Dual-color 4Pi-confocal microscopy with 3D-resolution in the 100 nm range

We report the development of simultaneous two-color channel recording in 4Pi-confocal microscopy. A marked increase of spatial resolution over confocal microscopy becomes manifested in 4Pi-confocal three-dimensional (3D) data stacks of dual-labeled objects. The fundamentally improved resolution is verified both with densely labeled fluorescence beads as well as with membrane labeled fixed Escherichia coli. The synergistic combination of dual-color 4Pi-confocal recording with image restoration results in dual-color imaging with a 3D resolution in the 100 nm range.     

Evaluation of Nanoscale Friction Depth Distribution in ZDDP and MoDTC Tribochemical Reacted Films Using a Nanoscratch Method

The distributions of local friction coefficients relative to the depth and near the surface of MoDTC/ZDDP and ZDDP tribofilms were successfully evaluated by using a nanoscratch method combined with in situ AFM observation. It was found that both tribofilms were friction-functionally graded materials. The friction coefficients decreased from 0.35 to 0.16 with a decrease in the scratch depth from 60 to 10 nm. It was observed that the MoDTC/ZDDP and ZDDP tribofilms possessed different shear strength levels near the surface as evidenced by the different valley-shaped friction coefficient distributions they exhibited for scratch depths ranging from 2 to 10 nm. Based on our recent nanomechanical measurements, this observation indicated that both tribofilms possessed an ultra-low friction inner skin layer at a depth of about 10 nm below the surface. Most importantly, the inner skin layer of the MoDTC/ZDDP tribofilm possessed a lower friction coefficient than that of the ZDDP tribofilm (0.084 versus 0.104) and was thinner (about 3.2 nm versus 6.4 nm). These results thus revealed that the reduction in friction attributed to the MoDTC additive originates from the different friction behavior of the inner skin layers of the MoDTC/ZDDP and ZDDP tribofilms. These nanoscratch results agree with the findings of our recent work on detecting differences in mechanical properties between these tribofilms by nanoindentation measurements.     

Evaluation of Local Mechanical Properties in Depth in MoDTC/ZDDP and ZDDP Tribochemical Reacted Films Using Nanoindentation

Local mechanical properties in depth and near the surface of MoDTC/ZDDP and ZDDP tribofilms, which exhibited obviously different friction coefficients in a pin-on-disc test, were determined by using a nanoindentation technique combined with in-situ atomic force microscopy (AFM) observation. Tapping-mode AFM observation revealed that the MoDTC/ZDDP film was much rougher than the ZDDP film. Nanoindentation measurement revealed that the MoDTC/ZDDP and ZDDP tribofilms possessed different elasto-plasticities around a depth of several nanometers from the surface, although both films showed the same hardness and modulus depth distributions except in the surface area. The same mechanical depth distributions indicated that both kinds of tribofilm were functionally graded materials; that is, they consisted of a layer near the surface with lower hardness and modulus and providing lubrication and a base layer with higher hardness and modulus and serving to modify property differences at the interface. Most importantly, the different elasto-plasticities near the tribofilm surfaces revealed that the MoDTC/ZDDP tribofilm possessed lower shearing yield stress than the ZDDP tribofilm. The results of this study suggest that the presence of some solid lubricants such as MoS₂ just below the MoDTC/ZDDP film surface reduced the boundary friction coefficient.     

Development of a new model for representing elastoplastic deformation and fracture behaviors of solid fats

A new simulation model, namely, ADEM Ductile Fracture model (ADF model) is developed to represent elastoplastic deformation and fracture behaviors of solid fats. Experimental measurement of the coefficient of plastic starting distance and the yield point affecting the deformation behavior has been obtained by compression test. The fracture distance affecting the fracture behavior has been measured by cutting test. The ADF model shows a good match with the experimentally determined compression and cutting behaviors of the solid fats, and load–strain curves as a function of plate thickness. This confirms that the ADF model could represent the elastoplastic deformation and fracture behaviors of the solid fats.     

Supercontinuum dynamically visualizes a dividing single cell

During cell division, various organelles behave dynamically. Visualization of these dynamic behaviors of organelles is a promising one step forward for understanding life at the molecular level. One- or two-photon excited fluorescence microscopy has so far been used for visualizing these cell dynamics. The fluorescent probe introduced into a living cell can visualize the spatial distribution of a target molecule in real time, enabling the tracing of cell dynamics at the molecular level. Introducing a fluorescent probe into a cell, however, may alter the physical and chemical conditions of the cell. Here we show a new method for direct (no need for staining cells) visualization of living cell processes with coherent anti-Stokes Raman scattering (CARS) spectroscopy. A new light source, supercontinuum generated from a photonic crystal fiber, has facilitated ultrabroadband (>3500 cm(-1)) multiplex CARS spectroscopy and imaging with high molecular specificity. Using this multiplex CARS technique, we have been successful in tracing the whole cell division process, the splitting of a mother cell into two daughter cells, appearance and disappearance of septum, and dynamic distribution changes of organelles consisting of lipid membrane. The supercontinuum has also facilitated simultaneous measurement of the CARS and two-photon excited fluorescence (TPEF) spectra, enabling what we call multiple nonlinear spectral imaging. Three-dimensional image reconstruction of a living cell with high speed is now possible to elucidate more detailed molecular-level dynamics inside a dividing living cell.     

Degradative capacities and bioaugmentation potential of an anaerobic benzene-degrading bacterium strain DN11

Azoarcus sp. strain DN11 is a denitrifying bacterium capable of benzene degradation under anaerobic conditions. The present study evaluated strain DN11 for its application to bioaugmentation of benzene-contaminated underground aquifers. Strain DN11 could grow on benzene, toluene, m-xylene, and benzoate as the sole carbon and energy sources under nitrate-reducing conditions, although o- and p-xylenes were transformed in the presence of toluene. Phenol was not utilized under anaerobic conditions. Kinetic analysis of anaerobic benzene degradation estimated its apparent affinity and inhibition constants to be 0.82 and 11 microM, respectively. Benzene-contaminated groundwater taken from a former coal-distillation plant site was anaerobically incubated in laboratory bottles and supplemented with either inorganic nutrients (nitrogen, phosphorus, and nitrate) alone, or the nutrients plus strain DN11, showing that benzene was significantly degraded only when DN11 was introduced. Denaturing gradient gel electrophoresis of PCR-amplified 16S rRNA gene fragments, and quantitative PCR revealed that DN11 decreased after benzene was degraded. Following the decrease in DN11 16S rRNA gene fragments corresponding to bacteria related to Owenweeksia hongkongensis and Pelotomaculum isophthalicum, appeared as strong bands, suggesting possible metabolic interactions in anaerobic benzene degradation. Results suggest that DN11 is potentially useful for degrading benzene that contaminates underground aquifers at relatively low concentrations.     

Saccharomyces cerevisiae mutant displaying beta-glucans on cell surface

The deletion of MCD4 leads to an increase in beta-1,6-glucan level and a decrease in glycosylphosphatidylinositol-anchored protein and mannan levels in the cell wall of Saccharomyces cerevisiae, suggesting that mcd4 deletion mutant (mcd4Delta) displays beta-glucans on the cell surface without a mannan cover. An observation of the cell surface of mcd4Delta cells and an examination of the effect of contact between mcd4Delta cells and mouse macrophages indicated that macrophages were activated by contact with mcd4Delta cells displaying beta-glucans on the cell surface. We further examined the effect of intraperitoneal ethanol-fixed mcd4Delta cells on the survival period of mice infected with Candida albicans. mcd4Delta cells prolonged the survival period, implying that mcd4Delta cells may enhance the immune function of mice via macrophage activation. Moreover, we examined the structures of beta-glucans (i.e., alkali- and acetic acid-insoluble beta-glucans) extracted from mcd4Delta with (13)C-NMR and the effect of extracted beta-glucans on TNF-alpha secretion from macrophages. The structures of the beta-glucans from mcd4Delta differed from those of wild type (WT); however, there was no difference in tumor necrosis factor-alpha (TNF-alpha) secretion level between beta-glucans from mcd4Delta and those from WT. The yield of purified beta-glucans obtained from dry cells of mcd4Delta was higher than that obtained from dry cells of WT. mcd4Delta may be a superior strain for the preparation of beta-glucans.     

Design investigation of a linear motor for artificial heart driving

The authors have been considering the use of a linear pulse motor (LPM) as a driving source of a totally implantable artificial heart (TAH), and have developed a series of artificial heart models that incorporate such a motor. The newly developed linear-type TAH (linear TAH) has two blood-pumps, which are inflated and deflated alternately by the LPM, causing a pusher plate to pulsate the blood. This paper deals with the performance calculation of the LPM using the loading ratio. Some motors manufactured for driving TAHs were then selected to compare their machine parameters. The selected motors were LPM, linear oscillatory actuator, brushless dc motors, and ultrasonic motor. Two parameters were selected for the comparison: the mechanical output/volume ratio and the motor constant. The results may be summarized as follows: 1. The LPM was designed based on the loading ratio, and the kinetic thrust and velocity were obtained at the range of 77 newtons and 80 mm/s, respectively. 2. The loading ratio of the LPMs were more than eight times larger in comparison with the rotary motors that were used in industrial machines. 3. The motor constants of the LPMs were recognized to be one figure larger than that of the brushless dc motors. In order to reduce the volume of the linear TAH, it is necessary to increase the drive velocity of the LPMs. Application of the linear motor to a nonpulsatile artificial heart will be one way to achieve this. © 1998 Scripta Technica, Electr Eng Jpn, 123(1): 43–50, 1998     

The effect of ZDDP in CVT fluid on increasing the traction capacity of belt-drive continuously variable transmissions

Belt-drive continuously variable transmissions are already being increasingly used in Europe and Japan for small vehicles, but midsize vehicles will require greater torque capacity. This paper discusses the effect of ZDDP as an additive on increasing the traction obtained at the sliding interfaces between the elements and pulleys of a belt-drive continuously variable transmission (B-CVT). This effect stems from the higher friction coefficient at the sliding interfaces. Friction coefficients were measured in rig tests in which test pieces cut from actual CVT elements were slid against a disc in different types of CVT fluid (CVTF). After the tests, the adsorbed films on the specimen surfaces were investigated with electron probe microanalysis (EPMA), and compared with pieces cut from elements subjected to a high torque test conducted with the B-CVT. The experimental results indicated that the higher friction coefficient at the sliding interfaces was due to the shearing of the tough adsorbed film. The film was blue in colour, and consisted mainly of zinc precipitated from the ZDDP additive.     

Stacking faults in an epitaxially grown PbTiO3 thick film and their size distribution

The microstructure of an epitaxial PbTiO₃ thick film, grown on a SrRuO₃/SrTiO₃ substrate at 600 °C by pulsed-MOCVD method, was investigated by using transmission electron microscopy. A number of extrinsic or intrinsic stacking faults were observed in the epitaxial PbTiO₃ thick film and they were parallel to the (0 0 1) plane of the PbTiO₃. We also investigated the size distribution of these stacking faults. The width of these stacking faults along the [1 0 0] axis of the PbTiO₃ was very small, ranging from 2 to 13 nm. It was also revealed that the size distribution of stacking faults depends on the position in the film: near the surface, near the substrate, near threading dislocations, and near 90° domain boundaries.