Enhanced Eshelby twist on thin Wurtzite InP nanowires and measurement of local crystal rotation.

We have performed a detailed study of the lattice distortions of InP wurtzite nanowires containing an axial screw dislocation. Eshelby predicted that this kind of system should show a crystal rotation due to the dislocation induced torque. We have measured the twisting rate and the dislocation Burgers vector on individual wires, revealing that nanowires with a 10-nm radius have a twist up to 100% larger than estimated from elasticity theory. The strain induced by the deformation has a Mexican-hat-like geometry, which may create a tube-like potential well for carriers.

Correction of EELS dispersion non-uniformities for improved chemical shift analysis.

We outline a simple routine to correct for non-uniformities in the energy dispersion of a post-column electron energy-loss spectrometer for use in scanning transmission electron microscopy. We directly measure the dispersion and its variations by sweeping a spectral feature across the full camera to produce a calibration that can be used to linearize datasets post-acquisition, without the need for reference materials. The improvements are illustrated using core excitation electron energy-loss spectroscopy (EELS) spectra collected from NiO and diamond samples. The calibration is rapid and will be of use in all EELS analysis, particularly in assessments of the chemical states of materials via the chemical shift of core-loss excitations.

Near-simultaneous dual energy range EELS spectrum imaging.

A system that allows the collection of the low loss spectrum and the core loss spectrum, covering different energy regions, at each pixel in a spectrum image is described. It makes use of a fast electrostatic shutter with control signals provided by the spectrum imaging software and synchronisation provided by the CCD camera controller. The system also allows simultaneous collection of the X-ray spectrum and the signals from the imaging detectors while allowing the use of the existing features of the spectrum imaging software including drift correction and sub-pixel scanning. The system allows acquisition of high-quality spectra from both the core and the low loss regions, allowing full processing of the EELS data. Examples are given to show the benefits, including deconvolution, absolute thickness mapping and determination of numbers of atoms per unit area and per unit volume. Possible further developments are considered.

Quantitative electron energy-loss spectroscopy (EELS) analyses of lead zirconate titanate.

Electron energy-loss spectroscopy (EELS) analyses have been performed on a sol-gel deposited lead zirconate titanate film, showing that EELS can be used for heavy as well as light element analysis. The elemental distributions within the sol-gel layers are profiled using the Pb N(6,7)-edges, Zr M-edges, Ti L-edges and O K-edge. A multiple linear least squares fitting procedure was used to extract the Zr signal which overlaps with the Pb signal. Excellent qualitative information has been obtained on the distribution of the four elements. The non-uniform and complementary distributions of Ti and Zr within each sol-gel deposited layer are observed. The metal:oxygen elemental ratios are quantified using experimental standards of PbTiO(3), PbZrO(3), ZrO(2) and TiO(2) to provide relevant cross-section ratios. The quantitative results obtained for Ti/O and Pb/O are very good but the Zr/O results are less accurate. Methods of further improving the results are discussed.

Prolactin delays hair regrowth in mice.

Mammalian hair growth is cyclic, with hair-producing follicles alternating between active (anagen) and quiescent (telogen) phases. The timing of hair cycles is advanced in prolactin receptor (PRLR) knockout mice, suggesting that prolactin has a role in regulating follicle cycling. In this study, the relationship between profiles of circulating prolactin and the first post-natal hair growth cycle was examined in female Balb/c mice. Prolactin was found to increase at 3 weeks of age, prior to the onset of anagen 1 week later. Expression of PRLR mRNA in skin increased fourfold during early anagen. This was followed by upregulation of prolactin mRNA, also expressed in the skin. Pharmacological suppression of pituitary prolactin advanced dorsal hair growth by 3.5 days. Normal hair cycling was restored by replacement with exogenous prolactin for 3 days. Increasing the duration of prolactin treatment further retarded entry into anagen. However, prolactin treatments, which began after follicles had entered anagen at 26 days of age, did not alter the subsequent progression of the hair cycle. Skin from PRLR-deficient mice grafted onto endocrine-normal hosts underwent more rapid hair cycling than comparable wild-type grafts, with reduced duration of the telogen phase. These experiments demonstrate that prolactin regulates the timing of hair growth cycles in mice via a direct effect on the skin, rather than solely via the modulation of other endocrine factors.

Prolactin signaling influences the timing mechanism of the hair follicle: analysis of hair growth cycles in prolactin receptor knockout mice.

Pituitary PRL regulates seasonal hair follicle growth cycles in many mammals. Here we present the first evidence implicating PRL in the nonseasonal, wave-like pelage replacement of laboratory mice. In this study we show that messenger RNA transcripts encoding the one long and two short forms of PRL receptor are present in the skin of adult and neonate mice. The receptor protein was immunolocalized to the hair follicle as well as the epidermis and sebaceous glands. Furthermore, PRL messenger RNA was detected within skin extracts, suggesting a possible autocrine/paracrine role. Analysis of the hair growth phenotype of PRL gene-disrupted mice (PRLR(-/-)) revealed a change in the timing of hair cycling events. Although no hair follicle development differences were noted in PRLR(-/-) neonates, observations of the second generation of hair growth revealed PRLR(-/-) mice molted earlier than wild types (PRLR(+/+)). The advance was greater in females (29 days) than in males (4 days), resulting in the elimination of the sexual dimorphism associated with murine hair replacement. Heterozygotes were intermediate between PRLR(-/-) and PRLR(+/+) mice in molt onset. Once initiated, the pattern and progression of the molt across the body were similar in all genotypes. Although all fiber types were present and appeared structurally normal, PRLR(-/-) mice had slightly longer and coarser hair than wild types. These findings demonstrate that PRL has an inhibitory effect on murine hair cycle events. The pituitary PRL regulation of hair follicle cycles observed in seasonally responsive mammals may be a result of pituitary PRL interacting with a local regulatory mechanism.

Inhibitory effect of increased photoperiod on wool follicle growth.

The relationships between circulating prolactin (PRL), wool follicle growth and daylength were investigated in 24 New Zealand Wiltshire ewes housed indoors from September 1989 to May 1991. Twelve control (C) ewes were maintained under natural photoperiod. Two other groups were held in short days (SD; 8 h light: 16 h darkness) commencing from the winter solstice (22 June 1990) for either three (group SD3, n = 7) or six (group SD6, n = 5) months before reversion to natural daylength. Skin was sampled at one- to four-week intervals for histological determination of percentages of growing primary and secondary follicles. Hourly blood samples over 24 h were collected via jugular cannulae from C sheep in March and July and then monthly from all animals until December 1990 for estimation of mean monthly PRL concentrations for each treatment group. Between autumn (March 1990) and winter (July) primary follicle activity (PFA) and secondary follicle activity (SFA) declined in C ewes (PFA: 97 to 43%, SFA: 100 to 57%). Follicle regrowth during July and August in eight C ewes preceded the initial rise in plasma PRL from the winter minimum (1.6 ng/ml). Across the three groups, four instances of decreased follicle activity were observed, closely following or concurrent with increases in plasma PRL concentrations. The resumption of spring growth in four C sheep was temporarily checked by falls in follicle activities during September and October as PRL concentrations began to increase (3.4 to 8.9 ng/ml). Follicle activity also declined in November and December in eight C sheep, coincident with the rapid rise in PRL to a seasonal maximum in late November (165.4 ng/ml). The increase in SD3 follicle activity over spring was not delayed by short days but during October, after release from treatment, PRL concentrations rose (1.8 to 12.0 ng/ml) and follicle activity declined (PFA: 65 to 38%, SFA: 68 to 43%). In SD6 ewes, PRL concentrations were suppressed (2.1 ng/ml) and relatively constant levels of follicle activity (PFA: 73%, SFA: 95%) were maintained throughout short-day treatment. Release of SD6 ewes into summer photoperiod in January 1991 temporarily interrupted follicle growth (PFA: 68 to 17%, SFA: 96 to 19%) and caused out-of-season shedding in March and April. Contemporary C follicle activities were high (PFA: 95%, SFA: 98%). These data suggest that natural and experimental increases in daylength have a short-term inhibitory effect on growing wool follicles which could be mediated through rising concentrations of plasma prolactin.

Regulation of prolactin receptor expression in ovine skin in relation to circulating prolactin and wool follicle growth status.

Seasonal patterns of hair growth are governed, at least in part, by levels of prolactin in circulation, and although receptors for prolactin (PRLR) have been demonstrated in hair follicles, little is known of their regulation in relation to follicular cycles. In this study, a photoperiod-generated increase in prolactin was used to induce a wool follicle cycle during which changes in PRLR expression in sheep skin were determined by ribonuclease protection assay and in situ hybridisation. mRNA for prolactin and both isoforms of PRLR were also detected in skin by reverse transcription and polymerase chain reaction. As circulating prolactin began to rise from low levels, PRLR mRNA in the skin initially fell. These changes immediately preceded the catagen (regressive) phase of the hair cycle. Further increase in prolactin resulted in up-regulation of PRLR during telogen (dormancy), particularly in the epithelial hair germ, to reach a peak during proanagen (reactivation). In anagen (when follicle growth was fully re-established), PRLR mRNA returned to levels similar to those observed before the induced cycle. Hence, this longer term rise and fall of PRLR expression followed that of plasma prolactin concentration with a lag of 12-14 days. PRLR mRNA was most abundant in the dermal papilla, outer root sheath, hair germ, skin glands and epidermis. Location of PRLR in the dermal papilla and outer root sheath indicates action of prolactin on the growth-controlling centres within wool follicles. These cycle-related patterns of PRLR expression suggest dynamic regulation of PRLR by prolactin, thereby modulating hormonal responsiveness of seasonally growing hair follicles.

Instrumentation, techniques, and applications of electron microscopy in the solid state physics group at Glasgow University.

This review discusses some of the work performed by the Solid State Physics Group at Glasgow University. A major aim of the group is to obtain quantitative information with high spatial resolution and to do this reliably requires a thorough understanding of both the instrumentation and the interactions between the electron beam and the specimen. Thus the first part of the review discusses those aspects of instrumentation and techniques that the group has considered in detail while the final part deals with applications which involve the study of a wide range of materials covering metallurgical, semiconductor, organic, and magnetic systems. In all these applications, the results from a range of techniques have been required to provide as complete a picture of the material as possible.

Improving the analysis of small precipitates in HSLA steels using a plasma cleaner and ELNES.

The change from producing high strength low alloy (HSLA) steel sheet by conventional thick slab casting to producing it by direct charged thin slab casting causes a major change in the evolution of the precipitation. A key area of interest is the composition of the sub-10nm precipitates used to produce dispersion hardening. Carbon extraction replicas are frequently used to study precipitates in steels and other metals. When used with annular dark field imaging, this technique gives high contrast images of the precipitates while the thin carbon film adds little background or additional characteristic signals to either electron energy loss spectra or energy dispersive X-ray spectra. The method has the additional major advantage of removing the ferromagnetic matrix when studying HSLA steels. However, when the precipitates contain carbon, the C K-edge is dominated by the contribution from the amorphous carbon film. A plasma cleaner can be used to thin this carbon film to approximately 0.5 nm or less and then the contribution from the carbon in the precipitate can be separated from that in the carbon film using the electron energy loss near edge structure. A similar approach can be taken to separate the oxygen content of the precipitate from that of oxides formed from low-level impurities in the amorphous carbon during the plasma thinning process. In most cases, the precipitate studied here contained little or no oxygen even for the smallest sizes examined (approximately 4 nm). The precipitates contain mainly nitrogen with little carbon. For some compositions, the precipitates are clearly sub-stoichiometric.

Fresnel fringes in STEM.

Experiments carried out with a field emmission STEM show several Fresnel fringes along specimen edges, similar to CTEM observation. Analysis of the data shows agreement with computer profiles for idealized edges and the influence of defocusing distance and collector aperture size is discussed. In very coherent detection conditions the number of visible fringes is limited maily by signal-to-noise ratio considerations. In order to observe the maximum number of fringes, a collector aperture must be chosen so that there is a compromise between the limits imposed by the coherence of the detection and the signal-to-noise ratio.

A fast beam switch for controlling the intensity in electron energy loss spectrometry.

The fast deflection system described in this paper is suitable for controlling the intensity reaching the detector of a magnetic sector electron spectrometer mounted below an analytical transmission electron microscope. Amongst other things, this allows the low loss region of the spectrum to be recorded with the same electron probe conditions used to record core losses, something that is essential for high spatial resolution studies. The plate assembly restricts the width of the electron distribution reaching the viewing screen to a strip approximately 17 mm wide in the direction approximately normal to the dispersion direction of the spectrometer. The resulting deflection has no detectable effect on the FWHM of the zero-loss peak for exposure times as short as 1 micros. At incident energies up to 300 keV, positioning the deflection plates in the 35 mm camera port above the viewing chamber allows voltages of < +/- 3 kV to deflect the electrons out of the spectrometer and beyond the edge of the annular detector. When the deflection is switched on, the electrons are deflected out of the spectrometer in << 40 ns and when the deflection is switched off, the electrons return to within 10 microm of the undeflected position within 100 ns. Thus, even at an exposure time of 30 micros, the smallest time likely to be used in practice with a GATAN 666 spectrometer, < 1% of the signal in the spectrum is from electrons whose scattering conditions differ from those in the undeflected position. The performance of the deflection system is such that it will also be suitable for use with the new and much faster GATAN ENFINA spectrometer system. At incident energies up to 200 keV and possibly up to 300 keV, deflection voltages of +/- 3 kV are sufficient to deflect the electrons off a 1 k x 1 k charge coupled device (CCD) camera placed below the photographic camera. Thus the deflection system can be used as a very fast, non-mechanical shutter for such a CCD camera.

ELNES investigations of the oxygen K-edge in spinels.

The results of a systematic study of the oxygen K-edge electron energy-loss spectroscopy (ELNES) from a series of aluminium- and chromium-containing spinels are presented. Extra fine structure in the region up to 10 eV above the edge onset is observed for the chromium-containing compounds and is assigned to transitions to states created by mixing of oxygen 2p and metal 3d orbitals. The experimental data has been simulated using the multiple scattering code, FEFF8. Good agreement was obtained in the case of magnesium aluminate, but relatively poor agreement was obtained in the case of the chromites. The possible fingerprints in the oxygen K-edge ELNES corresponding to a high degree of inversion the spinel structure and to a tetragonal distortion of the cubic structure are discussed.

Characterising the surface and interior chemistry of core-shell nanoparticles using scanning transmission electron microscopy.

A method for extracting core and shell spectra from core-shell particles with varying core to shell volume fractions is described. The method extracts the information from a single EELS spectrum image of the particle. The distribution of O and N was correctly reproduced for a nanoparticle with a TiN core and Ti-oxide shell. In addition, the O distribution from a nanoparticle with a Cu core and a Cu-oxide shell was obtained, and the extracted Cu L(2,3)-core and shell spectra showed the required change in EELS near edge fine structure. The extracted spectra can be used for multiple linear least squares fitting to the raw data in the spectrum image. The effect of certain approximations on numerical accuracy, such as treating the nanoparticle as a perfect sphere, as well as the intrinsic detection limits of the technique have also been explored. The technique is most suitable for qualitative, rather than quantitative, work.

A new analytical method for characterising the bonding environment at rough interfaces in high-k gate stacks using electron energy loss spectroscopy.

Determining the bonding environment at a rough interface, using for example the near-edge fine structure in electron energy loss spectroscopy (EELS), is problematic since the measurement contains information from the interface and surrounding matrix phase. Here we present a novel analytical method for determining the interfacial EELS difference spectrum (with respect to the matrix phase) from a rough interface of unknown geometry, which, unlike multiple linear least squares (MLLS) fitting, does not require the use of reference spectra from suitable standards. The method is based on analysing a series of EELS spectra with variable interface to matrix volume fraction and, as an example, is applied to a TiN/poly-Si interface containing oxygen in a HfO(2)-based, high-k dielectric gate stack. A silicon oxynitride layer was detected at the interface consistent with previous results based on MLLS fitting.

Probing magnetic order in EELS of chromite spinels using both multiple scattering (FEFF8.2) and DFT (WIEN2k).

The electron energy loss near edge structure on the O K-edge from chromite spinels contains fine structure from the hybridisation of the O p-orbitals and the Cr d-orbitals. Unlike the aluminates, a non-spin polarised calculation of this fine structure differs significantly from experimental observations. This is due to the large magnetic moment on the Cr. Calculations using simplified collinear ordering of the spins and the local spin density approximation give much improved agreement. A real space multiple scattering formalism and a reciprocal space density functional formalism give results in substantial agreement. In general, the actual spin arrangement of these chromites is not known since they are typically frustrated magnetic systems with ordering temperatures in the 10-20K range. The calculations are based on the hypothesis that dynamic short range order persists to room temperature over the time scale of the interaction with the fast electron. However, it is possible that the observed effects are due to the strong paramagnetism present at room temperatures but which it is not possible to simulate accurately at present.

The microanatomy, cell replication, and keratin gene expression of hair follicles during a photoperiod-induced growth cycle in sheep.

Exposure of New Zealand Wiltshire sheep to long days, following 24 weeks of short days, caused a synchronised out-of-season wool follicle growth cycle. Skin biopsies were collected at intervals between 3 and 30 days and follicles were examined by light microscopy in both transverse and longitudinal section to describe the regressive (catagen), resting (telogen) and regenerative (proanagen) stages of the induced growth cycle. Follicles were generally in the growing phase (anagen) during short day treatment but by day 20 after exposure to long day photoperiod. 16% of follicles were in late catagen. By day 52, all follicles were in various stages of catagen, telogen and proanagen. The progression through the cycle occurred more slowly, but was morphologically similar to follicle growth cycles reported in rodents and goats, induced by plucking or melatonin, respectively. Follicles in early catagen were rarely observed, possibly reflecting the brevity of this phase of the cycle. Late catagen follicles were distinguished by the presence of a brush end and an inner root sheath, the latter disappearing as follicles entered telogen. Immunocytochemistry of proliferating cell nuclear antigen provided evidence that mitotic activity in the follicle bulb ceased completely during the brief telogen phase. The simultaneous absence of type I intermediate filament keratin mRNA indicated that keratinocyte differentiation had also been interrupted. Cell proliferation was re-established in early proanagen prior to observable changes in the follicle microanatomy. The relatively synchronised follicle growth cycle induced by photoperiod manipulation represents a potentially useful model for the study of changes in follicle ultrastructure and the endocrine and biochemical regulation of seasonal hair growth patterns.

The use of coloured dye-extraction microspheres to measure blood flow in rabbit skeletal muscle: a validation study with special emphasis on repeated measurements.

We evaluated the use of coloured dye-extraction microspheres for measuring blood flow in the skeletal muscle of anaesthetized rabbits. Spheres were injected into the left atrium after sternotomy or lateral thoracotomy, or into the left ventricle via the left carotid artery. Both routes of administration produced adequate mixing of microspheres with the blood. In general, the values of flow obtained from spheres of different colour were well correlated. Resting flows were difficult to measure, owing to the low number of spheres and the correspondingly low absorption of the samples. Occlusion of the capillary bed by trapped spheres was less than 10% in both tibialis anterior and extensor digitorum longus muscles after injection of a total of 15 x 10(6) spheres. Injection via the left ventricle, a closed-chest procedure, gave a more stable preparation than injections via the left atrium. However, in both cases there was a progressive decline in hyperaemic muscle blood flow during a series of injections. These results indicate that it is possible to use this technique to measure skeletal muscle blood flow in the rabbit, but that it is difficult to exploit the advantages of sequential microsphere injection in this species.

Bandstructure approach to near edge structure.

We review the current state of the art in EELS fingerprinting by computer simulation, focusing on the bandstructure approach to the problem. Currently calculations are made using a one electron theory, but we describe in principle the way to go beyond this to include final state effects. We include these effects within the one electron framework using the Slater transition state formula and assess the errors involved. Two examples are then given which illustrate the use of the one electron approximation within density functional theory. Our approach is to combine predicted atomic structure with predicted electronic structure to assist in fingerprinting of complex crystal structures.

Induction of a fast-oxidative phenotype by chronic muscle stimulation: histochemical and metabolic studies.

Chronic electrical stimulation of skeletal muscle at 10 Hz induces fast-to-slow fiber type transformation. Does a lower aggregate amount of activity lead to a less complete transformation, or does it produce the same transformation over a longer time course? We examined this question by subjecting adult rabbit tibialis anterior and extensor digitorum longus muscles to continuous stimulation at 2.5 Hz for 2-12 wk. Most of the fibers acquired the histochemical and immunocytochemical characteristics of type 2A, not type 1, fibers. There was a corresponding rise in oxidative activity, but this was accompanied by a marked decline in anaerobic glycolysis. The activities of hexokinase and 3-oxoacid CoA-transferase stopped increasing after 2 wk, glutamate oxaloacetate transaminase after 4 wk, and beta-hydroxyacyl-CoA dehydrogenase after 6 wk of stimulation. Succinate dehydrogenase, citrate synthase, lactate dehydrogenase, and creatine phosphokinase continued to change up to 12 wk of stimulation. Changes in enzyme activity were not as rapid or as marked as those observed for stimulation at 10 Hz, and none showed the typical two-phase response of oxidative enzyme activities to stimulation at 10 Hz. The latter may therefore be dependent on induction of type 1 myosin isoforms.