Nanometric locking of the tight focus for optical microscopy and tip-enhanced microscopy.

We have successfully stabilized the tight focus onto the sample surface of an optical microscope within ±1.0 nm for a virtually unlimited time duration. The time-dependent thermal drift of the tight focus and the mechanical tilt of the sample surface were simultaneously sensed by a non-optical means based on a capacitive sensor and were compensated for in real-time. This non-optical scheme is promising for the suppression of background light sources for optical microscopy. The focus stabilization is crucial for microscopic measurement at an interface, particularly when scanning a large surface area, because there is always a certain amount of mechanical tilt of the sample substrate, which degrades the contrast of the image. When imaging nanoscopic materials such as carbon nanotubes or silicon nanowires, more stringent nanometric stabilization of the focus position relative to such samples is required, otherwise it is often difficult to interpret the results from the observations. Moreover, the smaller the sample volume is, the smaller the signal becomes, resulting in a long exposure time at each position. In this sense, long-term stability of the tight focus is essential for both microscopic large area scanning and nanosized sample scanning (high-resolution/large-area imaging). In addition, the recently developed tip-enhanced microscopy requires long-term stability of the relative position of the tip, sample and focus position. We were able to successfully demonstrate a stability improvement for tip-enhanced microscopy in the same manner. The stabilization of the tight focus enables us to perform long-term and robust measurements without any degradation of optical signal, resulting in the capability of true nanometric optical imaging with good reproducibility and high precision. The technique presented is a simple add-on for any kind of optical microscope.

Characteristics of Mandibular Canal Branches Related to Nociceptive Marker.

It is difficult to correlate the direction of mandibular canal branches (MCBs) with altered sensation in dental treatments. In contrast, calcitonin gene-related peptide (CGRP) is related to vasodilation, bone formation, and the interaction with the peripheral nervous system. Therefore, we investigated the detailed morphological characteristics of MCBs using cone-beam computed tomography (CBCT) and observation of the CGRP distribution around the MCB. The MCB measurements were evaluated using principal component analysis (PCA) to identify morphological correlations. A total of 168 sides of mandibles from 84 cadavers were analyzed in this study. Most of the MCBs were primarily in the direction of the clock model from X to XI in sagittal sections and XII to I in coronal sections of the mandible. The structure of the MCB was divided into the fine canal branch (60.4%, 223/369), partial branch (24.4%, 90/369), and no canal branch (15.2%, 56/369). PCA indicated that the measurement element with the MCB and its structures were correlated in contrast to tooth factors. Positive CGRP reactions were clearly observed in the no-canal branch group compared to other groups. These data provide useful suggestions for MCB dynamics and information for clinical dental treatment.

Simulations of vacuum laser acceleration: hidden errors from particle's initial positions.

Simulation of vacuum laser acceleration, because of its scheme's simplicity, attracts many people involved in. However, how to put the particle in the initial positions in the field has not been considered seriously in some such schemes. An inattentive choice of electron's initial conditions may lead to misleading results. Here we show that arbitrarily placing the particle within the laser field leads to an overestimation of its energy gain, and offer suggestions for selecting appropriate initial conditions.

Nanoscale patterning induced strain redistribution in ultrathin strained Si layers on oxide.

We present a comparative study of the influence of the thickness on the strain behavior upon nanoscale patterning of ultrathin strained Si layers directly on oxide. The strained layers were grown on a SiGe virtual substrate and transferred onto a SiO(2)/Si substrate using wafer bonding and hydrogen ion induced exfoliation. The post-patterning strain was evaluated using UV micro-Raman spectroscopy for thin (20 nm) and thick (60 nm) nanostructures with lateral dimensions in the range of 80-400 nm. We found that about 40-50% of the initial strain is maintained in the 20 nm thick nanostructures, whereas this fraction drops significantly to approximately 2-20% for the 60 nm thick ones. This phenomenon of free surface induced relaxation is described using detailed three-dimensional finite element simulations. The simulated strain 3D maps confirm the limited relaxation in thin nanostructures. This result has direct implications for the fabrication and manipulation of strained Si nanodevices.

Mass infection with Entamoeba histolytica in a Japanese institution for individuals with mental retardation: epidemiology and control measures.

After two cases of amoebic colitis were detected at an institution for the mentally retarded in the Yamagata prefecture of Japan, the prevalence and epidemiology of Entamoeba histolytica infection at the institution were investigated. When the 76 residents with mental retardation were checked by serology and stool examinations, 40 (53%) showed evidence of infection with E. histolytica (i.e. E. histolytica-specific antibodies in their serum, Entamoeba cysts in their stools, and/or E. histolytica-specific antigens in their stools). The cysts were all assumed to be those of E. histolytica since all nine of the 18 cyst-positive stool samples investigated using a PCR (that distinguishes E. histolytica from E. dispar) were found positive for this species. The E. histolytica found in the institution in Yamagata appears to have been brought into the institution, from a similar institution in Kanagawa prefecture, by a mentally retarded individual who relocated from Kanagawa to Yamagata. Isolates of E. histolytica recovered during an outbreak in the institution in Kanagawa appear genotypically identical to the genotyped isolates collected in the outbreak investigated in the present study. The 40 infected individuals in Yamagata were each treated for 10 days with metronidazole or diloxanide furoate. The residents and staff of the institution were encouraged to wash their hands more frequently and more thoroughly, and the staff were asked to clip residents' fingernails and to improve the cleanliness/sterilization of the surfaces in the institution that were most likely to be contaminated with E. histolytica (lavatories, handrails, doors, doorknobs, washrooms, clothing etc). In the last 5 years of follow-up since the instigation of these and other infection-control measures, and the last treatments, no cases of E. histolytica infection have been found in the institution. This encouraging result offers hope and guidance to those attempting to control outbreaks of E. histolytica infection in other institutions.

Phase control of a z-current-driven plasma column.

A dynamic mitigation is presented for sausage and kink instability growths of a z-current-driven magnetized plasma column. In this Rapid Communication we found that a wobbling motion of the z-current electron axis induces a phase-controlled perturbation, so that the growths of the sausage and kink instabilities are successfully and remarkably mitigated. In general, plasma instabilities emerge from perturbations, and the perturbation phase is normally unknown. However, if the perturbation phase is known or actively imposed by, for example, a designed driver wobbling behavior, the instability growth would be controlled and mitigated by a superimposition of the perturbations imposed. The results in this Rapid Communication demonstrate that the wobbling z-current electron beam would provide an improvement in the plasma column stability and uniformity.

Peculiar behavior of Si cluster ions in a high-energy-density solid Al plasma.

Peculiar behavior is found in a Si cluster ion, moving with a speed of ∼0.22c (c: speed of light) in a solid Al plasma in the context of cluster-ion beam driven inertial confinement fusion: The Si ion, moving closely behind the forward-moving Si ion for a distance of several Šin the cluster, feels the wake field generated by the forward Si. The acceleration interaction force on the rear Si ion by the forward-moving ion may balance with the deceleration backward force in the longitudinal-moving direction. The forward-moving Si ion would be decelerated, as an isolated ion is decelerated without correlation. However, the deceleration of the rear Si ion, moving behind closely, would be reduced significantly. The rear Si ion may catch up and overtake the forward-moving Si ion in the cluster during the Si cluster-ion interaction with the high-density Al plasma. This peculiar behavior appears when the ions are aligned well longitudinally. The wake field is confined around the Si ion in the forward and transverse directions for a distance smaller than the Si cluster interionic distance l_{c}. However, the tail of the wake field extends beyond l_{c} due to the Si ion high speed of ∼0.22c. Therefore, the peculiar behavior shown above appears only for the ions in one cluster aligned well longitudinally.

Non-uniformity smoothing of direct-driven fuel target implosion by phase control in heavy ion inertial fusion.

We have proposed a dynamic smoothing method based on a phase control to smooth plasma non-uniformities in perturbed plasma systems. In this paper, the dynamic smoothing method is applied to a spherical direct-driven fuel target implosion in heavy ion inertial confinement fusion. We found that the wobbling motion of each heavy ion beam (HIB) axis induces a phase-controlled HIBs energy deposition, and consequently the phase-controlled implosion acceleration is realized, so that the HIBs irradiation non-uniformity is successfully smoothed. HIB accelerators provide a well-established performance to oscillate a HIB axis at a high frequency. In inertial confinement fusion, a fuel implosion uniformity is essentially significant for achieving the DT fuel compression and for releasing the fusion energy, and the non-uniformity of the implosion acceleration should be less than a few %. The results in this paper demonstrate that the wobbling HIBs would provide an improvement in the fuel target implosion uniformity.

Femtosecond laser nano-ablation in fixed and non-fixed cultured cells.

To understand the onset and morphology of femtosecond laser submicron ablation in cells and to study physical evidence of intracellular laser irradiation, we used transmission electron microscopy (TEM). The use of partial fixation before laser irradiation provides for clear images of sub-micron intracellular laser ablation, and we observed clear evidence of bubble-type physical changes induced by femtosecond laser irradiation at pulse energies as low as 0.48 nJ in the nucleus and cytoplasm. By taking ultrathin sliced sections, we reconstructed the laser affected subcellular region, and found it to be comparable to the point spread function of the laser irradiation. Laser-induced bubbles were observed to be confined by the surrounding intracellular structure, and bubbles were only observed with the use of partial pre-fixation. Without partial pre-fixation, laser irradiation of the nucleus was found to produce observable aggregation of nanoscale electron dense material, while irradiation of cytosolic regions produced swollen mitochondria but residual local physical effects were not observed. This was attributed to the rapid collapse of bubbles and/or the diffusion of any observable physical effects from the irradiation site following the laser exposure.

A femtosecond laser pacemaker for heart muscle cells.

The intracellular effects of focused near-infrared femtosecond laser irradiation are shown to cause contraction in cultured neonatal rat cardiomyocytes. By periodic exposure to femtosecond laser pulse-trains, periodic contraction cycles in cardiomyocytes could be triggered, depleted, and synchronized with the laser periodicity. This was observed in isolated cells, and in small groups of cardiomyocytes with the laser acting as pacemaker for the entire group. A window for this effect was found to occur between 15 and 30 mW average power for an 80 fs, 82 MHz pulse train of 780 nm, using 8 ms exposures applied periodically at 1 to 2 Hz. At power levels below this power window, laser-induced cardiomyocyte contraction was not observed, while above this power window, cells typically responded by a high calcium elevation and contracted without subsequent relaxation. This laser-cell interaction allows the laser irradiation to act as a pacemaker, and can be used to trigger contraction in dormant cells as well as synchronize or destabilize contraction in spontaneously contracting cardiomyocytes. By increasing laser power above the window available for laser-cell synchronization, we also demonstrate the use of cardiomyocytes as optically-triggered actuators. To our knowledge, this is the first demonstration of remote optical control of cardiomyocytes without requiring exogenous photosensitive compounds.

Stress imagining of semiconductor surface by tip-enhanced Raman spectroscopy.

Tip-enhanced Raman imaging of strained silicon reveals the property of nanoscale stress imposed on the lattice. Our approach relies on the highly localized excitation provided by a metallized tip. Surface sensitive detections in nanoscale are realized by a reflection-mode configuration combined with 442-nm excitation and a silver-coated silicon nitride tip. The background signals from an underlying silicon germanium layer and a tip are well suppressed. The quantitative stress analysis is made on the basis of the Raman shift of the Si-Si phonon mode. We succeeded in visualizing the localized stress with a spatial resolution down to 25 nm whereas a conventional micro Raman technique provides only a uniform image because of the averaging effect within a diffraction-limited focused spot.

Improvement of energy-conversion efficiency from laser to proton beam in a laser-foil interaction.

Improvement of energy-conversion efficiency from laser to proton beam is demonstrated by particle simulations in a laser-foil interaction. When an intense short-pulse laser illuminates the thin-foil target, the foil electrons are accelerated around the target by the ponderomotive force. The hot electrons generate a strong electric field, which accelerates the foil protons, and the proton beam is generated. In this paper a multihole thin-foil target is proposed in order to increase the energy-conversion efficiency from laser to protons. The multiholes transpiercing the foil target help to enhance the laser-proton energy-conversion efficiency significantly. Particle-in-cell 2.5-dimensional ( x, y, vx, vy, vz) simulations present that the total laser-proton energy-conversion efficiency becomes 9.3% for the multihole target, though the energy-conversion efficiency is 1.5% for a plain thin-foil target. The maximum proton energy is 10.0 MeV for the multihole target and is 3.14 MeV for the plain target. The transpiercing multihole target serves as a new method to increase the energy-conversion efficiency from laser to ions.

Localization of heparin-binding EGF-like growth factor in the smooth muscle cells and macrophages of human atherosclerotic plaques.

Heparin-binding EGF-like growth factor (HB-EGF) is a potent chemoattractant and mitogen for smooth muscle cells (SMC) in culture. To elucidate whether HB-EGF is implicated in the pathogenesis of human atherosclerosis, we examined immunohistochemical localization of HB-EGF in human aortic walls and atherosclerotic plaques. The medial SMC of the aorta in babies and children synthesized HB-EGF protein, while the number of SMC producing HB-EGF was dramatically decreased in young and middle-aged adults. In atherosclerotic plaques, however, marked production of HB-EGF protein was detected in SMC and macrophages of the plaques. Furthermore, EGF receptors, to which HB-EGF is known to bind, were detected in plaque SMC. These data suggest that HB-EGF may be implicated in the migration and proliferation of SMC that occurs in the normal development of arterial walls, and in the formation of atherosclerotic plaques.

Vascular endothelial dysfunction resulting from L-arginine deficiency in a patient with lysinuric protein intolerance.

Although L-arginine is the only substrate for nitric oxide (NO) production, no studies have yet been reported on the effect of an L-arginine deficiency on vascular function in humans. Lysinuric protein intolerance (LPI) is a rare autosomal recessive defect of dibasic amino acid transport caused by mutations in the SLC7A7 gene, resulting in an L-arginine deficiency. Vascular endothelial function was examined in an LPI patient who was shown to be a compound heterozygote for two mutations in the gene (5.3-kbp Alu-mediated deletion, IVS3+1G-->A). The lumen diameter of the brachial artery was measured in this patient and in healthy controls at rest, during reactive hyperemia (endothelium-dependent vasodilation [EDV]), and after sublingual nitroglycerin administration (endothelium-independent vasodilation [EIV]) using ultrasonography. Both EDV and NO(x) concentrations were markedly reduced in the patient compared with those for the controls. They became normal after an L-arginine infusion. EIV was not significantly different between the patient and controls. Positron emission tomography of the heart and a treadmill test revealed ischemic changes in the patient, which were improved by the L-arginine infusion. Thus, in the LPI patient, L-arginine deficiency caused vascular endothelial dysfunction via a decrease in NO production.

The anisotropic nanomovement of azo-polymers.

Nanoscale polymer movement is induced by a tightly focused laser beam in an azo-polymer film just at the diffraction limit of light. The deformation pattern that is produced by photoisomerization of the azo dye is strongly dependent on the incident laser polarization and the longitudinal focus position of the laser beam along the optical axis. The anisotropic photo-fluidity of the polymer film and the optical gradient force played important roles in the light induced polymer movement. We also explored the limits of the size of the photo-induced deformation, and we found that the deformation depends on the laser intensity and the exposure time. The smallest deformation size achieved was 200 nm in full width of half maximum; a value which is nearly equal to the size of the diffraction limited laser spot.

Slow Ca(2+) wave stimulation using low repetition rate femtosecond pulsed irradiation.

We demonstrated stimulation of Ca(2+) in living cells by near-infrared laser pulses operated at sub-MHz repetition rates. HeLa cells were exposed to focused 780 nm femtosecond pulses, generated by a titanium-sapphire laser and adjusted by an electro-optical modulator. We found that the laser-induced Ca(2+) waves could be generated over three orders of magnitude in repetition rates, with required laser pulse energy varying by less than one order of magnitude. Ca(2+) wave speed and gradients were reduced with repetition rate, which allows the technique to be used to modulate the strength and speed of laser-induced effects. By lowering the repetition rate, we found that the laser-induced Ca(2+) release is partially mediated by reactive oxygen species (ROS). Inhibition of ROS was successful only at low repetition rates, with the implication that ROS scavengers may in general be depleted in experiments using high repetition rate laser irradiation.

Three distinct types of Ca(2+) waves in Langendorff-perfused rat heart revealed by real-time confocal microscopy.

Although Ca(2+) waves in cardiac myocytes are regarded as arrhythmogenic substrates, their properties in the heart in situ are poorly understood. On the hypothesis that Ca(2+) waves in the heart behave diversely and some of them influence the cardiac function, we analyzed their incidence, propagation velocity, and intercellular propagation at the subepicardial myocardium of fluo 3-loaded rat whole hearts using real-time laser scanning confocal microscopy. We classified Ca(2+) waves into 3 types. In intact regions showing homogeneous Ca(2+) transients under sinus rhythm (2 mmol/L [Ca(2+)](o)), Ca(2+) waves did not occur. Under quiescence, the waves occurred sporadically (3.8 waves. min(-1) x cell(-1)), with a velocity of 84 microm/s, a decline half-time (t(1/2)) of 0.16 seconds, and rare intercellular propagation (propagation ratio <0.06) (sporadic wave). In contrast, in presumably Ca(2+)-overloaded regions showing higher fluorescent intensity (113% versus the intact regions), Ca(2+) waves occurred at 28 waves x min(-1) x cell(-1) under quiescence with a higher velocity (116 microm/s), longer decline time (t(1/2) = 0.41 second), and occasional intercellular propagation (propagation ratio = 0.23) (Ca(2+)-overloaded wave). In regions with much higher fluorescent intensity (124% versus the intact region), Ca(2+) waves occurred with a high incidence (133 waves x min(-1) x cell(-1)) and little intercellular propagation (agonal wave). We conclude that the spatiotemporal properties of Ca(2+) waves in the heart are diverse and modulated by the Ca(2+)-loading state. The sporadic waves would not affect cardiac function, but prevalent Ca(2+)-overloaded and agonal waves may induce contractile failure and arrhythmias.

Increase in the active form of 3-hydroxy-3-methylglutaryl coenzyme A reductase in human hepatocellular carcinoma: possible mechanism for alteration of cholesterol biosynthesis.

3-Hydroxy-3-methylglutaryl coenzyme A reductase activity and the rate of sterol biosynthesis are positively correlated with DNA synthesis and proliferation of mammalian cells. The total (active plus latent) activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase and the activity of its active form in hepatocellular carcinoma (HCC) from seven patients were measured and compared with those in liver tissue from five control subjects. The activity of the active form in HCC was 61 +/- 21 (SD) pmol/min/mg microsomal protein, while it was only 17 +/- 9.8 pmol/min/mg protein in the liver tissue from the controls; the difference was significant (P less than 0.005). The total activity of the reductase was also higher in HCC although the difference was not significant. The microsomal contents of the enzyme protein also were not significantly different. The rate of cholesterol biosynthesis was 307 +/- 81 pmol/h/mg tissue in HCC and 79.6 +/- 52 in normal liver tissue, indicating a significant increase in the rate in HCC (P less than 0.001). Thus, enhanced synthesis of cholesterol in human HCC seems to result partly from an increase in the active form of the reductase.

Elevated levels of transforming growth factor beta messenger RNA and its polypeptide in human hepatocellular carcinoma.

Malignant cells in culture express elevated levels of transforming growth factor beta 1 (TGF-beta 1) mRNA and secrete an abundant amount of TGF-beta protein, but little is known about the production of TGF-beta in human malignant tissues in vivo. We estimated the levels of TGF-beta 1 mRNA expression by Northern hybridization and measured TGF-beta protein using a radioreceptor assay in tumor tissues surgically obtained from six patients with hepatocellular carcinoma (HCC). TGF-beta 1 mRNA was expressed at much higher levels in HCC tissues from all the cases compared with normal human liver, suggesting an association of the activated TGF-beta 1 gene transcription with hepatocarcinogenesis. The content of TGF-beta was 207 +/- 121 ng/g wet tissue in the HCC tissue, and it showed correlation with the level of TGF-beta 1 mRNA in the tissue (r = 0.69; P less than 0.05). An immunohistochemical study demonstrated that TGF-beta 1 staining could be observed in HCC cells. These observations suggest that human HCC strongly expresses TGF-beta 1 mRNA in vivo, leading to a high content of TGF-beta protein.

Light-tunable Fano resonance in metal-dielectric multilayer structures.

High-Q optical Fano resonances realized in a variety of plasmonic nanostructures and metamaterials are very much promising for the development of new potent photonic devices, such as optical sensors and switches. One of the key issues in the development is to establish ways to effectively modulate the Fano resonance by external perturbations. Dynamic tuning of the Fano resonance applying the mechanical stress and electric fields has already been demonstrated. Here, we demonstrate another way of tuning, i.e., photo-tuning of the Fano resonance. We use a simple metal-dielectric multilayer structure that exhibits a sharp Fano resonance originating from coupling between a surface plasmon polariton mode and a planar waveguide mode. Using a dielectric waveguide doped with azo dye molecules that undergo photoisomerization, we succeeded in shifting the Fano resonance thorough photo-modulation of the propagation constant of the waveguide mode. The present work demonstrates the feasibility of photo-tuning of the Fano resonance and opens a new avenue towards potential applications of the Fano resonance.