Distribution of nitric oxide synthase immunoreactivity in the mouse brain

Nitric oxide (NO) is a gaseous intercellular messenger with a wide range of neural functions. NO is synthesized by activation of different isoforms of nitric oxide synthases (NOS). At present NOS immunoreactivity has been described in mouse brain in restricted and definite areas and no detailed mapping studies have yet been reported for NOS immunoreactivity. We have studied the distribution of neuronal NOS-containing neurons in the brain of three months male mice, using a specific commercial polyclonal antibody against the neuronal isoform of nitric oxide synthase (nNOS). Neuronal cell bodies exhibiting nNOS immunoreactivity were found in several distinct nuclei throughout the brain. The neurons that were positively stained exhibited different intensities of reaction. In some brain areas (i.e., cortex, striatum, tegmental nuclei) neurons were intensely stained in a Golgi-like fashion. In other regions, immunoreactive cells are moderately stained (i.e., magnocellular nucleus of the posterior commissure, amygdaloid nucleus, interpeduncular nucleus, lateral periaqueductal gray) or weakly stained (i.e., vascular organ of the lamina terminalis, hippocampus, inferior colliculus, reticular nucleus). In the mouse, the NO-producing system appears well developed and widely diffused. In particular, nNOS immunoreactive neurons seem chiefly present in several sensory pathways like all the nuclei of the olfactory system, as well as in many regions of the lymbic system. These data suggest a widespread role for the NO system in the mouse nervous system.     

Plasmon energy mapping in energy-filtering transmission electron microscopy

In this paper, we demonstrate the two-dimensional mapping of plasmon energies by energy-filtering transmission electron microscopy. The maps are obtained from a series of energy-filtered images in the plasmon energy region. Examples are shown for a nano-crystalline Si-B-C-N ceramic. This material contains SiC and Si(3)N(4) grains as well as intergranular regions composed of hexagonal BN (h-BN) and turbostratic carbon (t-C). The different phases can be clearly identified by their specific plasmon energies. An energy resolution of < or =0.1eV is achieved. In addition, the plasmon map of an amorphous carbon film is used to visualize the non-isochromaticity of the Corrected Omega filter (90 degrees filter) of the SESAM2. A procedure is proposed for the correction of the non-isochromaticity.     

Theory experiment comparison of the electron backscattering factor from solids at low electron energy (250-5,000 eV)

The electron backscattering factor was measured from 24 different elements at low primary beam energy (250-5,000 eV). The results were compared with Monte Carlo simulations from a variety of freely available programs and an in-house developed program. The results suggest that a thin film of oxide can modify the backscattering factor at low primary energy. In addition, a number of problems have been identified with the freely available programs.     

The Thomson deflectometer: a novel use of the Thomson spectrometer as a transient field and plasma diagnostic

Laser accelerated proton beams have been used for field characterization in expanding plasmas. The Thomson parabola spectrometer, as a charged particles analyzer, also allows precise measurement of the charged particles' trajectories. The proton's deflections by fast changing plasma fields can be measured with the new design of the Thomson parabola spectrometer and, therefore, it can be applied for proton deflectometry. It is shown that from resulting spectrograms the plasma field dynamics can be reconstructed with high temporal resolution. In a proof-of-principle experiment, a weakly relativistic plasma expansion is studied as an example.     

Precession technique and electron diffractometry as new tools for crystal structure analysis and chemical bonding determination

We have developed a new fast electron diffractometer working with high dynamic range and linearity for crystal structure determinations. Electron diffraction (ED) patterns can be scanned serially in front of a Faraday cage detector; the total measurement time for several hundred ED reflections can be tens of seconds having high statistical accuracy for all measured intensities (1-2%). This new tool can be installed to any type of TEM without any column modification and is linked to a specially developed electron beam precession "Spinning Star" system. Precession of the electron beam (Vincent-Midgley technique) reduces dynamical effects allowing also use of accurate intensities for crystal structure analysis. We describe the technical characteristics of this new tool together with the first experimental results. Accurate measurement of electron diffraction intensities by electron diffractometer opens new possibilities not only for revealing unknown structures, but also for electrostatic potential determination and chemical bonding investigation. As an example, we present detailed atomic bonding information of CaF(2) as revealed for the first time by precise electron diffractometry.     

Surface damage induced by FIB milling and imaging of biological samples is controllable

Focused ion beam (FIB) techniques are among the most important tools for the nanostructuring of surfaces. We used the FIB/SEM (scanning electron microscope) for milling and imaging of digestive gland cells. The aim of our study was to document the interactions of FIB with the surface of the biological sample during FIB investigation, to identify the classes of artifacts, and to test procedures that could induce the quality of FIB milled sections by reducing the artifacts. The digestive gland cells were prepared for conventional SEM. During FIB/SEM operation we induced and enhanced artifacts. The results show that FIB operation on biological tissue affected the area of the sample where ion beam was rastering. We describe the FIB-induced surface major artifacts as a melting-like effect, sweating-like effect, morphological deformations, and gallium (Ga(+)) implantation. The FIB induced surface artifacts caused by incident Ga(+) ions were reduced by the application of a protective platinum strip on the surface exposed to the beam and by a suitable selection of operation protocol. We recommend the same sample preparation methods, FIB protocol for milling and imaging to be used also for other biological samples.     

A two-degree-of-freedom motor-powered gait orthosis for spinal cord injury patients

A number of orthoses have been developed to restore stance and walking in paraplegic subjects. Compliance, however, has been limited, mainly owing to walking effort. Use of the forces produced by actuators is an effective way to solve the problem of the considerable effort required for orthotic gait, namely high muscular effort and high energy expenditure. The purpose of the present study was to investigate the effects of assistance by external actuators on the orthotic gait of spinal cord injury (SCI) patients. Two kinds of linear actuator were developed by using direct current (d.c.) motors for assisting the knee and hip joint of a gait orthosis. They were mounted on the knee and hip joint of a commercial advanced reciprocating gait orthosis (ARGO), and a new two-degree-of-freedom externally powered gait orthosis was thus developed. The orthosis was assessed through inter-subject experiments on five male adult complete SCI patients. Owing to the short training period available for the assisted gait, simultaneous operation of both joint actuators was not conducted: either the knee actuation or the hip actuation was executed only. Thus, the knee actuator and the hip actuator were assessed with a T12 subject and with subjects for T5, T8, T11, and T12 respectively. The motions of the gaits, assisted by the linear actuators, were measured by a Vicon 370 system, and the general gait parameters and compensatory motions were evaluated. Results demonstrated that (a) all subjects could walk without falling, assisted either by the knee or the hip actuator; (b) both the knee and hip joint actuator increased the gait speed and the step length; (c) the knee flexion produced by the orthosis improved the dynamic cosmesis of walking; and (d) lateral compensatory motions as well as vertical ones tended to decrease when the hip joint was assisted, which could contribute to a reduction in walking effort.     

Determining the absolute efficiency of a delay line microchannel-plate detector using molecular dissociation

We present a method to measure the absolute detection efficiency of a delay-line microchannel-plate detector using the breakup of diatomic molecular ions. This method provides the absolute total detection efficiency, as well as the individual efficiency for each signal of the detector. The method is based on the fact that molecular breakup always yields two hits on the detector, but due to finite detection efficiency some of these events are recorded as single particles while others are detected in pairs. We demonstrate the method by evaluating the detection efficiency for both timing and position signals of a delay-line detector using laser-induced dissociation of molecular ions. In addition, the detection efficiency as a function of position has been determined by dividing the detector into sectors.     

Lead-free multilayer piezoelectric transformer

In this article, a multilayer piezoelectric transformer based on lead-free Mn-doped 0.94(Bi(12)Na(12))TiO(3)-0.06BaTiO(3) ceramics is presented. This piezoelectric transformer, with a multilayered construction in the thickness direction, is 8.3 mm long, 8.3 mm wide, and 2.3 mm thick. It operates in the second thickness extensional vibration mode. For a temperature rise of 20 degrees C, the transformer has an output power of >0.3 W. With a matching load resistance of 10 Omega, its maximum efficiency approaches 81.5%, and the maximum voltage gain is 0.14. It has potential to be used in low voltage power supply units such as low power adapter and other electronic circuits.     

Sensitivity of CFP/YFP and GFP/mCherry pairs to donor photobleaching on FRET determination by fluorescence lifetime imaging microscopy in living cells

Fluorescent protein-based FRET is a powerful method for visualizing protein-protein interactions and biochemical reactions in living cells. It can be difficult, however, to avoid photobleaching when observing fluorescent cells under the microscope, especially those expressing CFP. We compared the sensitivity of two protein-based FRET pairs to light-induced fluorescence changes in the donor, on FRET determination by fluorescence lifetime imaging microscopy (FLIM). Thanks to the very low excitation light levels of the time- and space-correlated single photon counting (TSCSPC) method, FLIM acquisitions were achieved without donor photobleaching. Here, we show that photobleaching of CFP by a mercury lamp under the microscope induced a decrease in the mean fluorescence lifetime, which interfered with FRET determination between CFP and YFP. Importantly, the range of light-induced variation of the mean fluorescence lifetime of CFP was not proportional to the decrease in the steady state fluorescence intensity and varied from cell to cell. The choice of the CFP/YFP pair therefore requires that the cells be observed and analyzed at very low light levels during the whole FRET experiment. In contrast, the GFP/mCherry pair provided an accurate FRET measurement by FLIM, even if some GFP photobleaching took place. We thus demonstrate that CFP can be an unreliable donor for FRET determination in living cells, due to its photosensitivity properties. We demonstrate that the GFP/mCherry pair is better suited for FRET measurement by FLIM in living cells than the CFP/YFP pair.