and Xan-OH/FBS,based on xanthene structure,possess an effective nearinfrared absorption,near-infraredⅡ(NIR-Ⅱ)fluorescent imaging ability,and excellent photothermal property.Xan-OH/FBS also has good viscosity-sensitivity,enabling the real-time in vivo visualization of acute liver injury induced by CCl_(4).Moreover,the photothermal conversion coefficient of Xan-OH and Xan-OH/FBS under 808 nm laser irradiation are significant(27.53%and 26.77%,respectively),which could realize NIR-Ⅱfluorescence imaging-guided photothermal therapy for He La xenograft tumor.Given these promising characteristics,Xan-OH/FBS is an efficient NIR-Ⅱfluorescent imaging agent for acute liver injury and a potential photothermal therapeutic agent for tumor.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
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
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 following grants:National Natural Science Foundation of China(grant nos.92354305,32271428,and 32201132)National Key R&D Program of China(grant no.2022YFC3401100)+1 种基金Fund for Knowledge Innovation of Wuhan Science and Technology Bureau(grant no.2022020801010558)Director Fund of WNLO.
文摘The rapid development of super-resolution microscopy has made it possible to observe subcellular structures and dynamic behaviors in living cells with nanoscale spatial resolution, greatly advancing progress in life sciences. As hardware technology continues to evolve, the availability of new fluorescent probes with superior performance is becoming increasingly important. In recent years, fluorescent nanoprobes (FNPs) have emerged as highly promising fluorescent probes for bioimaging due to their high brightness and excellent photostability. This paper focuses on the development and applications of FNPs as probes for live-cell super-resolution imaging. It provides an overview of different super-resolution methods, discusses the performance requirements for FNPs in these methods, and reviews the latest applications of FNPs in the super-resolution imaging of living cells. Finally, it addresses the challenges and future outlook in this field.
基金the financial support of this work by the National Natural Science Foundation of China(Nos.82072309,22067019 and 22367023)the Major Science and Technology Projects in Yunnan Province(No.202402AE090006)+3 种基金the Project of Yunnan Characteristic Plant Screening and R&D Service CXO Platform(No.2022YKZY001)Yunnan Provincial Science and Technology Department Yunnan University Joint Special Project(No.202201BF070001-001)the Postgraduate Research Innovation Foundation of Yunnan University(No.KC-23234403)the Scientific Research Foundation Project of Yunnan Provincial Department of Education(No.2023Y0240)。
文摘PBQ[1-(4-chlorophenyl)-3-(pyridin-3-yl)urea],an enormous potent molluscicide,showed excellent Pomacea canaliculata(P.canaliculata)control activity and low toxicity for other aquatic organisms,but its snail-killing mechanisms are still not fully understood.We employed an optical method to elucidate PBQ action via a novel fluorescent viscosity probe,NCV.As the viscosity in the test solutions increased,compared with that in pure ethanol,a 54-fold fluorescence intensity enhancement of NCV was observed in 310 cP of 90%glycerol.Furthermore,NCV successfully exhibited a selective fluorescence response towards monensin-induced cellular viscosity changes in HepG2 cells.The liver,stomach,and foot plantar of the tested snails were frozen and sectioned for fluorescent imaging experiments after the treatment with different PBQ concentrations over various times.A significant fluorescent increase in the snail's liver was observed upon exposure to 0.75 mg/L PBQ for 72 h,which highlighted an increase in viscosity.Hematoxylin and eosin(HE)staining further supported PBQ-induced liver damage with a viscosity increase in P.canaliculata.Our study provides a new rapid optical visualization method to study the killing mechanisms of PBQ and may help discover new chemicals that control snail populations.
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.
基金supported by the National Nature Science Foundation of China(Nos.62075079,62305127,61975200)the Natural Science Foundation of Jilin Province(20230508135RC)the Science and Technology Development Foundation of Changchun City(23GZZ15).
文摘The fluorescence imaging (FLI) in the second near-infrared window (NIR-II, 1000–1700nm) has attracted considerable attention in the past decade. In contrast to conventional NIR-I window excitation (808nm/980nm), FLI with NIR-II window excitation (1064nm/other wavelength beyond 1000nm) can afford deeper tissue penetration depth with high clarity due to the merits of suppressed photon scattering and diminished autofluorescence. In this review, we have summarized NIR-II window excitable/emissive organic/polymeric fluorophores recently developed. The characteristics of these fluorophores such as chemical structures and photophysical properties have also been critically discussed. Furthermore, the latest development of noninvasive in vivo FLI with NIR-II excitation was highlighted. The ideal imaging results emphasized the importance of NIR-II excitation of these fluorophores in enabling deep tissue penetration and high-resolution imaging. Finally, a perspective on the challenges and prospects of NIR-II excitable/emissive organic/polymeric fluorophores was also discussed. We expected this review will be served as a source of inspiration for researchers, stimulating the creation of novel NIR-II excitable fluorophores and fostering the development of bioimaging applications.
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We report five coordination polymers(CPs)based on fluorescent ligands[1,6-di(1H-imidazol-1-yl)pyrene(dip),9,10-di(1H-imidazol-1-yl)anthracene(dia)]and anionic ligands[cyclohexane-1,4-dicarboxylic acid(H_(2)cda),camphoric acid(H_(2)cpa)].In[Cd(dip)(cda)]·4H_(2)O}_(n)(1),the Cd^(2+)ions,acting as tetrahedral nodes,are linked by dipand cda^(2-)ligands with four Cd^(2+)ions into five-fold interpenetrating network array of topology of dia.In{[Cd(dip)(cpa)]·4H_(2)O}_(n)(2),the Cd^(2+)ions,acting as a 4-connector,are linked by cpa^(2-)and dip ligands into a 3D framework ofcds topology.In{[Ni(dia)_(2)Cl_(2)]·DMF}_(n)(3),the Ni^(2+)ion is linked by four dia ligands into a layer structure,and 1Dchannels of a cross-section of 1.35 nm×0.96 nm are formed.In{[Cd(dia)_(2)(H_(2)O)_(2)](NO_(3))_(2)·2DMSO}n(4),the dia ligandsconnected Cd^(2+)ions into a 2D layer,and 1D channels are formed between adjacent layers with a cross-section of0.87 nm×0.43 nm.In[Zn(dip)Cl_(2)]_(n)(5),the Zn^(2+)ion is linked by dip ligands into an infinite 1D chain.The infrared,thermal gravimetric,and fluorescent emission data were collected and analyzed for these coordination polymers.CCDC:2356055,1;2440075,2;2356057,3;2356057,4;2356059,5.
文摘Succinylcholine(SC)is a widely used depolarizing muscle relaxant,but improper use can lead to arrhythmias and,in severe cases,pose a life-threatening risk.Additionally,some criminals exploit SC for illicit activities.Therefore,rapid SC detection is paramount for clinical practice and public safety.Currently,however,limited methods are available for the rapid detection of SC.A fluorescent indicator displacement assay sensor based on molecular recognition of an amide naphthotube was developed.This sensor enabled the rapid fluorescent detection of SC through competitive binding between SC and methylene blue with the amide naphthotube.The sensor exhibited exceptional sensitivity with a detection limit as low as 1.1μmol/L and a detection range of 1.1~60μmol/L,coupled with outstanding selectivity and robust stability.Furthermore,this sensor accurately determined SC levels in biological samples such as serum.In summary,this research provides a new solution for the rapid and accurate sensing of SC in complex matrices and offers new insights for the swift identification and detection of toxins.
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Six coordination polymers based on 9,10-di(pyridine-4-yl)-anthracene(DPA)and 1,6-di(1H-imidazol-1-yl)pyrene(DIP)were obtained by solvothermal reactions.{[Zn(DPA)Cl_(2)]·DMF·2H_(2)O}n(1)and{[Zn_(1.5)(DPA)_(1.5)Cl_(3)]·5H_(2)O}n(2)are framework isomers,which both contain zigzag chains formed by DPA,Zn^(2+),and Cl-.The zigzag chains in 1 are further assembled by C—H…Cl interactions into layers,and these layers exhibit two different orientations,displaying a rare 2D to 3D interpenetration mode.The zigzag chains in 2 are parallelly arranged.{[Zn_(3)(DPA)_(3)Br_(6)]·2DMF·_(1.5)H_(2)O}n(3)is isostructural to 2.3 was obtained using ZnBr_(2)instead of ZnCl_(2).[M(DPA)(formate)_(2)(H_(2)O)_(2)]n[M=Co(4),Cu(5)]are isostructural,contain chain structures formed by DPA,Cu^(2+)/Co^(2+),and for-mate ions,which were formed in situ in the solvothermal reaction.{[Zn(DIP)_(2)Cl]ClO_(4)}n(6)contains a layer structure formed by DIP and Zn^(2+).Free DPA and DIP ligands exhibited high fluorescence at room temperature,and coordina-tion polymers 3 and 6 displayed enhanced fluorescent emissions.
基金the National Nature Science Foundation of China(No.22107028)Natural Science Foundation of Chongqing(No.CSTB2023NSCQ-MSX0335)。
文摘The rapid development of microfluidic technology has led to the evolution of microdroplets from simple emulsion structures to complex multilayered and multicompartmental configurations.These advancements have endowed microdroplets with the capability to contain multiple compartments that remain isolated from one another,enabling them to carry different molecules of interest.Consequently,researchers can now investigate intricate spatially confined chemical reactions and signal transduction pathways within subcellular organelles.Moreover,modern microdroplets often possess excellent optical transparency,allowing fluorescently labelled,multi-layered,and compartmental droplets to provide detailed insights through real-time,in situ,and dynamic fluorescence imaging.Hence,this review systematically summarizes current methodologies for preparing multicomponent microdroplets and their applications,particularly focusing on fluorescent microdroplets.Additionally,it discusses existing critical challenges and outlines future research directions.By offering a comprehensive overview of the preparation methods and applications of fluorescent microdroplets,this review aims to stimulate the interest of researchers and foster their utilization in more complex and biomimetic environments.
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 Natural Science Foundation of China(No.22376111)Shandong Provincial Natural Science Foundation(No.ZR2024YQ026)+2 种基金for Excellent Young Scholars,Taishan Scholar Foundation of Shandong Province(No.tsqn202408237)Youth Innovation Team Project for Talent Introduction and Cultivation in Universities of Shandong Province(No.096-1622002)the Research Foundation for Distinguished Scholars of Qingdao Agricultural University(No.663-1117015)。
文摘In recent years,cellulose-based fluorescent polymers have received considerable attention.However,conventional modification methods face challenges such as insolubility in most solvents,fluorescence instability,and environmental risks.In this study,a novel biosynthesis strategy was developed to fabricate fluorescent cellulose by adding fluorescent glucose derivatives to a bacterial fermentation broth.The metabolic activity of bacteria is utilized to achieve in situ polymerization of glucose and its derivatives during the synthesis of bacterial cellulose.Owing to the structural similarity between triphenylamine-modified glucose(TPA-Glc N)and glucose monomers,the TPA-Glc N were efficiently assimilated by the bacterial cells and incorporated into the cellulose matrix,resulting in a uniform distribution of fluorescence.The fluorescence color and intensity of the obtained cellulose could be adjusted by varying the amount of the fluorescent glucose derivatives.Compared to the fluorescent cellulose synthesized through physical dyeing,the fluorescence of the products obtained by in situ polymerization showed higher intensity and stability.Furthermore,fluorescent bacterial cellulose can be hydrolyzed into nanocellulose-based ink,which demonstrates exceptional anti-counterfeiting capabilities under UV light.This biosynthesis method not only overcomes the limitations of traditional modification techniques but also highlights the potential of microbial systems as platforms for synthesizing functional polymers.
基金support from the National Natural Science Foundation of China(No:21804102)the Open Project of Key Laboratory of Novel Biomass-Based Environmental and Energy Materials in Petroleum and Chemical Industry(No:BEEA0001)+1 种基金the Opening Project of Hubei Key Laboratory of Purification and Application of Plant Anti-cancer Active Ingredients(No:HLPAI2023011)the Graduate Innovative Fund of Wuhan Institute of Technology(No:CX2022429).
文摘Methylglyoxal (MGO) is a vital signaling molecule that related to a variety of pathologies in both animals and plants. However, high levels of MGO are associated with several diseases. Therefore, developing a sensitive method for monitoring MGO levels in vivo and investigating its molecular mechanism is of great importance. Although most of the reported MGO fluorescence probes are designed for cells and animals, none have been used for study MGO levels in plants. Consequently, we herein report a fluorescent probe named CPDN, which is rational constructed utilizing coumarin derivatives and O-phenylenediamine as the fluorophore and the recognition group, respectively. In our study, CPDN have shown ability to selectively and sensitively detect MGO in solution and has been successfully exploited for imaging endogenous and exogenous MGO levels in living cells, zebrafish and Arabidopsis thaliana. Surprisingly, further investigation of CPDN has found that high MGO levels in Arabidopsis thaliana could inhibit the root growth. Moreover, it is demonstrated that the MGO levels in Arabidopsis thaliana increased when subjected to drought stress, which may be the main cause inhibiting root development and resulting in shorter root length. Therefore, the probe CPDN can be a powerful tool for studying the MGO levels under abiotic stress conditions and exploring its role in plant growth mechanisms. We believe that the application of CPDN in monitoring MGO levels in plants holds great values for deepening the understanding of plant growth mechanisms.
基金supported by the National Science Foundation for Distinguished Young Scholars(No.22325401)the National Natural Science Foundation of China(No.22404049)the China Postdoctoral Science Foundation(No.2024M750866)。
文摘Organic small molecule fluorophores have been widely used in biology and biochemistry to study cellular structures and processes at high spatial and temporal resolution.Small-molecule dyes offer various benefits,such as high photostability,low molecular weight,and great biocompatibility.However,the poor brightness of most of conventional dyes in biological environments limits their use in high-quality superresolution fluorescence imaging.Chemists have conceived and developed many methods to enhance the brightness of fluorophores,including structural alterations that raise extinction coefficients and quantum yields.This review outlines current attempts and substantial advances achieved by chemists to improve the brightness of organic small-molecule fluorescent dyes,such as scaffold rigidification and twisted intramolecular charge transfer(TICT)inhibition.We think that this review will help researchers understand the chemical mechanisms involved in increasing the brightness of fluorophores for biological applications.
基金supported by the National Natural Science Foundation of China(Nos.22067021,22367024,and 22267023)Innovation Team of Yunnan Education Department+1 种基金the“Youth Talent of Wan Ren Project”the Sichuan Science and Technology Program(No.2022YFS0633)。
文摘The fluorophores Xan-OH and Xan-OH/FBS,based on xanthene structure,possess an effective nearinfrared absorption,near-infraredⅡ(NIR-Ⅱ)fluorescent imaging ability,and excellent photothermal property.Xan-OH/FBS also has good viscosity-sensitivity,enabling the real-time in vivo visualization of acute liver injury induced by CCl_(4).Moreover,the photothermal conversion coefficient of Xan-OH and Xan-OH/FBS under 808 nm laser irradiation are significant(27.53%and 26.77%,respectively),which could realize NIR-Ⅱfluorescence imaging-guided photothermal therapy for He La xenograft tumor.Given these promising characteristics,Xan-OH/FBS is an efficient NIR-Ⅱfluorescent imaging agent for acute liver injury and a potential photothermal therapeutic agent for tumor.
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 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.