Comparison of the higher diffraction order images in methyl-red-doped liquid crystal films

We investigate the change of higher diffraction order images in holographic image storage and reconstruction process. In experiments, an s-polarized Ar⁺ laser (488.0 nm) was used to record permanent grating in the dye-doped liquid crystal, 4,4′-n-entylcyanobiphenyl (5CB) doped with 1 wt% methyl-red (MR), at a small incident angle. Higher diffraction order images were observed when the signal beam was focused in front of and behind the film. Then the film was illuminated by an s-polarized He–Ne laser (632.8 nm) in the direction perpendicular to the surface. The higher diffraction order images were reconstructed. A theory about the change of higher diffraction order images is developed, which is in good agreement with experimental results. The results show that the higher diffraction order provides a useful method for optical information processing.     

Ultrasound-enhanced chemiluminescence tomography in biological tissue

This paper reports ultrasound-assisted optical imaging of chemiluminescent probes in biological tissue. A focused low power ultrasound sonochemically enhances a peroxyoxalate chemiluminescence (CL) that involves indocyanine green (ICG) as luminescent pigments. By scanning the focus, it produces tomographic images of CL in scattering media. The authors demonstrate imaging using a slab of porcine muscle measuring 50 × 50 × 75 mm, in which a capsuled CL reagent is embedded at 25 mm depth. Spatial resolution of imaging and concentration characteristics of CL reagents to enhanced CL intensity are also studied to evaluate the potential for use in bio-imaging applications with exploring the CL enhancement mechanisms. CL enhancement ratio, defined as the ratio of ultrasonically enhanced CL intensity to the base intensity without ultrasound irradiation, was found to be constant even in varying ICG and oxidizer concentrations, implying to be applicable for quantitative determination of these molecules.     

Image quality of microns-thick specimens in the ultra-high voltage electron microscope

Image quality of MeV transmission electrons is an important factor for both observation and electron tomography of microns-thick specimens with the high voltage electron microscope (HVEM) and the ultra-HVEM. In this work, we have investigated image quality of a tilted thick specimen by experiment and analysis. In a 3 MV ultra-HVEM, we obtained transmission electron images in amplitude contrast of 100 nm gold particles on the top surface of a tilted 5 μm thick amorphous epoxy-resin film. From line profiles of the images, we then measured and evaluated image blurring, contrast, and the signal-to-noise ratio (SNR) under different effective thicknesses of the tilted specimen and accelerating voltages of electrons. The variation of imaging blurring was consistent with the analysis based on multiple elastic scattering. When the effective thickness almost tripled, image blurring increased from ～3 to ～20 nm at the accelerating voltage of 3 MV. For the increase of accelerating voltage from 1 to 3 MV in the condition of the 14.6 μm effective thickness, due to the reduction of multiple scattering effects, image blurring decreased from ～54 to ～20 nm, and image contrast and SNR were both obviously enhanced by a factor of ～3 to preferable values. The specimen thickness was shown to influence image quality more than the accelerating voltage. Moreover, improvement on image quality of thick specimens due to increasing the accelerating voltage would become less when it was further increased from 2 to 3 MV in this work.     

Morphological characterisation of vesicular structures in the canine ejaculate

Membrane vesicles (MV) have been identified in seminal plasma from various species and they are thought to have a significant impact on semen quality and fertilisation. Although recently presence of MV has been also described in the canine ejaculate, detailed knowledge on their morphology is missing by now. This is, however, needed to provide a basis for detailed biochemical and functional studies as it is generally assumed that different MV populations are responsible for distinct tasks. MV were prepared for light (LM) and transmission electron microscopy (TEM) analysis using samples from normospermic dogs (n = 15), hypokinozoospermic dogs (n = 2, h) and one castrated azoospermic dog (a). For TEM, a new preparation protocol was used resulting in a higher MV retrieval rate. Using fractionated semen samples, most MV were identified in the second (sperm-rich) fraction in LM. Using pooled ejaculates, three different MV types could be identified in LM: (1) large MV with a marginal accumulation of opaque, granulated material, (2) medium- to small size MV with dense, opaque content and (3) small MV with no further defined contents. No direct contact between sperm and MV could be visualised. In TEM, 11 different MV types were identified based on diameter, structure, contents and electron density of contents as well as presence, number and size of smaller MV inside the MV itself. In normospermic males, secondary vesicles (type i, H, K1/2) included smaller vesicles and had a weighted mean diameter of 409.46 nm; hereof types i, H and K1 were smaller (mean: 287.55 nm, range: 51.25–994.86 nm) and type K2 was larger (mean: 1746.43 nm, range: 1003.66–3289.34 nm). Primary vesicles (mean diameter: 135.29 nm) – without vesicles inside – were differentiated into larger MV (A, B, C1/2) with a mean diameter of 219.63 nm (range: 39.08–1300.13 nm) and small primary MV (F, G) with a mean diameter of 66.12 nm (range: 24.62–99.84 nm). Whereas all mentioned MV were round to oval and mostly double-, rarely multiple-membrane surrounded, one longish primary MV type (L) was identified. In general, small primary vesicles were most common independent of semen quality, but distribution frequency of vesicle types differed between normospermic, pathospermic dogs and the castrated male. Mean weighted diameter of MV was 195.14 nm (range: 24.62–3289.34 nm) in normospermic males with the maximum diameter being smaller in the other dogs (h: 2096.78 nm; a: 1314.06 nm). Our results provide new information about ultrastructure and distribution frequency of canine MV in normospermic males and point to possible differences in MVs depending on semen quality. They provide the basis for further detailed functional analysis of MV subpopulations. Furthermore, the presence of MV in the castrated azoospermic male confirms an at least partly prostatic origin of canine MV.     

Soft X-ray imaging of thick carbon-based materials using the normal incidence multilayer optics

The high transparency of carbon-containing materials in the spectral region of “carbon window” (λ ～ 4.5–5 nm) introduces new opportunities for various soft X-ray microscopy applications. The development of efficient multilayer coated X-ray optics operating at the wavelengths of about 4.5 nm has stimulated a series of our imaging experiments to study thick biological and synthetic objects. Our experimental set-up consisted of a laser plasma X-ray source generated with the 2nd harmonics of Nd–glass laser, scandium-based thin-film filters, Co/C multilayer mirror and X-ray film UF-4. All soft X-ray images were produced with a single nanosecond exposure and demonstrated appropriate absorption contrast and detector-limited spatial resolution. A special attention was paid to the 3D imaging of thick low-density foam materials to be used in design of laser fusion targets.     

Automatic coarse-alignment for TEM tilt series of rod-shaped specimens collected with a full angular range

An automatic coarse-alignment method for a tilt series of rod-shaped specimen collected with a full angular range (from α = ?90° to +90°, α is the tilt angle of the specimen) is presented; this method is based on a cross-correlation method and uses the outline of the specimen shape. Both the rotational angle of the tilt axis and translational value of each image can be detected in the images without the use of markers. This method is performed on the basis of the assumption that the images taken at α = ?90° and α =  + 90° are symmetric about the tilt axis. In this study, a carbon rod on which gold particles have been deposited is used as a test specimen for the demonstration. This method can be used as an automatic coarse-alignment method prior to the application of a highly accurate alignment method because the alignment procedure can be performed automatically except for the initial setup of some parameters.     

Application of optical 3D measurement on thin film buckling to estimate interfacial toughness

The shape-from-focus (SFF) method has been widely studied as a passive depth recovery and 3D reconstruction method for digital images. An important step in SFF is the calculation of the focus level for different points in an image by using a focus measure. In this work, an image entropy-based focus measure is introduced into the SFF method to measure the 3D buckling morphology of an aluminum film on a polymethylmethacrylate (PMMA) substrate at a micro scale. Spontaneous film wrinkles and telephone-cord wrinkles are investigated after the deposition of a 300 nm thick aluminum film onto the PMMA substrate. Spontaneous buckling is driven by the highly compressive stress generated in the Al film during the deposition process. The interfacial toughness between metal films and substrates is an important parameter for the reliability of the film/substrate system. The height profiles of different sections across the telephone-cord wrinkle can be considered a straight-sided model with uniform width and height or a pinned circular model that has a delamination region characterized by a sequence of connected sectors. Furthermore, the telephone-cord geometry of the thin film can be used to calculate interfacial toughness. The instability of the finite element model is introduced to fit the buckling morphology obtained by SFF. The interfacial toughness is determined to be 0.203 J/m² at a 70.4° phase angle from the straight-sided model and 0.105 J/m² at 76.9° from the pinned circular model.     

FPGA implementation of real-time SENSE reconstruction using pre-scan and Emaps sensitivities

Sensitivity Encoding (SENSE) is a widely used technique in Parallel Magnetic Resonance Imaging (MRI) to reduce scan time. Reconfigurable hardware based architecture for SENSE can potentially provide image reconstruction with much less computation time. Application specific hardware platform for SENSE may dramatically increase the power efficiency of the system and can decrease the execution time to obtain MR images. A new implementation of SENSE on Field Programmable Gate Array (FPGA) is presented in this study, which provides real-time SENSE reconstruction right on the receiver coil data acquisition system with no need to transfer the raw data to the MRI server, thereby minimizing the transmission noise and memory usage. The proposed SENSE architecture can reconstruct MR images using receiver coil sensitivity maps obtained using pre-scan and eigenvector (E-maps) methods. The results show that the proposed system consumes remarkably less computation time for SENSE reconstruction, i.e., 0.164 ms @ 200 MHz, while maintaining the quality of the reconstructed images with good mean SNR (29 + dB), less RMSE (< 5 × 10^− 2) and comparable artefact power (< 9 × 10^− 4) to conventional SENSE reconstruction. A comparison of the center line profiles of the reconstructed and reference images also indicates a good quality of the reconstructed images. Furthermore, the results indicate that the proposed architectural design can prove to be a significant tool for SENSE reconstruction in modern MRI scanners and its low power consumption feature can be remarkable for portable MRI scanners.     

Discrimination methods for biological contaminants in fresh-cut lettuce based on VNIR and NIR hyperspectral imaging

The rapid detection of biological contaminants such as worms in fresh-cut vegetables is necessary to improve the efficiency of visual inspections carried out by workers. Multispectral imaging algorithms were developed using visible-near-infrared (VNIR) and near-infrared (NIR) hyperspectral imaging (HSI) techniques to detect worms in fresh-cut lettuce. The optimal wavebands that can detect worms in fresh-cut lettuce were investigated for each type of HSI using one-way ANOVA. Worm-detection imaging algorithms for VNIR and NIR imaging exhibited prediction accuracies of 97.00% (RI_547/945) and 100.0% (RI_1064/1176, SI_1064-1176, RSI-I_{(1064-1173)/1064}, and RSI-II_{(1064-1176)/(1064+1176)})_, respectively. The two HSI techniques revealed that spectral images with a pixel size of 1 × 1 mm or 2 × 2 mm had the best classification accuracy for worms. The results demonstrate that hyperspectral reflectance imaging techniques have the potential to detect worms in fresh-cut lettuce. Future research relating to this work will focus on a real-time sorting system for lettuce that can simultaneously detect various defects such as browning, worms, and slugs.     

Electron penetration depth in amorphous AlN exploiting the luminescence of AlN:Tm/AlN:Ho bilayers

The penetration depth of electron in amorphous aluminum nitride (AlN) is determined in terms of energy loss per unit length using electron beam in a cathodoluminescence (CL) apparatus. Thin films bilayers of holmium doped aluminum nitride (AlN:Ho) and thulium doped aluminum nitride (AlN:Tm) are deposited on silicon substrates by rf magnetron sputtering method at liquid nitrogen temperatures. The bilayers structure consisted of a 37.8 nm thick AlN:Tm film on the top of a 15.3 nm thick AlN:Ho film. Electron beam of different energies are allowed to penetrate the AlN:Tm/AlN:Ho bilayers film. The spectroscopic properties of AlN:Ho and AlN:Tm, the thickness of the film and the energies of electron beam are used to calculate the penetration depth of electron in amorphous AlN. Electron beam of 2.5 keV energy was able to pass through the 37.8 nm thick AlN:Tm film. The electron penetration depth for AlN is found to be 661.4 MeV/cm.     

Preparation of cadmium stannate films by spray pyrolysis technique

Cadmium stannate thin films were prepared by spray pyrolysis technique using cadmium acetate and tin(II) chloride precursors at substrate temperatures 450 °C and 500 °C. XRD pattern confirms the formation of orthorhombic (1 1 1) cadmium stannate phase for the film prepared at substrate temperature of 500 °C, whereas, films prepared at 450 °C are amorphous. Film formation does not occur at substrate temperature from 300 to 375 °C. SEM images reveal that the surface of the prepared Cd₂SnO₄ film is smooth. The average optical transmittance of ∼86% is obtained for the film prepared at substrate temperature of 500 °C with the film thickness of 400 nm. The optical band gap value of the films varies from 2.7 to 2.94 eV. The film prepared at 500 °C shows a minimum resistivity of 35.6 × 10⁻⁴ Ω cm.     

Classifying of Nellore cattle beef on Normal and DFD applying a non conventional technique

The aim of this study was to evaluate the accuracy in the Normal and DFD classification in Nellore beef using a bench-top hyperspectral imaging system. A hyperspectral imaging system (λ = 928–2524 nm) was used to collect hyperspectral images of the Longissimus thoracis et lumborum (n = 78) of Nellore cattle. The images were processed, being selected region of interest and extracted spectra image and were selected the wavelengths considered most important for the treats evaluated. Six linear discriminant models were developed to classify beef samples on Normal and DFD. The model using all wavelengths associated with the reflectance and absorbance spectrum transformed with the pretreatment 2nd derivative resulted in an overall accuracy of 93.6% for both pretreatments. In this configuration, the model was able to classify correctly 73 samples from a total of 78 samples. The results demonstrate that the hyperspectral imaging system may be considered a viable technology for beef classification on Normal and DFD.     

Compressed sensing based simultaneous black- and gray-blood carotid vessel wall MR imaging

ObjectiveIn this study, we sought to demonstrate the blood suppression performance, image quality and morphological measurements for compressed sensing (CS) based simultaneous 3D black- and gray-blood imaging sequence (CS-siBLAG) in carotid vessel wall MR imaging.Materials and methodsSeven healthy volunteers and five patients were recruited. Healthy subjects underwent five CS-siBLAG scans with 1, 2, 3, 4 and 5-fold accelerations. Signal-to-tissue ratio (STR) and contrast-to-tissue ratio (CTR) were computed as the measures of flowing signal suppression performance and the image quality for black-blood imaging of the technique. Vessel lumen area (LA) and wall area (WA) were compared between fully sampled acquisition and each accelerated acquisition. Patients underwent three CS-siBLAG scans with 1, 3 and 5-fold accelerations as well as a 3D time of flight (3D TOF) scan. Two radiologists reviewed the under-sampled black- and gray-blood image quality.ResultsSTR and CTR values obtained with 2 to 5-fold accelerations were not significantly different from those with full acquisition. LA and WA measured at 2 ×, 3 ×, 4 × and 5 × were all highly correlated to the corresponding values at 1 ×. For patients imaging, two radiologists both found that the dual-contrast images at 3 × acceleration exhibited comparable image quality to that of the fully sampled acquisition, and that the images at 5 × exhibited slightly blurred vessel wall and outer vessel wall boundaries.ConclusionBy combining the CS under-sampling pattern and reconstruction, pseudo-centric phase encoding order and dual blood contrast sequences, this technique provides spatially registered black- and gray-blood images and excellent visualization for vessel wall imaging and gray-blood imaging in a short scan time.     

LEED and STM studies of the stability of the MoO2(100) surface

Atomic scale images and low energy electron diffraction pattern of a MoO₂(100) single crystal surface are presented, which show different structural modifications depending on surface preparation. A short in-situ heat treatment of the as-grown single crystal results in an atomically ordered surface whose diffraction pattern and STM images are consistent with those expected from the bulk structure. The symmetry of the STM images suggests an oxygen termination of the surface. A significantly longer heat treatment causes a thermodynamically stable (4 × 1) reconstruction which is interpreted to be due to a loss of oxygen chains. The (4 × 1) reconstruction vanishes after Ar-ion-sputtering and subsequent annealing. Additional long sputtering cycles result in a (2 × 1) reconstruction. The observed surface reconstructions can be transformed into each other by heating or sputtering cycles.     

Simulation and measurement of AES depth profiles; a case study of the C/Ta/C/Si system

A multilayer sample (C (23.3 nm)/Ta (26.5 nm)/C (22.7 nm)/Si substrate) was submitted to AES depth profiling by Ar⁺ ions of energy 1 keV and angles of incidence of 72°, 78°, and 82°. The shapes of the as-measured depth profiles were strongly different emphasizing that the ion-bombardment conditions strongly affects the shapes of measured depth profiles. We simulated the depth profile measured at an angle of incidence of 72° by calculating the backscattering factor, applying attenuation lengths available in the literature, and simulating the ion-bombardment-induced specimen alteration with a TRIDYN simulation and a trial and error method. The good agreement between the calculated and measured depth profiles justified the method applied.     

Maintaining the genuine structure of 2D materials and catalytic nanoparticles at atomic resolution

The recent development of atomic resolution, low dose-rate electron microscopy allows investigating 2D materials as well as catalytic nano particles without compromising their structural integrity. For graphene and a variety of nanoparticle compositions, it is shown that a critical dose rate exists of <100 e⁻/Ų s at 80 keV of electron acceleration that allows maintaining the genuine object structures including their surfaces and edges even if particles are only 3 nm large or smaller. Moreover, it is demonstrated that electron beam-induced phonon excitation from outside the field of view contributes to a contrast degradation in recorded images. These degradation effects can be eliminated by delivering electrons onto the imaged area, only, by using a Nilsonian illumination scheme in combination with a suitable aperture at the electron gun/monochromator assembly.     

Simultaneous two color image capture for sub-diffraction localization fluorescence microscopy

A sub-diffraction limit fluorescence localization microscope was constructed using a standard cooled 1.4 mega-pixel fluorescence charge-coupled device (CCD) camera to simultaneously resolve closely adjacent paired quantum dots on a flat surface with emissions of 540 and 630 nm. The images of the overlapping Airy discs were analyzed to determine the center of the point spread function after noise reduction using Fourier transformation analysis. The Cartesian coordinates of the centers of the point spread functions were compared in serial images. Histograms constructed from serial images fit well to Gaussian functions for resolving two quantum dots separated by as little as 10 nm in the x–y coordinates. Statistical analysis of multiple pairs validated discrimination of inter-fluorophore distances that vary by 10 nm. The method is simple and developed for x–y resolution of dilute fluorophores on a flat surface, not serial z sectioning.     

Interface strain profiling in ultrathin SiO2 gate oxides with medium energy ion scattering spectroscopy

Medium energy ion scattering spectroscopy (MEIS) could identify ∼1 nm interface layer with compressive strain, which depends sensitively on the interface treatment conditions such as oxynitridation, ozone oxidation, tilt of Si(0 0 1) substrates. The interface strain relaxation always shows improvements in gate oxide reliability. Atomic scale investigations of strain profiles with MEIS are reviewed for SiO₂/Si(0 0 1) interfaces.     

Hardness and microstructural variation of Al–Mg–Mn–Sc–Zr alloy

Variations of Vickers hardness were observed in Al–Mg–Mn alloy and Al–Mg–Mn–Sc–Zr alloy at different ageing times, ranging from a peak value of 81.2 HV at 54 ks down to 67.4 HV at 360 ks, below the initial hardness value, 71.8 HV at 0 ks for the case of Al–Mg–Mn–Sc–Zr alloy. Microstructures of samples at each ageing stage were examined carefully by transmission electron microscopes (TEMs) both in two-dimensions and three-dimensions. The presence of different types, densities, and sizes of particles were observed dispersed spherical Al₃Sc_1−xZr_x and also block-shaped Al₃Sc precipitates growing along <1 0 0>_Al with facets {1 0 0} and {1 1 0} of the precipitates. TEM analysis both in two-dimensions and three-dimensions, performed on various samples, confirmed the direct correlation between the hardness and the density of Al₃Sc.