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Simple imaging for the diamond anvil cell: Applications to hard-to-reach places
摘要： The employment of high-pressure gases as a pressure-transmitting medium, sample, or reactant for diamond anvil cell experiments is widespread. As a pressure transmitter, high-pressure gases are crucial to forming quasi-hydrostatic compression atmospheres for samples inside the uniaxially driven cell. We describe an optical design for forming high-resolution images of the gasket and sample chamber of the diamond anvil cell under high gas pressures in a gas loading apparatus. Our design is simple, is of low-cost, and may be easily adapted to suit gas loading apparatus of any design, as well as other common hard-to-reach environments in diamond anvil cell experiments, i.e., those with large stand-off distances, such as cryostats.
关键词： optical design,high-pressure gases,gas loading apparatus,diamond anvil cell,imaging
更新于2025-09-23 15:21:21
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Selective and Sensitive Sensing of Hydrogen Peroxide by a Boronic Acid Functionalized Metal–Organic Framework and Its Application in Live-Cell Imaging
摘要： A new boronic acid functionalized Zr(IV) metal?organic framework having the capability of sensing H2O2 in live cells is reported. The Zr-MOF bears a UiO-66 structure and contains 2-boronobenzene-1,4-dicarboxylic acid (BDC-B(OH)2) as a framework linker. The activated Zr-UiO-66-B(OH)2 compound (called 1′) is highly selective for the ?uorogenic detection of H2O2 in HEPES bu?er at pH 7.4, even in the presence of interfering ROS (ROS = reactive oxygen species) and other biologically relevant analytes. The ?uorescent probe was found to display extraordinary sensitivity for H2O2 (detection limit 0.015 μM) in HEPES bu?er, which represents a lower value in comparison to those of the MOF probes documented so far for sensing H2O2 using other analytical methods. Taking advantage of its high selectivity and sensitivity for H2O2 in HEPES bu?er, the probe was successfully employed for the imaging of intracellular H2O2. Imaging studies with MDAMB-231 cells revealed the emergence of bright blue ?uorescence after loading with probe 1′ and subsequent treatment with H2O2 solution.
关键词： Zr(IV) metal?organic framework,boronic acid,live-cell imaging,fluorescent probe,H2O2 sensing
更新于2025-09-23 15:21:21
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Aprotinin Encapsulated Gold Nanoclusters: A Fluorescent Bioprobe with Dynamically Nuclear Targeting and Selective Detection of Trypsin and Heavy Metal
摘要： Fluorescence imaging has currently emerged as one of the most frequently used noninvasive imaging technology to selectively monitor biological processes in living systems. In past decades, gold nanoclusters (Au NCs) received increasing attraction because of their intrinsic fluorescence and their inherent biocompatibility. As a stabilizing and reducing agent, an abundant, sustainable and widely used polypeptide derived drug molecule, aprotinin (Ap) is selected for the synthesis of Au nanoclusters (Ap-Au NCs) due to characteristic bioactivity, excellent biocompatibility, biodegradability, nonallergenic character. Herein, Ap encapsulated Au NCs exhibiting desirable red fluorescence feature was facilely produced for the first time, which were subsequently used for cell imaging and detection of various analytes. Much interestingly, dynamically subcellular localization of Ap-Au NCs from the cytoplasm to the nucleus in Hela cells was observed. Afterward, it has shown the selective and quantitative detection of trypsin by using Ap encapsulated Au NCs. Lastly, Ap-Au NCs were readily used for detection of mercury and copper quantitatively. The photoluminescence of the Ap-Au NCs was quenched with the addition of metioned analytes. This study opens crucial insights on the integration of biomolecule with metal nanoclusters, also discusses a multifunctional nanomaterial platform for cell imaging, subcellular targeting, biosensing and drug delivery.
关键词： trypsin detection,gold nanoclusters,aprotinin,cell imaging,heavy metal detection,Fluorescence imaging
更新于2025-09-23 15:21:01
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Quaternary Supramolecular Nanoparticles as a Photoerasable Luminescent Ink and Photocontrolled Cella??Imaging Agent
摘要： Photoswitchable luminescent supramolecular assemblies based on cyclodextrins have attracted considerable attention owing to their potential applications as smart materials, but most of the assemblies reported to date emit green or blue light with low contrast and high interference. In this study, novel photoluminescent red-luminescent quaternary supramolecular nanoparticles (2) are constructed from a dithienylethene derivative (1), a β-cyclodextrin-functionalized ruthenium complex (Ru-HOP-CD), Pluronic F-127, and cetrimonium bromide. Compared with the binary assembly 1@Ru-HOP-CD, the quaternary nanoparticles exhibit high fluorescence resonance energy transfer efficiency, with Ru-HOP-CD acting as the donor and 1 as the acceptor. Owing to the reversible photoswitched interconversion of the two forms of the dithienylethene component, the fluorescence of the nanoparticles could be switched on/off by irradiation with UV or visible light, both in solution and in the solid state. As a result, the nanoparticles could be used as a photoerasable red-luminescent ink and as a photocontrolled cell-imaging agent. These functional nanoparticles can be expected to be useful in the fields of information security and biology.
关键词： anticounterfeiting,cell imaging,cyclodextrin,photoerasable luminescent ink,diarylethene
更新于2025-09-23 15:21:01
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Development of a Selective Fluorescent Probe for Hypochlorous Acid Detection and Imaging
摘要： The reactive oxygen species (ROS), hypochlorous acid (HOCl), participates in many important biological events. For example, this ROS is the main active agent to kill invading microbes by immune system. HOCl is produced by H2O2 and Cl? in a myeloperoxidase (MPO) catalyzed process that occurs in response to inflammatory stimuli. Because HOCl reacts with important biological molecules and it causes cell damage, its uncontrolled production is involved in some human maladies such as arthritis. Detection and imaging HOCl in vivo are critical components of studies aimed at elucidating the biological functions of this ROS. Owing to several advantageous features including high selectivities and sensitivities, short response times and simple preparation methods, molecular fluorescent probes have become powerful tools for detecting biologically important substances and bio-imaging. It is essential to develop highly selective and sensitive fluorescent probes for HOCl for bio-applications. In the study described below, we demonstrated that spiro[isobenzofuran-30,60-dihydroxy-1(3H),90-[9H]thioxanthen]-3-one (TF) is an ideal fluorescent probe for HOCl because it undergoes a fluorescence enhancement upon being selectively chlorinated by this ROS.
关键词： Cell imaging,Fluorescent probe,Hypochlorous acid detection
更新于2025-09-23 15:21:01
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In situ imaging of live-cell extracellular pH during cell apoptosis with SERS
摘要： Extracellular pH (pHe) is an important regulating factor that determines many cellular processes, including proliferation, differentiation, and apoptosis. In our previous work, we developed 4-MPy (4-mercaptopyridine) modified Au nanoparticles as intracellular pH sensors based on surface-enhanced Raman spectroscopy (SERS). We herein modified Au nanoparticles-assembled solid SERS substrate with 4-MPy molecules for in situ pHe sensing during apoptosis. We found a more acidic extracellular environment of cancer cells than that of normal cells from the pH imaging. We then in situ investigated the temporal and spatial evolution of pHe of cancer cells after addition of transforming growth factor-β (TGF-β). The pHe showed a fast decrease at the beginning, followed by a slow decrease until the complete loss of cellular functions and the pH values in and out of the cells became similar. This work shows that our SERS substrate combined with in situ cell culture system is well suitable for in situ pHe sensing during cell processes and will be a promising technique for understanding more pHe-related biological and pathological issues.
关键词： apoptosis,live-cell imaging,TGF-β,extracellular pH,SERS
更新于2025-09-23 15:21:01
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Fast response fluorogenic probe for visualizing hypochlorite in living cells and Zebrafishes
摘要： A novel fast response and visible fluorogenic probe (D1) for monitoring hypochlorite (ClO?) has been developed based on specific ClO? removal of a C=N bond. The response of the probe to ClO? showed increases linearly, and the fluorescence intensity heighten about 25 times. D1 responses to ClO? were observed with high selectivity and sensitivity within 10 s by naked eyes. D1 can not only detect the level of hypochlorite in vitro such as urine, but also monitoring hypochlorite content in extremely cold condition as low as -78oC. Meanwhile, the good biocompatibility guaranteed the use of D1 to detect intracellular ClO? under confocal microscopy. Moreover, D1 has been successfully applied to monitor the exogenous and endogenous ClO? in Zebrafishes with fluorescence imaging.
关键词： Zebrafishes,live cell imaging,hypochlorite,fast response,fluorogenic probe
更新于2025-09-23 15:21:01
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Design of Magnetica??Plasmonic Nanoparticle Assemblies via Interface Engineering of Plasmonic Shells for Targeted Cancer Cell Imaging and Separation
摘要： Magnetic-plasmonic nanoparticles have received considerable attention for widespread applications. These nanoparticles (NPs) exhibiting surface-enhanced Raman scattering (SERS) activities are developed due to their potential in bio-sensing applicable in non-destructive and sensitive analysis with target-specific separation. However, it is challenging to synthesize these NPs that simultaneously exhibit low remanence, maximized magnetic content, plasmonic coverage with abundant hotspots, and structural uniformity. Here, a method that involves the conjugation of a magnetic template with gold seeds via chemical binding and seed-mediated growth is proposed, with the objective of obtaining plasmonic nanostructures with abundant hotspots on a magnetic template. To obtain a clean surface for directly functionalizing ligands and enhancing the Raman intensity, an additional growth step of gold (Au) and/or silver (Ag) atoms is proposed after modifying the Raman molecules on the as-prepared magnetic-plasmonic nanoparticles. Importantly, one-sided silver growth occurred in an environment where gold facets are blocked by Raman molecules; otherwise, the gold growth is layer-by-layer. Moreover, simultaneous reduction by gold and silver ions allowed for the formation of a uniform bimetallic layer. The enhancement factor of the nanoparticles with a bimetallic layer is approximately 107. The SERS probes functionalized cyclic peptides are employed for targeted cancer-cell imaging and separation.
关键词： cell separation,surface-enhanced Raman scattering,bio-sensing,magnetic-plasmonic nanoparticles,cancer-cell imaging
更新于2025-09-23 15:19:57
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Surface plasmona??coupled emission imaging for biological applications
摘要： Fluorescence imaging technology has been extensively applied in chemical and biological research profiting from its high sensitivity and specificity. Much attention has been devoted to breaking the light diffraction–limited spatial resolution. However, it remains a great challenge to improve the axial resolution in a way that is accessible in general laboratories. Surface plasmon–coupled emission (SPCE), generated by the interactions between surface plasmons and excited fluorophores in close vicinity of the thin metal film, offers an opportunity for optical imaging with potential application in analysis of molecular and biological systems. Benefiting from the highly directional and distance-dependent properties, SPCE imaging (SPCEi) has displayed excellent performance in bioimaging with improved sensitivity and axial confinement. Herein, we give a brief overview of the development of SPCEi. We describe the unique optical characteristics and constructions of SPCEi systems and highlight recent advances in the use of SPCEi for biological applications. We hope this review provides readers with both the insights and future prospects of SPCEi as a new promising imaging platform for potentially widespread applications in biological research and medical diagnostics.
关键词： Fluorescence microscopy,Directional emission,Background suppression,Surface plasmon–coupled emission imaging,Biological application,Cell imaging
更新于2025-09-23 15:19:57
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A waveguide imaging platform for livea??cell TIRF imaging of neurons over large fields of view
摘要： Large fields of view (FOVs) in total internal reflection fluorescence microscopy (TIRFM) via waveguides have been shown to be highly beneficial for single molecule localisation microscopy on fixed cells [1, 2] and have also been demonstrated for short-term live-imaging of robust cell types [3-5], but not yet for delicate primary neurons nor over extended periods of time. Here, we present a waveguide-based TIRFM set-up for live-cell imaging of demanding samples. Using the developed microscope, referred to as the ChipScope, we demonstrate successful culturing and imaging of fibroblasts, primary rat hippocampal neurons and axons of Xenopus retinal ganglion cells (RGCs). The high contrast and gentle illumination mode provided by TIRFM coupled with the exceptionally large excitation areas and superior illumination homogeneity offered by photonic waveguides have potential for a wide application span in neuroscience applications.
关键词： live-cell imaging,TIRFM,neurons,large FOV,waveguide
更新于2025-09-23 15:19:57