Evaluation of TEM specimen quality prepared by focused ion beam using symmetry breaking index of convergent-beam electron diffraction.

Degradation of the crystalline quality of transmission electron microscopy specimens in silicon prepared with different conditions has been examined using convergent-beam electron diffraction (CBED). The specimens are prepared using focused ion beam (FIB) with different accelerating voltages, Ar-ion milling and crushing method. Symmetry breaking of CBED patterns was quantitatively evaluated by symmetry breaking index S, which has been previously reported. The degradation and inhomogeneity of the FIB specimen were suppressed by decreasing the accelerating voltages of the FIB fabrication in the final process.

Evaluation of accuracy in the determination of crystal structure factors using large-angle convergent-beam electron diffraction patterns.

The accuracy of electron density distribution analysis using large-angle convergent-beam electron diffraction (LACBED) patterns is evaluated for different convergence angles. An orbital ordered state of FeCr2O4 is used as an example of the analysis. Ideal orbital-ordered and non-ordered states are simulated by using orbital scattering factors. LACBED patterns calculated for the orbital-ordered state were used as hypothetical experimental data sets. Electron density distribution of the Fe 3d orbitals has been successfully reconstructed with a higher accuracy from LACBED patterns with convergence angles larger than 15.2 mrad, which is 4 times as large as that for conventional convergent-beam electron diffraction patterns. Excitation of particular Bloch waves with the aid of LACBED patterns has a key role in the accurate analysis of electron density distributions.

Structural-transition-driven antiferromagnetic to spin-glass transition in Cd-Mg-Tb 1/1 approximants.

The magnetic susceptibility of the 1/1 approximants to icosahedral quasicrystals in a series of Cd85-xMgxTb15 (x = 5, 10, 15, 20) alloys was investigated in detail. The occurrence of antiferromagnetic to spin-glass-like transition was noticed by increasing Mg. Transmission electron microscopy analysis evidenced a correlation between the magnetic transition and suppression of the monoclinic superlattice ordering with respect to the orientation of the Cd4 tetrahedron at T > 100 K. The possible origins of this phenomenon were discussed in detail. The occurrence of the antiferromagnetic to spin-glass-like magnetic transition is associated with the combination of chemical disorder due to a randomized substitution of Cd with Mg and the orientational disorder of the Cd4 tetrahedra.

Time-lapse imaging of Ca2+-induced swelling and permeability transition: Single mitochondrion study.

Mitochondrial functions are closely related to the membrane structure. Mitochondrial swelling, which is accompanied with dissipation of the crista structure and rupture of the outer membrane, have been observed as mitochondrial damage when mitochondria are under Ca2+-overload or oxidative stress. Although these phenomena have been well studied, the detailed behaviors of individual mitochondria upon swelling remain unknown. The aim of this study was to investigate the detailed behavior of mitochondrial volume upon addition of Ca2+. Here, we report for the first time, time-lapse measurements of single mitochondrion swelling and permeability transition induced by Ca2+ by optical microscopy. We added 220 µM Ca2+ to mitochondria, and found that 1) the swelling rate depended on the mitochondrion, 2) a small number of mitochondria showed step-like swelling, 3) cyclosporin A decreased the percentage of mitochondria that underwent swelling induced by Ca2+, but did not affect the amplitude of swelling, 4) permeability transition is necessary but not sufficient for Ca2+-induced swelling, 5) permeability transition is more sensitive to Ca2+ than swelling, 6) Ca2+ stimulated mitochondrial swelling after permeability transition. These results suggest that single mitochondrion measurement of swelling is a powerful tool for examining the regulation of mitochondrial structure.

Disordered skyrmion phase stabilized by magnetic frustration in a chiral magnet.

Magnetic skyrmions are vortex-like topological spin textures often observed to form a triangular-lattice skyrmion crystal in structurally chiral magnets with the Dzyaloshinskii-Moriya interaction. Recently, ß-Mn structure-type Co-Zn-Mn alloys were identified as a new class of chiral magnet to host such skyrmion crystal phases, while ß-Mn itself is known as hosting an elemental geometrically frustrated spin liquid. We report the intermediate composition system Co7Zn7Mn6 to be a unique host of two disconnected, thermal-equilibrium topological skyrmion phases; one is a conventional skyrmion crystal phase stabilized by thermal fluctuations and restricted to exist just below the magnetic transition temperature Tc, and the other is a novel three-dimensionally disordered skyrmion phase that is stable well below Tc. The stability of this new disordered skyrmion phase is due to a cooperative interplay between the chiral magnetism with the Dzyaloshinskii-Moriya interaction and the frustrated magnetism inherent to ß-Mn.

Current-induced dynamics of skyrmion strings.

Dynamics of string-like objects is an important issue in a broad range of physical systems, including vortex lines in superconductors, viscoelastic polymers, and superstrings in elementary particle physics. In noncentrosymmetric magnets, string forms of magnetic skyrmions are present as topological spin objects, and their current-induced dynamics has recently attracted intense interest. We show in the chiral magnet MnSi that the current-induced deformation dynamics of skyrmion strings results in transport response associated with the real-space Berry phase. Prominent nonlinear Hall signals emerge above the threshold current only in the skyrmion phase. We clarify the mechanism for these nonlinear Hall signals by adopting spin density wave picture to describe the moving skyrmion lattice; deformation of skyrmion strings occurs in an asymmetric manner due to the Dzyaloshinskii-Moriya interaction, which leads to the nonreciprocal nonlinear Hall response originating from an emergent electromagnetic field. This finding reveals the dynamical nature of string-like objects and consequent transport outcomes in noncentrosymmetric systems.

Current-Induced Nucleation and Annihilation of Magnetic Skyrmions at Room Temperature in a Chiral Magnet.

A magnetic skyrmion is a nanometer-scale magnetic vortex carrying an integer topological charge. Skyrmions show a promise for potential application in low-power-consumption and high-density memory devices. To promote their use in applications, it is attempted to control the existence of skyrmions using low electric currents at room temperature (RT). This study presents real-space observations for the current-induced formation and annihilation of a skyrmion lattice (SkL) as well as isolated skyrmions in a microdevice composed of a thin chiral magnet Co8 Zn9 Mn3 with a Curie temperature, TC ≈ 325 K, above RT. It is found that the critical current for the manipulation of Bloch-type skyrmions is on the order of 108 A m-2 , approximately three orders of magnitude lower than that needed for the creation and drive of ferromagnetic (FM) domain walls in thin FM films. The in situ real-space imaging also demonstrates the dynamical topological transition from a helical or conical structure to a SkL induced by the flow of DC current, thus paving the way for the electrical control of magnetic skyrmions.

Deformation of Topologically-Protected Supercooled Skyrmions in a Thin Plate of Chiral Magnet Co8Zn8Mn4.

Magnetic skyrmions in Co8Zn8Mn4 thin plates are observed to deform in a metastable state prepared in a magnetic-field-cooling process by way of the thermal-equilibrium skyrmion phase. In cooling, the disk-shape skyrmions change to bar- or L-shaped elongated form, whereas the skyrmion density is nearly conserved. The deformation of the skyrmions in the supercooled metastable phase is observed irrespective of the crystallographic orientation of the thin plate, whereas the elongation direction nearly aligns along the magnetic easy axis. It is proposed that the deformation should be induced by a large increase in magnetic modulation wavenumber when decreasing the temperature, whereas the topological protection of the skyrmions keeps the averaged skyrmion density constant.

Detection of swelling of single isolated mitochondrion with optical microscopy.

Volume regulation under osmotic loading is one of the most fundamental functions in cells and organelles. However, the effective method to detect volume changes of a single organelle has not been developed. Here, we present a novel technique for detecting volume changes of a single isolated mitochondrion in aqueous solution based on the transmittance of the light through the mitochondrion. We found that 70% and 21% of mitochondria swelled upon addition of a hypotonic solution and Ca(2+), respectively. These results show the potential of the present technique to detect the physiological volume changes of individual small organelles such as mitochondria.

Fibrosis and adipogenesis originate from a common mesenchymal progenitor in skeletal muscle.

Accumulation of adipocytes and collagen type-I-producing cells (fibrosis) is observed in muscular dystrophies. The origin of these cells had been largely unknown, but recently we identified mesenchymal progenitors positive for platelet-derived growth factor receptor alpha (PDGFRα) as the origin of adipocytes in skeletal muscle. However, the origin of muscle fibrosis remains largely unknown. In this study, clonal analyses show that PDGFRα(+) cells also differentiate into collagen type-I-producing cells. In fact, PDGFRα(+) cells accumulated in fibrotic areas of the diaphragm in the mdx mouse, a model of Duchenne muscular dystrophy. Furthermore, mRNA of fibrosis markers was expressed exclusively in the PDGFRα(+) cell fraction in the mdx diaphragm. Importantly, TGF-ß isoforms, known as potent profibrotic cytokines, induced expression of markers of fibrosis in PDGFRα(+) cells but not in myogenic cells. Transplantation studies revealed that fibrogenic PDGFRα(+) cells mainly derived from pre-existing PDGFRα(+) cells and that the contribution of PDGFRα(-) cells and circulating cells was limited. These results indicate that mesenchymal progenitors are the main origin of not only fat accumulation but also fibrosis in skeletal muscle.

Genetic background affects properties of satellite cells and mdx phenotypes.

Duchenne muscular dystrophy (DMD) is the most common lethal genetic disorder of children. The mdx (C57BL/10 background, C57BL/10-mdx) mouse is a widely used model of DMD, but the histopathological hallmarks of DMD, such as the smaller number of myofibers, accumulation of fat and fibrosis, and insufficient regeneration of myofibers, are not observed in adult C57BL/10-mdx except for in the diaphragm. In this study, we showed that DBA/2 mice exhibited decreased muscle weight, as well as lower myofiber numbers after repeated degeneration-regeneration cycles. Furthermore, the self-renewal efficiency of satellite cells of DBA/2 is lower than that of C57BL/6. Therefore, we produced a DBA/2-mdx strain by crossing DBA/2 and C57BL/10-mdx. The hind limb muscles of DBA/2-mdx mice exhibited lower muscle weight, fewer myofibers, and increased fat and fibrosis, in comparison with C57BL/10-mdx. Moreover, remarkable muscle weakness was observed in DBA/2-mdx. These results indicate that the DBA/2-mdx mouse is a more suitable model for DMD studies, and the efficient satellite cell self-renewal ability of C57BL/10-mdx might explain the difference in pathologies between humans and mice.

CD90-positive cells, an additional cell population, produce laminin alpha2 upon transplantation to dy(3k)/dy(3k) mice.

Laminin alpha2 is a component of skeletal and cardiac muscle basal lamina. A defect of the laminin alpha2 chain leads to severe congenital muscular dystrophy (MDC1A) in humans and dy/dy mice. Myogenic cells including myoblasts, myotubes, and myofibers in skeletal muscle are a possible source of the laminin alpha2 chain, and myogenic cells are thus proposed as a cell source for congenital muscular dystrophy therapy. However, we observed production of laminin alpha2 in non-myogenic cells of normal mice, and we could enrich these laminin alpha2-producing cells in CD90(+) cell fractions. Intriguingly, the number of CD90(+) cells increased dramatically during skeletal muscle regeneration in mice. This fraction did not include myogenic cells but exhibited a fibroblast-like phenotype. Moreover, these cells were resident in skeletal muscle, not derived from bone marrow. Finally, the production of laminin alpha2 in CD90(+) cells was not dependent on fusion with myogenic cells. Thus, CD90(+) cells are a newly identified additional cell fraction that increased during skeletal muscle regeneration in vivo and could be another cell source for therapy for lama2-deficient muscular dystrophy.

Stability of membrane potential in heart mitochondria: single mitochondrion imaging.

Mitochondrial membrane potential (delta psi(m)) plays an important role in cellular activity. Although delta psi(m) of intracellular mitochondria are relatively stable, the recent experiments with isolated mitochondria demonstrate that individual mitochondria show frequent fluctuations of delta psi(m). The current study is performed to investigate the factors that stabilize delta psi(m) in cells by observing delta psi(m) of individual isolated mitochondria with fluorescence microscopy. Here, we report that (1) the transient depolarizations are also induced for mitochondria in plasma membrane permeabilized cells, (2) almost all mitochondria isolated from porcine hearts show the transient depolarizations that is enhanced with the net efflux of protons from the matrix to the intermembrane space, and (3) ATP and ADP significantly inhibit the transient depolarizations by plural mechanisms. These results suggest that the suppression of acute alkalinization of the matrix together with the presence of ATP and ADP contributes to the stabilization of delta psi(m) in cells.