An integrated approach to the study of living cells by atomic force microscopy

We describe a technique for studying living cells with the atomic force microscope (AFM) in tapping mode using a thermostated, controlled-environment culture system. We also describe the integration of the AFM with bright field, epifluorescence and surface interference microscopy, achieving the highest level of integration for the AFM thus far described. We succeeded in the continuous, long-term imaging of relatively flat but very fragile cytoplasmic regions of COS cells at a lateral resolution of about 70 nm and a vertical resolution of about 3 nm. In addition, we demonstrate the applicability of our technology for continuous force volume imaging of cultured vertebrate cells.
The hybrid instrument we describe can be used to collect simultaneously a diverse variety of physical, chemical and morphological data on living vertebrate cells. The integration of light microscopy with AFM and steady-state culture methods for vertebrate cells represents a new approach for studies in cell biology and physiology.     

Quantitative structural analysis of twin boundary in alpha-Zn7Sb2O12 using HAADF STEM method

The structure and composition of the 1/4{110} twin boundary in alpha-Zn7Sb2O12 have been determined by using quantitative high-angle annular dark field scanning transmission electron microscopy (HAADF STEM) analysis. The noise in the experimental HAADF STEM images is reduced by using the maximum entropy method and average processing, and the parameters used in dynamical simulations are experimentally determined. From the analysis, it has been found that octahedral sites in the twin boundary slightly shift parallel to the [110] direction, and a reduction of the Sb concentration at the octahedral sites on the plane adjacent to the twin boundary was detected. The reduction was measured from three regions in the same twin boundary, and the Sb concentrations were 4 +/- 3, 8 +/- 3 and 19 +/-2 at% from 33 at%.     

Measurement of axial neutral density profiles in a microwave discharge ion thruster by laser absorption spectroscopy with optical fiber probes

In order to reveal the physical processes taking place within the "μ10" microwave discharge ion thruster, internal plasma diagnosis is indispensable. However, the ability of metallic probes to access microwave plasmas biased at a high voltage is limited from the standpoints of the disturbance created in the electric field and electrical isolation. In this study, the axial density profiles of excited neutral xenon were successfully measured under ion beam acceleration by using a novel laser absorption spectroscopy system. The target of the measurement was metastable Xe I 5p(5)((2)P(0) (3/2))6s[3/2](0) (2) which absorbed a wavelength of 823.16 nm. Signals from laser absorption spectroscopy that swept a single-mode optical fiber probe along the line of sight were differentiated and converted into axial number densities of the metastable neutral particles in the plasma source. These measurements revealed a 10(18) m(-3) order of metastable neutral particles situated in the waveguide, which caused two different modes during the operation of the μ10 thruster. This paper reports a novel spectroscopic measurement system with axial resolution for microwave plasma sources utilizing optical fiber probes.     

Bright-field scanning confocal electron microscopy using a double aberration-corrected transmission electron microscope

Scanning confocal electron microscopy (SCEM) offers a mechanism for three-dimensional imaging of materials, which makes use of the reduced depth of field in an aberration-corrected transmission electron microscope. The simplest configuration of SCEM is the bright-field mode. In this paper we present experimental data and simulations showing the form of bright-field SCEM images. We show that the depth dependence of the three-dimensional image can be explained in terms of two-dimensional images formed in the detector plane. For a crystalline sample, this so-called probe image is shown to be similar to a conventional diffraction pattern. Experimental results and simulations show how the diffracted probes in this image are elongated in thicker crystals and the use of this elongation to estimate sample thickness is explored.     

Experimental study on hysteresis phenomena of shock wave structure in an over-expanded axisymmetric jet

In recent studies on two-dimensional supersonic jets, it is reported that the hysteresis phenomenon for the reflection type of shock waves in the jet flow field is occurred under the quasi-steady flow condition and this phenomenon is affected by the transitional pressure ratio between the regular reflection and Mach reflection. However, so far, there are few researches on the hysteresis phenomenon for the transition of shock waves between regular and Mach reflection in over-expanded supersonic jets and the phenomenon has not been investigated satisfactorily. Therefore, the purpose of this study is to clarify the hysteresis phenomena for the reflection type of shock wave in the over-expanded axi-symmetric supersonic jet experimentally, and to discuss the relationship between hysteresis phenomenon and rate of the change of pressure ratio with time. Furthermore, the effect of Mach number at the nozzle exit on hysteresis loop was investigated for two kinds of nozzle.     

Real‐time histological imaging of kidneys stained with food dyes using multiphoton microscopy

We have developed a real‐time imaging technique for diagnosis of kidney diseases which is composed of two steps, staining renal cells safely with food dyes and optical sectioning of living renal tissue to obtain histological images by multiphoton microscopy (MPM). Here, we demonstrated that the MPM imaging with food dyes, including erythrosine and indigo carmine, could be used as fluorescent agents to visualize renal functions and structures such as glomerular bloodstreams, glomerular filtration, and morphology of glomeruli and renal tubules. We also showed that the kidneys of IgA nephropathy model‐mice stained with the food dyes presented histopathological characteristics different from those observed in normal kidneys. The use of the food dyes enhances the quality of tissue images obtained by MPM and offers the potential to contribute to a clinical real‐time diagnosis of kidney diseases. Microsc. Res. Tech. 78:847–858, 2015. © 2015 Wiley Periodicals, Inc.     

Sol–Gel‐Derived High‐Performance Stacked Transparent Conductive Oxide Thin Films

Single‐ and multi‐layer transparent conductive oxide (TCO) thin films exhibiting high performance, good packing density and low surface/interface roughness are deposited on silica glass substrates by the sol–gel method. The crystal and microstructural properties of the TCO thin films are evaluated as an alternate to films prepared by ultra‐high vacuum deposition. Tin‐doped indium oxide (ITO) thin films produced using a two‐step drying process showed low surface roughness because of dense packing structure not only horizontal but also vertical directions. As a result, electrical conductivity, carrier concentration, carrier mobility, and optical transmittance of 2.3 × 10³ S/cm, 8 × 10²⁰ cm^?3, 18 cm²/Vs, and over 98% at 500 nm, respectively, were achieved. A multilayer ZnO/ITO stacked structure was also fabricated using the sol–gel process. Our findings suggest that solution‐based methods show promise as an alternative to existing ultra‐high vacuum methods to fabricate TCO thin films.     

Enrichment of CLA isomers by selective esterification with l-menthol using Candida rugosa lipase

Commercially available preparations of CLA are composed of almost equal amounts of 9-cis,11-trans (9c,11t)-CLA and 10-trans,12-cis (10t,12c)-CLA. Each isomer was fractionated and enriched, for availability as a food supplement, by a process comprising selective esterification with l-menthol by Candida rugosa lipase, distillation, and n-hexane extraction. The first selective esterification of CLA isomers was conducted with an equimolar amount of l-menthol of 30°C. The oil phase of the reaction mixture was fractionated into an l-menthyl ester fraction (9c,11t-CLA rich) and an FFA fraction (10t,12c-CLA rich) by distillation. The FFA fraction was esterified again with an equimolar amount of l-menthol to enrich 10t,12c-CLA. The 10t,12c-CLA preparation was obtained as the resulting FFA fraction by distillation. 10t,12c-CLA was enriched to 91% with 40% recovery. To enrich 9c,11t-CLA, the l-menthyl ester fraction in the first esterification was chemically hydrolyzed, and the resulting FFA were esterified again with an equimolar amount of l-menthol. The 9c, 11t-CLA preparation was obtained by chemical hydrolysis of the resulting l-methyl ester fraction, followed by n-hexane extraction. 9c,11t-CLA was enriched to 94% with 42% recovery. This effective process for purification of CLA isomers using l-methol is applicable to the production of food supplements.     

Development of a two-dimensional dual pendulum thrust stand for Hall thrusters

A two-dimensional dual pendulum thrust stand was developed to measure thrust vectors [axial and horizontal (transverse) direction thrusts] of a Hall thruster. A thruster with a steering mechanism is mounted on the inner pendulum, and thrust is measured from the displacement between inner and outer pendulums, by which a thermal drift effect is canceled out. Two crossover knife-edges support each pendulum arm: one is set on the other at a right angle. They enable the pendulums to swing in two directions. Thrust calibration using a pulley and weight system showed that the measurement errors were less than 0.25 mN (1.4%) in the main thrust direction and 0.09 mN (1.4%) in its transverse direction. The thrust angle of the thrust vector was measured with the stand using the thruster. Consequently, a vector deviation from the main thrust direction of +/-2.3 degrees was measured with the error of +/-0.2 degrees under the typical operating conditions for the thruster.     

Thermal stress measurement in steel plate strengthened by CFRP and aluminum alloy plates

In steel members strengthened by carbon-fiber reinforced polymer (CFRP) plates, the thermal stresses are introduced in the steel members, the CFRP plates and the adhesive layers when temperature changes because the linear thermal expansion coefficients of steel and CFRP are mismatched. As so far, the authors proposed a technique to reduce the thermal stress in steel members strengthened by CFRP plates, which involves bonding aluminum alloy plates with CFRP plates. In the proposed method, the thermal stress in steel member can be reduced so that there are negligible levels of stress in steel member when the cross sectional areas of CFRP and aluminum plates are designed to correspond to the coefficient of thermal expansion of steel, even though the thermal stresses are introduced in the CFRP and aluminum plates. In this study, to confirm the maintaining the thermal stress reduction in steel member by proposed method, thermal stress measurement in steel plate strengthened by CFRP and aluminum plates was carried out about 21 months. In this research, the thermal stress introduced in the steel plate strengthened by CFRP plates was also measured. Furthermore, to assume the thermal shear and normal (peel) stresses in adhesive layers, FE analysis with plane stress element was employed. As the result, it was shown the thermal stresses in steel plate with CFRP plate were able to calculate by using composite theory and measured temperature. Furthermore, in steel plate strengthened by CFRP and aluminum plates, the thermal stress introduced in steel plate was negligible-small through the all-season. It was found the thermal stresses in steel plate with CFRP plates as well as CFRP and aluminum plates were also estimated by using composite theory and measured temperature. In the steel plate strengthened by CFRP and aluminum plates, the thermal shear and normal stresses in adhesive layer glued to steel plate become smaller than that in the conventional CFRP bonded specimen. However, the shear stress in adhesive layers between CFRP and aluminum plates in proposed method was higher than the thermal stress in adhesive layers between CFRP plates in conventional method.