Contrast and resolution of scanning transmission electron microscope imaging modes

Image blurring due to delocalization of inelastic events was studied for scanning transmission electron microscopy (STEM) of unstained thin sections. The delocalization probability was obtained from the angular distribution of inelastic scattering, which was calculated from experimental electron loss spectra of organic samples. This probability was implemented in a Monte Carlo program to simulate the effects of multiple scattering and delocalization for STEM images collected by either the annular detector or the spectrometer, and images generated by a combination of these two signals. Depending on the illumination, the detector geometry and the energy-loss range selected for imaging the annular detector image is blurred by a non-negligible fraction of inelastically scattered electrons. Simultaneous acquisition of an inelastic image using a spectrometer allows the blurring to be reduced by calculation of either the ratio or the difference of the two darkfield signals. While inherent nonlinearities reduce the interpretability of ratio-contrast images, difference-contrast improves the visibility of details submerged in a diffuse background without introducing artifacts.     

The height of biomolecules measured with the atomic force microscope depends on electrostatic interactions

In biological applications of atomic force microscopy, the different surface properties of the biological sample and its support become apparent. Observed height differences between the biomolecule and its supporting surface are thus not only of structural origin, but also depend on the different sample-tip and support-tip interactions. This can result in negative or positive contributions to the measured height, effects that are described by the DLVO (Derjaguin, Landau, Verwey, Overbeek) theory. Experimental verification shows that the electrostatic interactions between tip and sample can strongly influence the result obtained. To overcome this problem, pH and electrolyte concentration of the buffer solution have to be adjusted to screen out electrostatic forces. Under these conditions, the tip comes into direct contact with the surface of support and biological system, even when low forces required to prevent sample deformation are applied. In this case, the measured height can be related to the thickness of the native biological structure. The observed height dependence of the macromolecules on electrolyte concentration makes it possible to estimate surface charge densities.     

Human erythrocyte band 3. Solubilization and reconstitution into two-dimensional crystals

Various polyoxyethylene alkylethers were used to extract integral proteins from human erythrocyte membranes. The solubilization power of these detergents and the oligomerization of solubilized band 3 were studied. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis revealed that short-chain detergents induced oligomers larger than the band 3 dimer. In contrast, after solubilization with long-chain detergents, the predominant band on SDS-containing gels was the monomeric band 3. Oligomerization in short-chain detergents occurred preferentially at room temperature whereas monomeric band 3 prevailed at 4 degrees C. Consistent with these results, negative stain electron microscopy of solubilized isolated band 3 showed larger complexes with short-chain detergents than with long-chain detergents. Cu2+/o-phenanthroline-induced crosslinking had no effect on size or shape of band 3 particles. Despite their rather heterogeneous dimensions, octylpolyoxyethylene-solubilized band 3 complexes assembled into two-dimensional trigonal lattices (a = b = 11 (+/- 0.5) nm) in the presence of dimyristoyl phosphatidylcholine. The unit cell exhibited a pronounced stain-filled region surrounded by three elongated morphological subunits. Each subunit most likely represents a band 3 dimer. Freeze-drying/metal-shadowing of reconstituted lattices revealed one large elevation per unit cell protruding from an otherwise smooth surface.     

Nonshivering thermogenesis during acute cold exposure in adult and aged C57BL/6J mice

In C57BL/6J adult and aged mice, housed at room temperature (22.5 +/- 1 degrees C), we measured O2 consumption and CO2 production and calculated metabolic heat production under conditions of anesthesia and myorelaxation during acute cold stimulation when body temperature was lowered 7.5 degrees C below control level. An independent group of mice was subjected to a three hour partial physical restraint at 6 degrees C and concentration of uncoupling protein (thermogenin) was measured in interscapular brown adipose tissue mitochondria at different times after cold exposure. Heat production under anesthesia and myorelaxation was about 57-66% lower than in nonanesthetized conditions, but increased significantly during cold stimulation in both age groups. Under anesthesia and myorelaxation before and during cold stimulation aged mice produced about 20% more heat than adult mice. Because in these experiments all sources of facultative thermogenesis, except nonshivering, were suppressed by anesthesia and myorelaxation, and because brown adipose tissue is the major source of nonshivering thermoproduction, we concluded that aged mice housed at room temperature have an increased thermogenesis in brown adipose tissue. This conclusion was also supported by the finding that the concentration of uncoupling protein measured in the mitochondria of brown adipose tissue after single cold exposure was significantly higher in aged than in adult mice. Therefore, we propose that the lower, cold-induced, heat production typically observed in nonanesthetized aged mice may reflect reduced thermogenic capacity of skeletal muscles. While aged mice have less brown adipose tissue than adult animals, the remaining brown adipose tissue may compensate by increasing the concentration of uncoupling protein.     

Characterization of a shielded transition to a dielectric waveguide

A shielded sub-mm/THz monolithic transition between a layered dielectric waveguide and a strip-ridge line is characterized using two different approaches: the integral equation-mode matching (IEMM) method and the finite difference time domain (FDTD) technique. While higher order modes are considered, a simple method for determining the transition's circuit model (i.e., two-port scattering matrix) from the IEMM results is implemented and the electrical performance is studied as frequency and conductor width are varied. The FDTD analysis gives further insight into the behavior of the transition in a very wide frequency range (0-540 GHz). The transition is found to be very efficient over a wide frequency band and a broad range of conductor widths     

Study of print-wire wear

This study demonstrates the dependence of wire printer wear on the abrasiveness of the impacted medium and the influence of ink in the ribbon. A predictive linear wear model is derived and its validity shown experimentally.