High-Mobility Epitaxial Graphene on Ge/Si(100) Substrates.

Graphene was shown to reveal intriguing properties of its relativistic two-dimensional electron gas; however, its implementation to microelectronic applications is missing to date. In this work, we present a comprehensive study of epitaxial graphene on technologically relevant and in a standard CMOS process achievable Ge(100) epilayers grown on Si(100) substrates. Crystalline graphene monolayer structures were grown by means of chemical vapor deposition (CVD). Using angle-resolved photoemission spectroscopy and in situ surface transport measurements, we demonstrate their metallic character both in momentum and real space. Despite numerous crystalline imperfections, e.g., grain boundaries and strong corrugation, as compared to epitaxial graphene on SiC(0001), charge carrier mobilities of 1 × 104 cm2/Vs were obtained at room temperature, which is a result of the quasi-charge neutrality within the graphene monolayers on germanium and not dependent on the presence of an interface oxide. The interface roughness due to the facet structure of the Ge(100) epilayer, formed during the CVD growth of graphene, can be reduced via subsequent in situ annealing up to 850 °C coming along with an increase in the mobility by 30%. The formation of a Ge(100)-(2 × 1) structure demonstrates the weak interaction and effective delamination of graphene from the Ge/Si(100) substrate.

Minimum Resistance Anisotropy of Epitaxial Graphene on SiC.

We report on electronic transport measurements in rotational square probe configuration in combination with scanning tunneling potentiometry of epitaxial graphene monolayers which were fabricated by polymer-assisted sublimation growth on SiC substrates. The absence of bilayer graphene on the ultralow step edges of below 0.75 nm scrutinized by atomic force microscopy and scanning tunneling microscopy result in a not yet observed resistance isotropy of graphene on 4H- and 6H-SiC(0001) substrates as low as 2%. We combine microscopic electronic properties with nanoscale transport experiments and thereby disentangle the underlying microscopic scattering mechanism to explain the remaining resistance anisotropy. Eventually, this can be entirely attributed to the resistance and the number of substrate steps which induce local scattering. Thereby, our data represent the ultimate limit for resistance isotropy of epitaxial graphene on SiC for the given miscut of the substrate.

A versatile implementation of pulsed optical excitation in scanning tunneling microscopy.

We present a combination of pulsed optical excitation and scanning tunneling microscopy with a highly flexible pulse generation method. A high frequency arbitrary wave generator drives a gigahertz electro-optical modulator, which processes a continuous-wave laser beam of a low-noise laser diode into the desired wave shape. For pump-probe excitation we generate optical pulse series in an all-electronic way. Thereby we can easily adapt parameters like pulse amplitude, width, or repetition cycle to the demands of the experiment. This setup is used to study different dynamic processes at the GaAs(110) surface. Separating thermally induced effects from electrically induced effects allows us to quantify the thermal contribution of the optical excitation in STM experiments. Time-resolved decay spectra of the photo-generated electron-hole pairs demonstrate the nanoscale spatial resolution.

Local density of states at metal-semiconductor interfaces: an atomic scale study.

We investigate low temperature grown, abrupt, epitaxial, nonintermixed, defect-free n-type and p-type Fe/GaAs(110) interfaces by cross-sectional scanning tunneling microscopy and spectroscopy with atomic resolution. The probed local density of states shows that a model of the ideal metal-semiconductor interface requires a combination of metal-induced gap states and bond polarization at the interface which is nicely corroborated by density functional calculations. A three-dimensional finite element model of the space charge region yields a precise value for the Schottky barrier height.

Mapping itinerant electrons around Kondo impurities.

We investigate single Fe and Co atoms buried below a Cu(100) surface using low temperature scanning tunneling spectroscopy. By mapping the local density of states of the itinerant electrons at the surface, the Kondo resonance near the Fermi energy is analyzed. Probing bulk impurities in this well-defined scattering geometry allows separating the physics of the Kondo system and the measuring process. The line shape of the Kondo signature shows an oscillatory behavior as a function of depth of the impurity as well as a function of lateral distance. The oscillation period along the different directions reveals that the spectral function of the itinerant electrons is anisotropic.

Bistable charge configuration of donor systems near the GaAs(110) surfaces.

In gated semiconductor devices, the space charge layer that is located under the gate electrode acts as the functional element. With increasing gate voltage, the microscopic process forming this space charge layer involves the subsequent ionization or electron capture of individual dopants within the semiconductor. In this Letter, a scanning tunneling microscope tip is used as a movable gate above the (110) surface of n-doped GaAs. We study the build-up process of the space charge region considering donors and visualize the charge states of individual and multi donor systems. The charge configuration of single donors is determined by the position of the tip and the applied gate voltage. In contrast, a two donor system with interdonor distances smaller than 10 nm shows a more complex behavior. The electrostatic interaction between the donors in combination with the modification of their electronic properties close to the surface results in ionization gaps and bistable charge switching behavior.

A versatile high resolution scanning tunneling potentiometry implementation.

We have developed a new scanning tunneling potentiometry technique which can-with only minor changes of the electronic setup-be easily added to any standard scanning tunneling microscope (STM). This extension can be combined with common STM techniques such as constant current imaging or scanning tunneling spectroscopy. It is capable of performing measurements of the electrochemical potential with microvolt resolution. Two examples demonstrate the versatile application. First of all, we have determined local variations of the electrochemical potential due to charge transport of biased samples down to angstrom length scales. Second, with tip and sample at different temperatures we investigated the locally varying thermovoltage occurring at the tunneling junction. Aside from its use in determining the chemical identity of substances at the sample surface our method provides a controlled way to eliminate the influence of laterally varying thermovoltages on low-bias constant current topographies.

Enhanced donor binding energy close to a semiconductor surface.

We measured the ionization threshold voltage of individual impurities close to a semiconductor-vacuum interface, where we use the STM tip to ionize individual donors. We observe a reversed order of ionization with depth below the surface, which proves that the binding energy is enhanced towards the surface. This is in contrast to the predicted reduction for a Coulombic impurity in the effective mass approach. We can estimate the binding energy from the ionization threshold and show experimentally that in the case of silicon doped gallium arsenide the binding energy gradually increases over the last 1.2 nm below the (110) surface.

Electronic transport on the nanoscale: ballistic transmission and Ohm's law.

If a current of electrons flows through a normal conductor (in contrast to a superconductor), it is impeded by local scattering at defects as well as phonon scattering. Both effects contribute to the voltage drop observed for a macroscopic complex system as described by Ohm's law. Although this concept is well established, it has not yet been measured around individual defects on the atomic scale. We have measured the voltage drop at a monatomic step in real space by restricting the current to a surface layer. For the Si(111)-( [see text]3 x [see text]3)-Ag surface a monotonous transition with a width below 1 nm was found. A numerical analysis of the data maps the current flow through the complex network and the interplay between defect-free terraces and monatomic steps.

Controlled charge switching on a single donor with a scanning tunneling microscope.

The charge state of individually addressable impurities in semiconductor material was manipulated with a scanning tunneling microscope. The manipulation was fully controlled by the position of the tip and the voltage applied between tip and sample. The experiments were performed at low temperature on the (110) surface of silicon doped GaAs. Silicon donors up to 1 nm below the surface can be reversibly switched between their neutral and ionized state by the local potential induced by the tip. By using ultrasharp tips, the switching process occurs close enough to the impurity to be observed as a sharp circular feature surrounding the donor. By utilizing the controlled manipulation, we were able to map the Coulomb potential of a single donor at the semiconductor-vacuum interface.

Probing semiconductor gap states with resonant tunneling.

Tunneling transport through the depletion layer under a GaAs {110} surface is studied with a low temperature scanning tunneling microscope (STM). The observed negative differential conductivity is due to a resonant enhancement of the tunneling probability through the depletion layer mediated by individual shallow acceptors. The STM experiment probes, for appropriate bias voltages, evanescent states in the GaAs band gap. Energetically and spatially resolved spectra show that the pronounced anisotropic contrast pattern of shallow acceptors occurs exclusively for this specific transport channel. Our findings suggest that the complex band structure causes the observed anisotropies connected with the zinc blende symmetry.

Origin of Schottky barriers in gold contacts on GaAs110.

Gold contacts on n-type GaAs(110) have been investigated using scanning tunneling microscopy and spectroscopy in cross-sectional configuration. In spatially resolved current voltage spectroscopy the Schottky barrier potential is visible. We find signatures of delocalized gap states at the interface decaying into the semiconductor and observe a defect density at the interface below 3 x 10(13) cm(-2). Both findings support that the Fermi level pinning at the Au/GaAs(110) interface is dominated by metal-induced gap states.

Comparative activity of cisplatin, ifosfamide, doxorubicin, carboplatin, and etoposide in heterotransplanted hepatoblastoma.

BACKGROUND: Hepatoblastoma is the most common primary malignant liver tumor affecting infants and young children. Recent clinical experience with advanced hepatoblastoma shows that a reliable in vivo model to study the tumor's response to drugs is needed urgently. METHODS: Hepatoblastoma cell suspensions from three children were transplanted subcutaneously into NMRI nude mice (nu/nu). One of the primary tumors was a embryonal multifocal hepatoblastoma, whereas the other tumors were embryonal/fetal hepatoblastomas localized to one liver lobe. The xenograft tumors resembled their original tumors histologically and produced high levels of alpha-fetoprotein. The mice who received the tumors were given ifosfamide, cisplatin, doxorubicin, carboplatin, and etoposide as single agents. Thereafter, the tumor growth rate and alpha-fetoprotein levels in the animal sera were measured before and after chemotherapy and compared with the control group. After chemotherapy, the tumors were studied by conventional histology. RESULTS: The tumors in the nude mice derived from the multifocal hepatoblastoma reacted minimally against four of the five cytotoxic agents, whereas cisplatin reduced the tumor volume significantly. There was a marked reduction in tumor volume in the other tumors after application of cisplatin and doxorubicin, respectively. The tumors displayed a moderate reduction in size after treatment with ifosfamide, etoposide, and carboplatin. The responses to the different cytostatic agents also corresponded with serum alpha-fetoprotein levels and mitotic rates in the tumor cells. CONCLUSIONS: To the authors' knowledge, this is the first time an in vivo model for analyzing the effects of chemotherapy on hepatoblastoma has been established. The animal model may be suited for the evaluation of new drugs for the treatment of hepatoblastoma and for the investigation of multidrug resistance mechanisms in hepatoblastoma.

Altered expression of resistance associated genes in hepatoblastoma xenografts incorporated into mice following treatment with adriamycin or cisplatin.

To study the development of drug resistance in childhood hepatoblastoma (HB) using a model closely related to in vivo conditions, we cultivated HB xenografts from 3 patients into mice and treated these tumors with either adriamycin (ADR) or cisplatin (CIS). Determination of the relative expression levels of various resistance associated genes by a cDNA-PCR approach showed: a) Significantly enhanced MDR1 gene expression levels after treatment with ADR in each case. b) Significantly enhanced glutathione S-transferase mu (GST mu) gene expression levels after treatment with CIS in 2/3 xenografts. c) Significantly decreased levels of topoisomerase IIa (TOPO IIa) in tumors of the same two patients after treatment with either ADR or CIS. These findings give evidence that the MDR1-, GST mu- and TOPO IIa-gene products may contribute to drug resistance in HB.

A novel cytoplasmic dual specificity protein tyrosine phosphatase implicated in muscle and neuronal differentiation.

Dual specificity protein tyrosine phosphatases (dsPTPs) are a subfamily of protein tyrosine phosphatases implicated in the regulation of mitogen-activated protein kinase (MAPK). In addition to hydrolyzing phosphotyrosine, dsPTPs can hydrolyze phosphoserine/threonine-containing substrates and have been shown to dephosphorylate activated MAPK. We have identified a novel dsPTP, rVH6, from rat hippocampus. rVH6 contains the conserved dsPTP active site sequence, VXVHCX2GX2RSX5AY(L/I)M, and exhibits phosphatase activity against activated MAPK. In PC12 cells, rVH6 mRNA is induced during nerve growth factor-mediated differentiation but not during insulin or epidermal growth factor mitogenic stimulation. In MM14 muscle cells, rVH6 mRNA is highly expressed in proliferating cells and declines rapidly during differentiation. rVH6 expression correlates with the inability of fibroblast growth factor to stimulate MAPK activity in proliferating but not in differentiating MM14 cells. rVH6 protein localizes to the cytoplasm and is the first dsPTP to be localized outside the nucleus. This novel subcellular localization may expose rVH6 to potential substrates that differ from nuclear dsPTPs substrates.

Differential activation of mitogen-activated protein kinase in response to basic fibroblast growth factor in skeletal muscle cells.

In the MM14 mouse myoblast cell line, fibroblast growth factor (FGF) stimulates proliferation and represses differentiation. However, the intracellular signaling pathways used by FGF to affect these cellular processes are unknown. The predominant FGF receptor present on MM14 cells, FGFR1, is a receptor tyrosine kinase capable of activating the mitogen-activated protein kinase (MAPK) cascade in fibroblast and neuronal cell lines. To determine whether the FGF signal is mediated via the MAPK cascade in myoblasts, MM14 cells were stimulated with basic FGF (bFGF) and activities of the various kinases were measured. After withdrawal from serum and bFGF for 3 hr, bFGF stimulated MAPK kinase (MAPKK) activity, but MAPK and S6 peptide kinase activities were not detected. In contrast, when serum and bFGF were withdrawn for 10 hr, the activities of MAPKK, MAPK, and S6 peptide kinase were all stimulated by bFGF treatment. The inability of bFGF to stimulate MAPK after 3 hr of withdrawal may be due, in part, to the presence of a MAPK phosphatase activity that was detected in MM14 cell extracts. This dephosphorylating activity diminishes during commitment to terminal differentiation and is inhibited by sodium orthovanadate. Thus, the ability of bFGF to stimulate MAPK in MM14 cells is correlated with the loss of a MAPK phosphatase activity. These results show that although bFGF activates MAPKK in proliferating myoblasts, the mitogenic signal does not progress to the downstream kinases, providing a physiological example of an uncoupling of the MAPK cascade.

Remodeling of myofibrils: subcellular distribution of myosin heavy chain mRNA and protein.

Myofibril remodeling occurs during the normal life, in growth and development, and in the special case when isoform switching occurs. In this review we concentrate on the ultrastructural aspects of how myosin is incorporated into the A-band. Anatomic methods of study are emphasized that include isotope and immunochemical labeling as well as in situ hybridization. We conclude that the mechanism of remodeling is one of continual orderly exchange between a monomeric myosin pool and the thick filament. Myosin mRNA distribution is intermyofibrillar and nonsarcomeric, which suggests that newly translated myosin is released before diffusion and insertion in the A-band of the myofibrillar lattice.

Ultrastructural distribution of myosin heavy chain mRNA in cardiac tissue: a comparison of frozen and LR White embedment.

Electron microscopy (EM) in situ hybridization provides the higher resolution necessary to determine the spatial relationship between a specific mRNA and the organelle containing the protein encoded by that message. EM in situ hybridization was used to determine the subcellular myosin heavy chain (MHC) mRNA distribution with respect to the myofibril in normal cardiac tissue. Sections of frozen or acrylic-embedded tissue were compared for ultrastructural integrity and content of endogenous mRNA. Papillary muscles dissected from hearts of normal rabbits were aldehyde-fixed and either frozen or embedded in LR White. EM in situ hybridization with no riboprobe, vector sequence, same-sense, and anti-sense biotinylated riboprobes was detected by indirect immunocytochemistry. Labeling density using an antisense probe was highest over the intermyofibrillar space, with an average signal five times that of background. Background labeling by nonspecific sense probe was consistently low but not random, also having the highest density of gold clusters over the intermyofibrillar space. Ultracryomicrotomy yielded a higher absolute number of gold clusters, but sections were fragmented and disrupted striated muscle morphology. LR White embedment maintained ultrastructural integrity but gave a lower absolute signal. Fortunately, MHC mRNA is an abundant message and can tolerate the decreased sensitivity of LR White.

Distribution of myosin heavy chain mRNA in normal and hyperthyroid heart.

Hyperthyroid treatment produces rapid cardiac cell hypertrophy with all subcellular components increasing in an orderly manner. We compare normal and hyperthyroid tissue in order to relate changes in distribution of myosin mRNA during rapid assembly of myofibrils. At the light microscopic level, in situ hybridization of the ventricular cells shows myosin heavy chain mRNA to be distributed in a spoke-like pattern radiating from the nucleus. Electron microscopy provides the higher resolution necessary to determine mRNA distribution with respect to adjacent sarcomeric and cytoskeletal structures. Papillary muscles were removed from hyperthyroid and normal rabbits, aldehyde fixed, and embedded in LR white. Biotinated riboprobe transcribed from 0.5 kb in the coding region of terminal portion of the rod of alpha-myosin was hybridized and detected by immunocytochemical methods using 5 nm immunoglobulin G gold conjugates. Electron microscopy in situ hybridization runs with same-sense and anti-sense riboprobes were processed and ten micrographs randomly taken from each. Specific cytoplasmic densities of myosin mRNA were calculated by counting clusters of five or more gold particles over respective tissue components after subtraction of background counts. For both normal myocytes and hyperthyroid myocytes, the density of myosin mRNA was about 15 times higher in the cytoskeletal-rich inter-myofibrillar space than in the myofibrils. About half of the myosin mRNA in this inter-myofibrillar region is found within 10 nm of the peripheral filament, but no excess sarcomeric accumulation was seen beside the A-Band. It appears that most of the myosin is translated from mRNA within the inter-myofibrillar space along the entire length of the myofibril periphery. The emerging myosin heavy chain is not directly anchored to the thick filaments in either normal or rapidly growing cardiac cells.

Individual rabbit cardiac myocytes have different thresholds for alpha myosin heavy chain regulation by thyroid hormone.

Myocytes in adult rabbit ventricle express and alpha and a beta form of myosin heavy chain (MHC). The alpha-MHC distribution detected with indirect immunofluorescence has been found in different proportions in adjacent myocytes producing a mosaic staining pattern. The basis for cell-specific expression of the alpha-MHC isoform is not known. Since thyroid hormone is a major regulator of myosin gene expression, we varied the plasma thyroid level and followed the alpha-MHC content within a population of myocytes. Ventricular myocytes were induced to become 100% beta-MHC by placing the rabbits on a 0.15% propylthiouracil diet for 70 days. L-triiodothyronine (LT3) over a dose range of 1 to 10 micrograms/kg/day was delivered by an osmotic minipump for 5 days, with actual serum levels confirmed by LT3 radioimmunoassay to be in the range of from 115 to 1,230 ng/dl. The amount of alpha-MHC that returned was estimated in randomly selected cells by measuring the relative intensity of the fluorescence-tagged secondary antibody. The normal mosaic pattern of alpha-MHC expression in the left ventricle returned with an LT3 dose of 2-5 micrograms/kg/day. The first myocytes to express alpha-MHC were in the subepicardium and did so at a LT3 serum level of 115 of ng/dl. All myocytes of the ventricular wall expressed alpha-MHC at serum levels above 1,230 ng/dl. These data are interpreted to show that the variation of myosin isoform content seen in the adult heart is indicative of heterogeneity of thyroid sensitivity, with the threshold for serum LT3 being between 115 and 370 ng/dl.