We propose a direct electron-beam excitation assisted optical microscope with resolution of few tens nanometers and it can be applied for observation dynamic movements nanoparticles in liquid. The technique is also useful live cell imaging under physiological conditions as well colloidal solution, microcrystal growth solutions, etc. In the microscope, fluorescent materials are directly excited focused electron beam. an beam yields high spatial since to specimens. order demonstrate potential...

Although investigating drug modulation of cytochrome P450 (CYP) activity under physiological conditions is crucial in development to avoid severe adverse reactions, the current evaluation approaches that rely on destructive and end-point analysis can be misleading due invasive treatments cellular heterogeneity. Here, we propose a non-destructive high-content method for visualizing quantifying intracellular CYP administration by Raman microscopy. The redox-state spin-state sensitive...

Abstract Multi‐color, high spatial resolution imaging of fluorescent nanodiamonds (FNDs) in living HeLa cells has been performed with a direct electron‐beam excitation‐assisted fluorescence (D‐EXA) microscope. In this technique, materials are directly excited focused electron beam and the resulting cathodoluminescence (CL) is detected nanoscale resolution. Green‐ red‐light‐emitting FNDs were employed for two‐color imaging, which observed simultaneously This technique could be applied...

We present a technique for improving the spatial resolution of two-photon excitation microscopy; our combines annular illumination with an in situ estimation point spread function (PSF) used deconvolution. For PSF, we developed called autocorrelation scanning, which sample is imaged by scanning two foci that are overlapped over various distances. The image series obtained variation distance between provides can be to estimate PSF at specific positions within sample. proved principle and...

Simultaneous observation of drug distribution at the effector site and subsequent cell response are essential in development process. However, few studies have visualized itself biomolecular interactions living cells. Here, we used label-free Raman microscopy to investigate drug-induced cytotoxicity visualize uptake subcellular localization by its specific molecular fingerprint. A redox-sensitive microscope detected decrease reduced cytochrome c (cyt c) after Actinomycin D (ActD) treatment a...

The transmission of neuronal information is propagated through synapses by neurotransmitters released from presynapses to postsynapses. Neurotransmitters the presynaptic vesicles activate receptors on postsynaptic membrane. Glutamate acts as a major excitatory neurotransmitter for synaptic in central nervous system. Determining concentration glutamate single essential understanding mechanisms activation normal brain functions well neurological diseases. However, it difficult detect and...

We demonstrated resolution improvement in two-photon excitation microscopy by combining saturated (SAX) of fluorescence and pupil manipulation. theoretically estimated the sidelobe effect point spread function with various designs found that combination SAX core-ring illumination can effectively enhance spatial 3D suppress artifacts. The experimental demonstration shows proposed technique is effective for observation a depth 100 μm tissue phantom be applied to observations samples higher...

We report a method to increase the efficiency of detecting nonlinear fluorescence signals in saturated excitation (SAX) microscopy. With this method, we compare obtained under different degrees extract fluorescent signal induced by excitation. Compared conventional SAX microscopy using harmonic demodulation technique, detection can be increased up 8 and 32 times imaging second-order third-order signals, respectively. combined approach with pulsed excitation, which is effective reduce...

To promote the clinical application of human induced pluripotent stem cell (hiPSC)-derived hepatocytes, a method capable monitoring regenerative processes and assessing differentiation efficiency without harming or modifying these cells is important. Raman microscopy provides powerful tool for this as it enables label-free identification intracellular biomolecules in live samples. Here, we used to assess hiPSC into hepatocyte lineage based on chemical content. We contrasted data with similar...

We developed a high-resolution fluorescence microscope in which fluorescent materials are directly excited using focused electron beam. Electron beam excitation enables detailed observations on the nanometer scale. Real-time live-cell observation is also possible thin film to separate environment under study from vacuum region required for propagation. In this study, we demonstrated of cellular components by autofluorescence with and performed dynamic intracellular granules. Since associated...

We fabricated ZnO/SiN films for use as a light source of high-resolution optical microscope and characterized the properties films, demonstrated images obtained with using films. A 100-nm-thick ZnO film deposited on SiN showed much higher CL intensity than film, it was enhanced by high-temperature annealing film. Electron beam excitation assisted gold particles 200 nm diameter taken indicated that can provide signal-to-noise (S/N) ratio frame rate It shown dynamic observation living cells...

Abstract Optical microscopes are effective tools for cellular function analysis because biological cells can be observed non-destructively and non-invasively in the living state either water or atmosphere condition. Label-free optical imaging technique such as phase-contrast microscopy has been analysed many functions it is essential technology bioscience field. However, diffraction limit of light makes difficult to image nano-structures a label-free cell, example endoplasmic reticulum,...

High spatial resolution microscope is desired for deep understanding of cellular functions, in order to develop medical technologies. We demonstrate high-resolution imaging un-labelled organelles living cells, which live cells on a 50 nm thick silicon nitride membrane are imaged by autofluorescence excited with focused electron beam through the membrane. Electron excitation enables ultrahigh organelles, such as mitochondria, nuclei, and various granules. Since spectra represent molecular...

Label‐free optical nano‐imaging of dendritic structures and intracellular granules in biological cells is demonstrated using a bright homogeneous nanometric light source. The source excited focused electron beam. A zinc oxide (ZnO) luminescent thin film was fabricated by atomic layer deposition (ALD) to produce the nanoscale ZnO formed ALD emitted bright, light, unlike that deposited another method. label‐free macrophage receptor with collagenous structure‐expressing CHO were clearly...

The surface plasmon resonance (SPR) technique has been widely applied to biosensing technologies for the rapid quantification of biomolecules without enzyme and fluorescent labeling. However, conventional prism-coupling SPR method generally a detection area few mm2, large contribution background signal forms barrier highly sensitive detection. Based on spatially resolved method, present study constructed scanning GC-SPR imaging instrument using an objective lens with high numerical aperture...

We fabricated a bright and thin Zn₂SiO₄ luminescent film to serve as nanometric light source for high-spatial-resolution optical microscopy based on electron beam excitation. The was by annealing ZnO Si₃N₄ substrate at 1000 °C in N₂. annealed emitted cathodoluminescence compared with the as-deposited film. is promising nano-imaging excitation-assisted microscopy. evaluated spatial resolution of microscope developed using this This first report investigation application ZnO/Si₃N₄ high...

We present an Electron-beam-eXcitation-Assisted (EXA) optical microscope with a nanometric illumination light source consisting of red cathode luminescence (CL) lights emitted by Y2O3:Eu3+ phosphor thin film excited high-energy focused electron beams. Phosphor films few hundred nanometers thick were fabricated on 50-nm Si3N4 membranes using beam evaporation. The preparation conditions for brighter CL emissions examined in terms the post-annealing temperatures and thickness. succeeded...

The excitatory synaptic transmission is mediated by glutamate (GLU) in neuronal networks of the mammalian brain. In addition to GLU, extra-synaptic GLU known modulate activity. networks, uptake an important role neurons and glial cells for lowering concentration extracellular avoid excitotoxicity. Monitoring spatial distribution intracellular study but approach has been hampered absence appropriate analogs that report localization GLU. Deuterium-labeled (GLU-D) a promising tracer monitoring...

The light that is illuminated on a silver-film-coating substrate with periodic structure, i.e., plasmonic chip, can couple to plasmon polaritons and enhance the electric field surface of chip. Fluorescent molecules fixed pattern are excited by an enhanced field, enhancing their fluorescence. Particularly, bright fluorescence point appears at center concentric circle called Bull's eye pattern. This nanoantenna effect has been studied in various types circles comprehended constructive wave...

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Developing a highly sensitive and accurate method to discriminate between amino acids peptides is vital for establishing future healthcare testing technologies, such as liquid biopsy. This study proposes technique based on surface-enhanced Raman scattering (SERS), which combines chemically linking an analyte with gold nanoparticles aggregating them produce hotspots. Furthermore, by combining rapid SERS imaging slit-scanning microscopy deep learning convolutional neural network, 20...

Abstract Plasmonic chips are chip-type devices with wavelength-order periodic structures covered metal based on grating-coupled surface plasmon resonance for sensitive detection. In this study, we investigated a pillar-array-type (PA) structure, which is an inversion of the hole-array-type (HA) to further improve fluorescence enhancement. Increases in dip depth reflection spectra and corresponding field enhancement were observed PA resulting larger than that HA structure. The relationship...

Intracellular structures of HeLa cells are observed using a direct electron beam excitation-assisted fluorescence (D-EXA) microscope.In this microscope, silicon nitride membrane is used as culture plate, which typically has low biocompatibility between the sample and surface to prevent from adhering strongly surface.In work, modified allow strong cell attachment, enables high-resolution observation intracellular an increased signal-to-noise ratio.In addition, penetration depth evaluated...