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We present the top–bottom effect (TBE) of a 5 µm thick amorphous specimen in a 3 MV ultrahigh voltage electron microscope (ultra-HVEM) and its influence on the quality of electron tomography (ET). The 40-nm gold particles on the top surface of the specimen tilted at different angles have been observed to be of poorer image quality than those on the symmetrical bottom surface obtained by tilting and turning over the specimen. The point spread function of the gold-particle images was calculated using the increment Wiener filter and the image-quality variation was then evaluated. The TBE is shown to become more remarkable with the increase of the effective thickness of the tilted specimen or the decrease of the magnification of the ultra-HVEM. The ET simulation indicates that the TBE may generate ∼4% distortion in the radial direction of a reconstructed sphere model. [ABSTRACT FROM AUTHOR]

Net photosynthetic rates (Pns) in leaves were compared between rice plants grown in ambient air control and free-air CO2 enrichment (FACE, about 200 µmol mol–1 above ambient) treatment rings. When measured at the same CO2 concentration, the Pn of FACE leaves decreased significantly, indicating that photosynthetic acclimation to high CO2 occurs. Although stomatal conductance (Gs) in FACE leaves was markedly decreased, intercellular CO2 concentrations (Ci) were almost the same in FACE and ambient leaves, indicating that the photosynthetic acclimation is not caused by the decreased Gs. Furthermore, carboxylation efficiency and maximal Pn, both light and CO2-saturated Pn, were decreased in FACE leaves, as shown by the Pn–Ci curves. In addition, the soluble protein, Rubisco (ribulose-1,5-bisphosphate caboxylase/oxygenase), and its activase contents as well as the sucrose-phosphate synthase activity decreased significantly, while some soluble sugar, inorganic phosphate, chlorophyll and light-harvesting complex II (LHC II) contents increased in FACE leaves. It appears that the photosynthetic acclimation in rice leaves is related to both ribulose-1,5-bisphosphate (RuBP) carboxylation limitation and RuBP regeneration limitation. [ABSTRACT FROM AUTHOR]

In electron tomography (ET) based on a transmission electron microscope, the effective thickness of a specimen increases with the tilt angle and, therefore, the projection quality may deteriorate because of electron scattering. The information-missing region, however, can be reduced by broadening the specimen tilt range. To clarify the general influence of these effects on ET, the projection quality varying with the tilt angle was quantitatively evaluated for a 5-μm thick specimen observed with a 3 MV ultrahigh-voltage electron microscope. Simulations of three-dimensional reconstruction were then performed for different specimen thicknesses and tilt ranges. As a result, the ET accuracy was shown to decrease as the specimen thickness increased. However, an optimumspecimen-tilt range, at which ET could reach its highest accuracy, was found to exist and become small with the increase of the specimen thickness. The presented results are helpful for determining the specimen thickness limitation on the ET resolution and improving the ET fidelity of thick specimens. [ABSTRACT FROM AUTHOR]

A one-dimensional model is proposed for analysing static capacitance contrast (SCC) in scanning electron microscopy. For the large-scale integrated specimen covered by an insulating thin film, the imaging signal is calculated considering the redistribution of secondary electrons (SEs) and the charging process of the equivalent effective capacitance between the irradiation point and substrate. The calculated SCC as a function of the irradiation charge density is in good agreement with the experimental SCC. It is confirmed that the SCC arises from the redistribution of SEs and the difference in the effective capacitance of irradiation points under the condition of positive charging. [ABSTRACT FROM AUTHOR]

We clarify the scanning electron microscopic contrast mechanism for imaging a buried SiO2 trench microstructure with interface trapped charges by simulating both electron scattering and transport. Here, the interface trapped charges make the SiO2 film more negatively charged and increase excess holes in the space charge distribution of the electron scattering region. The generated positive surface electric field thus redistributes some emitted secondary electrons and results in the dark contrast. This contrast mechanism is validated by comparing with experiments, and it may also provide an interesting approach for imaging and detecting deep interface trapped charges in insulating films. [ABSTRACT FROM AUTHOR]

Electron transmission through tilted epoxy-resin film specimens with thicknesses of 1 and 5 μm has been measured under different accelerating voltages and diameters of the objective aperture in a 3 MV ultrahigh voltage electron microscope. Measurements show that the linearity between the logarithmic electron transmission and effective thickness of the tilted specimens holds valid only for some smaller effective thicknesses. However, the higher accelerating voltage leads to a wider linear attenuation range. The linear range can reach 7 μm for the 3 MeV incident electrons. Moreover, the measurements were analyzed with the electron-scattering theory. Actually, the high-energy electrons are capable of penetrating much thicker specimens, but many electrons elastically scattered at large angles cannot be collected as useful signals because of being blocked by the objective aperture in the mode of amplitude contrast. The need for further investigations was suggested to determine theoretically the electron transmission through a thick low-Z specimen and its exponential attenuation range. [ABSTRACT FROM AUTHOR]