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A new lysosome-targetable fluorescent probe with a large Stokes shift for detection of endogenous hydrogen polysulfides in living cells
摘要： Hydrogen polysulfides (H2Sn, n>1) influence a variety of important biological functions and activities associated with hydrogen sulfide (H2S). The development of probes for rapid, selective, and sensitive detection of H2Sn still remains a great challenge. Lysosome plays crucial roles in various physiological processes among living cells, which arouse high interest in detecting endogenous lysosome-targetable H2Sn. To the best of our knowledge, there is no lysosome-targetable probe for H2Sn has been reported. In this work, a new lysosome-targetable probe NIPY-NF, based on the scaffold of imidazo[1,5-α]pyridine, with a large Stokes shift (215 nm), low detection limit (84 nM), fast response time (6 min) and low cytotoxicity was designed and synthesized. Furthermore, NIPY-NF was successfully applied into the cell imaging for detection of endogenous lysosome-targetable H2Sn.
关键词： Hydrogen polysulfides,Fluorescent probe,Lysosome,Cell imaging,Imidazo[1,5-α]pyridine
更新于2025-09-23 15:23:52
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A colorimetric and fluorometric oligothiophene-indenedione-based sensor for rapid and highly sensitive detection of cyanide in real samples and bioimaging in living cells
摘要： A new simple oligothiophene-indenedione-based sensor 3TI has been synthesized for the highly reactive and selective detection of cyanide anion (CN?) in 70% aqueous media. The sensor 3TI displays distinct colorimetric and fluorometric detection of CN? due to the addition of CN? to the electron-deficient indenedione-vinyl group of 3TI, which hampers intramolecular charge transfer (ICT) efficiencies. The recognition mechanism of 3TI for CN? was confirmed by the optical measurements, 1H NMR titration, FTIR spectra, HRMS analysis, and TD-DFT calculations. The sensor 3TI for CN? shows the outstanding advantages of high fluorescence brightness, fast response time (30 s), low detection limit (31.3 nM), minimal pH dependence in the physiologically relevant range, and excellent selectivity in presence of other competitive anions. Encouraged by these desirable sensing properties, the 3TI has been successfully used for determination of CN? in real water and food samples, silica-based sensing kits and fluorescence imaging in living cells with satisfactory results.
关键词： Fluorometric sensor,Cyanide,Oligothiophene-indenedione,Colorimetric sensor,Real sample,Live-cell imaging
更新于2025-09-23 15:23:52
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A genetically encoded FRET biosensor for visualizing EphA4 activity in different compartments of the plasma membrane
摘要： The EphA4 receptor tyrosine kinase is well known for its pivotal role in development, cancer progression, and neurological disorders. However, how EphA4 kinase activity is regulated in time and space still remains unclear. To visualize EphA4 activity in different membrane microdomains, we developed a sensitive EphA4 biosensor based on F?rster resonance energy transfer (FRET), and targeted it in or outside raft-like microdomains in the plasma membrane. We showed that our biosensor can produce a robust and specific FRET response upon EphA4 activation, both in vitro and in live cells. Interestingly, we observed stronger FRET responses for the non-raft targeting biosensor than for the raft targeting biosensor, suggesting that stronger EphA4 activation may occur in non-raft regions. Further investigations revealed the importance of the actin cytoskeleton in suppressing EphA4 activity in raft-like microdomains. Therefore, our FRET-based EphA4 biosensor could serve as a powerful tool to visualize and investigate EphA4 activation and signaling in specific subcellular compartments of single live cells.
关键词： EphA4,Membrane microdomain,cytoskeleton,FRET biosensor,Live cell imaging
更新于2025-09-23 15:22:29
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European Microscopy Congress 2016: Proceedings || Germ-line cysts organisation in clitellate annelids - from electron microscopy to live cell imaging
摘要： (cid:42)(cid:72)(cid:85)(cid:80)(cid:16)(cid:79)(cid:76)(cid:81)(cid:72)(cid:3)(cid:70)(cid:92)(cid:86)(cid:87)(cid:86)(cid:3)(cid:82)(cid:85)(cid:74)(cid:68)(cid:81)(cid:76)(cid:86)(cid:68)(cid:87)(cid:76)(cid:82)(cid:81)(cid:3)(cid:76)(cid:81)(cid:3)(cid:70)(cid:79)(cid:76)(cid:87)(cid:72)(cid:79)(cid:79)(cid:68)(cid:87)(cid:72)(cid:3)(cid:68)(cid:81)(cid:81)(cid:72)(cid:79)(cid:76)(cid:71)(cid:86)(cid:3)(cid:98)(cid:3)(cid:73)(cid:85)(cid:82)(cid:80)(cid:3)(cid:72)(cid:79)(cid:72)(cid:70)(cid:87)(cid:85)(cid:82)(cid:81)(cid:80)(cid:76)(cid:70)(cid:85)(cid:82)(cid:86)(cid:70)(cid:82)(cid:83)(cid:92)(cid:3)(cid:87)(cid:82)(cid:3)(cid:79)(cid:76)(cid:89)(cid:72)(cid:3)(cid:70)(cid:72)(cid:79)(cid:79)(cid:3)(cid:76)(cid:80)(cid:68)(cid:74)(cid:76)(cid:81)(cid:74)
关键词： live cell imaging,microcopy,germ-line cells
更新于2025-09-23 15:22:29
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A selective diaminomaleonitrile-based dual channel emissive probe for Al3+ and its application in living cell imaging
摘要： A selective diaminomaleonitrile-based fluorescent probe for Al3+(L) has been synthesized and characterized. The sensing behavior of L towards different metal ions was investigated by fluorescent spectra and competition experiment. The results showed that L exhibited a dual channel emission (λem 522 & 558 nm) with yellow fluorescence in the presence of Al3+ over other metal ions in EtOH solution. The probe L was bound to Al3+ in a 1: 2 stoichiometric manner as supported by Job’s plot analysis. The binding constant was 9.55×108 M-2 with the detection limit of 10.3 nM. The turn-on response was attributed to the inhibition of PET and C=N isomerization after binding with Al3+ while the dual emissions were due to ESIPT and ICT processes. In addition, the application of L for imaging Al3+ in living Hela cells was achieved.
关键词： diaminomaleonitrile-based,Al3+,living cell imaging,selective,dual channel
更新于2025-09-23 15:22:29
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An ESIPT-based fluorescent probe for Hg2+ in aqueous solution and its application in live-cell imaging
摘要： A new Excited-State Intramolecular Proton Transfer (ESIPT) based fluorescent probe for the detection of Hg2+ has been rationally designed and developed. Based on the specific reactivity of mercury-promoted hydrolysis, the probe exhibits high selectivity and sensitivity for mercury ions in almost pure aqueous solution (containing only 1% DMSO) with a low detection limit of 1.9 ppb. Furthermore, the probe was also successfully used for fluorescence imaging of Hg2+ in live cells.
关键词： Fluorescent probe,ESIPT-based,Live-cell imaging,Mercury ions,3-Hydroxyphthalimide
更新于2025-09-23 15:22:29
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A Strategy to Optimize the Generation of Stable Chromobody Cell Lines for Visualization and Quantification of Endogenous Proteins in Living Cells
摘要： Single-domain antibodies have emerged as highly versatile nanoprobes for advanced cellular imaging. For real-time visualization of endogenous antigens, fluorescently labelled nanobodies (chromobodies, CBs) are introduced as DNA-encoded expression constructs in living cells. Commonly, CB expression is driven from strong, constitutively active promoters. However, high expression levels are sometimes accompanied by misfolding and aggregation of those intracellular nanoprobes. Moreover, stable cell lines derived from random genomic insertion of CB-encoding transgenes bear the risk of disturbed cellular processes and inhomogeneous CB signal intensities due to gene positioning effects and epigenetic silencing. In this study we propose a strategy to generate optimized CB expressing cell lines. We demonstrate that expression as ubiquitin fusion increases the fraction of intracellularly functional CBs and identified the elongation factor 1α (EF1-α) promoter as highly suited for constitutive CB expression upon long-term cell line cultivation. Finally, we applied a CRISPR/Cas9-based gene editing approach for targeted insertion of CB expression constructs into the adeno-associated virus integration site 1 (AAVS1) safe harbour locus of human cells. Our results indicate that this combinatorial approach facilitates the generation of fully functional and stable CB cell lines for quantitative live-cell imaging of endogenous antigens.
关键词： chromobodies,live-cell imaging,cellular models,compound screening,nanobodies
更新于2025-09-23 15:22:29
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Two novel rhodamine-based fluorescent probes for the rapid and sensitive detection of Fe3+: Experimental and DFT calculations
摘要： Fe3+ ions play an important role in both biological and environmental field. In this work, two novel rhodamine-based colorimetric and fluorescent probes (RBA2 and RBA3) were designed and synthesized for the efficient detection of Fe3+. Upon the addition of Fe3+, the fluorescence intensity of RBA2 and RBA3 enhanced 108-fold and 222-fold, respectively. RBA2 and RBA3 exhibited a low detection limit which could achieve 12.8 nM and 11.0 nM. In addition, the binding modes of RBA2 and RBA3 with Fe3+ were proved to be 1:1 stoichiometry in the complexes by Job’s plot, ESI-MS and 1H NMR results. The complexing ability of RBA3 with Fe3+ excessed to that of RBA2 that was determined by the binding association constants, and highly consistent with DFT calculations results. Furthermore, RBA2 and RBA3 were further utilized to detect Fe3+ in living cells and real water samples, indicating their promising prospects in biological and environmental field.
关键词： Cell imaging,DFT calculation,Binding ability,Naked eye,Fe3+
更新于2025-09-23 15:22:29
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A novel ratiometric and colorimetric fluorescent probe for hydrazine based on ring-opening reaction and its applications
摘要： In recent years, with the increasing awareness of environmental protection, the detection of industrial emissions has become more and more strict. Hydrazine, as a common reactive group, the excessive emissions by the widespread use is inevitably harmful to the environment and human beings. To respond positively to environment protection policies at home and abroad, the development of a convenient and effective hydrazine probe has become an important subject for many research groups. In this work, a derivative containing lactone ring was synthesized based on naphthalimide. This probe displayed a fast colorimetric and ratiometric fluorescent response for hydrazine with high selectivity and sensitivity based on the ring-opening reaction. The detection limit of the probe for hydrazine is about 0.2032 μM (< 10 ppb). It is worth mentioning that the probe can successfully apply to detect gaseous hydrazine when the probe is loaded on the periopaper. Furthermore, the results of biological imaging experiment indicated that the probe can detect the hydrazine at the cellular level.
关键词： Cell imaging,Lactone ring,Hydrazine,Ring-opening,Ratiometric and colorimetric
更新于2025-09-23 15:22:29
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A rhodamine-deoxylactam based fluorescent probe for fast and selective detection of nitric oxide in living cells
摘要： Nitric oxide (NO) plays vital roles in many physiological process and is closely related to many diseases. So far, a number of fluorescent probes have been constructed for the detection of NO successfully. However, the probes still suffer from long-time reaction and limited selectivity. Herein, a fluorescent probe named dRB-OPD is synthesized and used to recognize NO. The probe contains a deoxy-rhodamine B as fluorophore and o-phenylenediamino as reaction site. dRB-OPD shows fast response to NO within 40 seconds with 170-fold fluorescence enhancement. Moreover, the probe shows high selectivity towards NO over dehydroascorbic acid (DHA), ascorbic acid (AA), and methylglyoxal (MGO). Particularly, the probe can avoid the serious interference from cysteine (Cys) found in the rhodamine lactam-based fluorescent NO probes (RB-OPD). In addition, the probe is applied for the detection of exogenous and endogenous NO in the HepG2 and RAW 264.7 cells with satisfactory results.
关键词： Cell imaging,Fluorescent probe,Nitric oxide,High selectivity,Fast response
更新于2025-09-23 15:22:29