Hybrids of Magnetic Nanoparticles with Double‐Hydrophilic Core/Shell Cylindrical Polymer Brushes and Their Alignment in a Magnetic Field

The synthesis of double‐hydrophilic core/shell cylindrical polymer brushes (CPBs), their hybrids with magnetite nanoparticles, and the directed alignment of these magnetic hybrid cylinders by a magnetic field are demonstrated. Consecutive grafting from a polyinitiator poly(2‐(2‐bromoisobutyryloxy)ethyl methacrylate) (PBIEM) of tert‐butyl methacrylate (tBMA) and oligo(ethylene glycol) methacrylate (OEGMA) using atom‐transfer radical polymerization (ATRP) and further de‐protection yields core/shell CPBs with poly(methacrylic acid) (PMAA) as the core and POEGMA as the shell, which is evidenced by ¹H NMR, gel permeation chromatography (GPC), and dynamic and static light scattering (DLS and SLS). The resulting core/shell brush is well soluble in water and shows a pH responsiveness because of its weak polyelectrolyte core. Pearl‐necklace structures are observed by cryogenic transmission electron microscopy (cryo‐TEM) at pH 4, while at pH 7, these structures disappear owing to the ionization of the core. A similar morphology is also found for the polychelate of the core/shell CPBs with Fe³⁺ ions. Superparamagnetic magnetite nanoparticles have also been prepared and introduced into the core of the brushes. The hybrid material retains the superparamagnetic property of the magnetite nanoparticles, which is verified by superconducting quantum interference device (SQUID) magnetization measurements. Large‐scale alignment of the hybrid cylinders in relatively low magnetic fields (40–300 mT) can easily be performed when deposited on a surface. which is clearly revealed by the atomic force microscopy (AFM) and TEM measurements.     

Enzyme-histochemical demonstration of lymphatic vessels in the golden hamster periodontium: an attempt to reactivate the 5'-nucleotidase activity with Mg++ ion supply

Enzyme-histochemical demonstration of lymphatic vessels in the golden hamster periodontium was performed on cryostat sections using the 5'-nucleotidase (5'-Nase) staining method by light microscopy and backscattered electron imaging of scanning electron microscopy. The inhibition of the 5'-Nase activity by decalcification was cancelled by the Mg++ ion supply. The reaction products of 5'-Nase activity were produced on the lymphatic endothelial cells and the tubular structures of lymphatic vessels were seen as highlights by backscattered electron imaging. The invasion of 5'-Nase-positive lymphatic vessels into the alveolar bone from the periodontium was found in the present study.     

热氧化磁控溅射金属锌膜制备ZnO纳米棒

利用射频磁控溅射技术在Si(111)衬底上制备金属锌膜 ,在空气中退火热氧化合成了一维ZnO纳米棒。用X射线衍射 (XRD) ,扫描电子显微镜 (SEM) ,透射电子显微镜 (TEM)和光致发光谱 (PL)对样品进行了结构、形貌及光学特性分析。结果表明 :ZnO纳米棒为六方纤锌矿结构单晶相 ,直径在 30～ 6 0nm左右 ,其长度可达5～ 8μm左右。在 2 80nm波长光激发下 ,有很强的 372nm带边紫外光发射和较微弱的 5 16nm深能级绿光发射 ,说明合成的单晶ZnO纳米棒的质量较高     

拓宽激光扫描共聚焦显微技术运用的新探索

利用一些醛类固定液在固定作用中可产生荧光物质的特性，在荧光显微术中选择合适的激发波长和发射波长，使得一些未经荧光标记的生物组织结构也能在激光扫描共聚焦显微镜下成像，并可进行三维图像的构建，这不仅使激光扫描共聚焦显微技术的运用范围大为扩展，而且大大节约了研究费用和时间，提高了效益。此外，该技术还为组织学和细胞学的回顾性研究提供了一个有效手段，并将激光扫描共聚焦显微技术与普通光镜及电镜技术有机地结合起来，使组织和细胞的形态学研究得以全方位进行。     

In vivo subcellular ultrastructures recognized with Hilbert differential contrast transmission electron microscopy

We describe the first application of a novel electron microscopic technique to visualize subcellular structures in a near-living state. Rapidly frozen ice-embedded cells provide the most realistic images, as they are free from artefacts induced by sample preparation methods, such as chemical fixation, dehydration, staining and sectioning. The application of the conventional transmission electron microscope to ice-embedded cell imaging, however, has been limited by the low image contrast. The recently developed Hilbert differential contrast transmission electron microscope, which exhibits an unexpectedly high contrast, akin to the differential interference contrast in visible light microscopy, enabled us to clearly discern detailed subcellular structures in ice-embedded cyanobacterial cells.     

Evidence for glycogen structures associated with plasma membrane invaginations as visualized by freeze-substitution and the Thiery reaction in Saccharomyces cerevisiae

We have used a combination of freeze-substitution electron microscopy and specific reaction for polysaccharides to re-evaluate glycogen structures in Saccharomyces cerevisiae. We also used mutant cells devoid of glycogen to confirm the glycogenic nature of the structures described. Previously described cytoplasmic aggregates were confirmed as glycogen granules. Moreover, an original structure was discovered. This is a ring of glycogen surrounding a finger- or pleat-like plasma membrane invagination. This structure could be physiologically significant in terms of channelling glucose to or from glycogen reserves.     

Staining of intestinal goblet cells with ruthenium red in semithin sections.

For a correlative light and electron microscopy of intestinal goblet cells, postembedding staining with ruthenium red (RR) was performed in epoxy-embedded sections. Tissue blocks were fixed in buffered aldehyde and embedded in a mixture of Quetol 651, nonenyl succinic anhydride (NSA), methyl nadic anhydride (MNA), and DMP-30. Sections at 0.4-0.5 micron in thickness were mounted on grids and were treated with an aqueous solution of RR followed by osmium tetroxide, uranyl acetate and lead citrate. Postembedding staining of epoxy sections revealed the interaction between RR and anionic groups by both light and electron microscopy. Light and electron microscopic observation of identical sites in semithin sections was successful for the correlations of colored reaction with electron density.     

介孔ZnO微球的制备及其在染料敏化太阳能电池中的应用

制备了一种新型的染料敏化太阳电池的光阳极,该电极由溶剂合成的具有高比表面积和良好光散射特性的ZnO介孔微球组成。采用X射线衍射、扫描电子显微镜、能谱仪及N2吸附脱附等手段,分析了介孔ZnO微球的结构和形貌。所得介孔微球尺寸在亚微米范围,比表面积约为50m2.g-1。将ZnO介孔微球成功应用到染料敏化太阳电池中,当光阳极为3μm时,组成的原型器件的短路电流密度约为4.5mA.cm-2,开路电压约为602mV,转换效率可达1.28%。研究结果表明,ZnO介孔微球是一种优异的染料敏化太阳电池的光阳极材料。     

小麦颖果发育过程中韧皮部细胞的结构变化观察

利用光学显微镜、透射电镜和扫描电镜技术．对小麦颖果发育过程中韧皮部细胞的结构变化进行了系统观察。结果表明，开花后7d颖果还没有形成完全分化的维管束。开花后12d韧皮部可见明显的筛分子。每个维管束有大约40个左右的筛分子，它们呈半圆形分布于维管束外侧。开花后14～20d，筛分子内有较丰富的线粒体、P^-型质体和许多无定型丝状物，有些筛分子有珠光壁。筛分子有典型的伴胞，并形成SE／CC复合物结构。伴胞有稠密的胞质和核质。韧皮薄壁细胞与中间细胞间有丰富的胞间连丝；韧皮薄壁细胞间的胞间连丝呈区域集中分布。开花后24-29d，筛分子内线粒体数目减少，伴胞和韧皮薄壁细胞电子染色变浅，说明它们的养分运输能力下降。中间细胞的细胞质内仍有丰富的线粒体和其它细胞器，它们沿胞壁周缘分布；此时，中间细胞形成类似筛分子的结构．因此中间细胞是一种适应于养分运输的特化伴胞，在颖果灌浆后期起重要作用。开花后39d，韧皮部细胞衰老变形，此时颖果发育成熟。     

Ultrastructural disorder of actin mutant suggests uncoupling of actin-dependent pathway from microtubule-dependent pathway in budding yeast

Temperature-sensitive actin mutant of Saccharomyces cerevisiae act1-1 was studied at a permissive temperature of 23°C by light, fluorescent and electron microscopy to elucidate the roles of actin cytoskeleton in the cycling eukaryotic cells. Mutant cells that grew slowly at the permissive temperature showed aberrations in the cytoskeleton and cell cycle. Mutant cells contained aberrant 'faint actin cables,' that failed in directing of mitochondria, vacuoles and secretory vesicles to the bud and the stray vesicles delivered their content to the mother wall instead of the bud. Bud growth was delayed. Spindle pole bodies and cytoplasmic microtubules did not direct to the bud, and nucleus failed to migrate to the bud. Repeated nuclear divisions produced multinucleated cells, indicating continued cycling of actin mutant cells that failed in the morphogenetic checkpoint, the spindle position checkpoint and cytokinesis. Thus, a single actin mutation appears to indicate uncoupling in space and time of the 'actin cytoskeleton-dependent cytoplasmic pathway of bud development and organelle positioning and inheritance' from the 'microtubule-dependent nuclear division pathway' in a budding yeast cell cycle.     

Carotid body-like tissues around the rat posterior cricoarytenoid muscle

We examined the carotid body-like tissues around the posterior cricoarytenoid (PCA) muscle of the rat by light and electron microscopy. One branch after bifurcation of the inferior (recurrent) laryngeal nerve frequently formed a small ganglion at the lateral side of this muscle and sometimes contained paraganglion cells (granule-containing cells). In addition, encapsulated structures (paraganglia) enveloped by a few layer of capsular cells were often observed on and near the muscle. Moreover, granule-containing cells resembling the encapsulated paraganglion cells were found in clusters outside the small nerve. These clustered cells were incompletely surrounded by fibroblastic processes. In addition to synapses between adjacent cells, afferent and efferent synapses were distinguished between nerve endings and these cells, possibly receiving both afferent and efferent innervation. These findings suggest that the clustered granule-containing cells outside the small nerve in the proximity of the PCA muscle may function as chemoreceptor cells as well as the paraganglion cells within the nerve bundles and the encapsulated paraganglia.     

Transparent Poly(methyl methacrylate)/Single‐Walled Carbon Nanotube (PMMA/SWNT) Composite Films with Increased Dielectric Constants

Poly(methyl methacrylate)/single‐walled carbon nanotube (PMMA/SWNT) composites were prepared via in situ polymerization induced either by heat, ultraviolet (UV) light, or ionizing (gamma) radiation. The composites dissolved in methylene chloride and then cast into films exhibited enhanced transparency as compared with the melt‐blended composite material. UV/visible spectroscopy was used to quantitatively analyze the transparency of the composites. The dielectric constant (ε′) was measured via dielectric analysis (DEA) and correlated to the refractive‐index values using Maxwell's relationship. The dielectric constant increased in the composite samples as compared with the neat PMMA samples prepared by the same methods. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) provided images of the polymer–nanotube composites and single‐walled CNTs, respectively.     

3D Microperiodic Hydrogel Scaffolds for Robust Neuronal Cultures

Three-dimensional (3D) microperiodic scaffolds of poly(2-hydroxyethyl methacrylate) (pHEMA) have been fabricated by direct-write assembly of a photopolymerizable hydrogel ink. The ink is initially composed of physically entangled pHEMA chains dissolved in a solution of HEMA monomer, comonomer, photoinitiator and water. Upon printing 3D scaffolds of varying architecture, the ink filaments are exposed to UV light, where they are transformed into an interpenetrating hydrogel network of chemically cross-linked and physically entangled pHEMA chains. These 3D microperiodic scaffolds are rendered growth compliant for primary rat hippocampal neurons by absorption of polylysine. Neuronal cells thrive on these scaffolds, forming differentiated, intricately branched networks. Confocal laser scanning microscopy reveals that both cell distribution and extent of neuronal process alignment depend upon scaffold architecture. This work provides an important step forward in the creation of suitable platforms for in vitro study of sensitive cell types.     

Growth and characterization of AlxGa1−xAs Bragg reflectors by LP-MOCVD

Using a moderate-sized LP-MOCVD production reactor, we have demonstrated the growth of Bragg reflector structures that reflect nearly 100% of the light at the design wave-length of 850 nm. The designs studied consist of alternating layers of Al_0.10Ga_0.90As and Al_0.85Gao_0.15As, with up to 30 periods. Characterization data show the structures to be highly reproducible and uniform, with good agreement between data obtained by double-crystal x-ray rocking curve analysis, cross-sectional transmission electron microscopy, and optical-reflectance spectroscopy. GaAs solar cells utilizing Bragg reflectors have been fabricated and characterized.     

Bright White Scattering from Protein Spheres in Color Changing,Flexible Cuttlefish Skin

Throughout nature, elegant biophotonic structures have evolved into sophisticated arrangements of pigments and structural reflectors that manipulate light in the skin, cuticles, feathers and fur of animals. Not many spherical biophotonic structures are known and those described are often angle dependent or spectrally tuned. White light scattering by the flexible skin of cuttlefish (Sepia officinalis) is examined and how the unique structure and composition of leucophore cells serve as physiologically passive reflectors approximating the optical properties of a broadband Lambertian surface is investigated. Leucophores are cells that contain thousands of spherical microparticles called leucosomes that consist of sulfated glycoproteins or proteoglycans and reflectin. A leucophore containing ≈12 000 leucosome microspheres is characterized three‐dimensionally by electron microscopy and the average refractive index of individual leucosomes is measured by holographic microscopy to be 1.51 ± 0.02. Modeling of the ultrastructural data and spectral measurements with Lorenz‐Mie theory and Monte Carlo simulations suggest that leucophore whiteness is produced by incoherent scattering based upon a randomly ordered system. These soft, compliant, glycosylated proteinacious spheres may provide a template for bio‐inspired approaches to efficient light scattering in materials science and optical engineering.     

A useful method for observing intracellular structures of free and cultured cells by scanning electron microscopy

Scanning electron microscopy (SEM) using osmium-maceration methods has been used for analyzing the three-dimensional structure of cell organelles in tissue samples, but it has been quite difficult to observe free and cultured cells with this technique. The present study was performed to develop a method that can be applied to free and cultured cells for SEM studies of intracellular structures after osmium maceration. The method was also applied to light microscopy (LM) and to transmission electron microscopy (TEM). HeLa cells and human leukocytes were fixed with a mixture of 0.5% paraformaldehyde and 0.5% glutaraldehyde followed by an additional fixation with 1% osmium tetroxide. These cells were embedded in low-melting-point agarose. A temperature-responsive dish was also used for collection of cultured cells before embedding. For LM and TEM, the cell-embedded agarose was further embedded in epoxy resin, and semi- and ultrathin sections were examined conventionally. For SEM, the agarose was freeze-fractured in 50% dimethyl sulfoxide, processed for osmium maceration and observed in a high-resolution SEM. Low-melting-point agarose was useful as an embedding medium for SEM, because it was well preserved during prolonged osmication for SEM. Thus, the fine structure of cell organelles was clearly analyzed by SEM after osmium-maceration treatment. These SEM images could also be compared with those of LM and TEM of the agarose-embedded tissues.     

From electron microscopy to molecular cell biology, molecular genetics and structural biology: intracellular transport and kinesin superfamily proteins, KIFs: genes, structure, dynamics and functions

Cells transport and sort various proteins and lipids following synthesis as distinct types of membranous organelles and protein complexes to the correct destination at appropriate velocities. This intracellular transport is fundamental for cell morphogenesis, survival and functioning not only in highly polarized neurons but also in all types of cells in general. By developing quick-freeze electron microscopy (EM), new filamentous structures associated with cytoskeletons are uncovered. The characterization of chemical structures and functions of these new filamentous structures led us to discover kinesin superfamily molecular motors, KIFs. In this review, I discuss the identification of these new structures and characterization of their functions using molecular cell biology and molecular genetics. KIFs not only play significant roles by transporting various cargoes along microtubule rails, but also play unexpected fundamental roles on various important physiological processes such as learning and memory, brain wiring, development of central nervous system and peripheral nervous system, activity-dependent neuronal survival, development of early embryo, left-right determination of our body and tumourigenesis. Furthermore, by combining single-molecule biophysics with structural biology such as cryo-electrom microscopy and X-ray crystallography, atomic structures of KIF1A motor protein of almost all states during ATP hydrolysis have been determined and a common mechanism of motility has been proposed. Thus, this type of studies could be a good example of really integrative multidisciplinary life science in the twenty-first century.     

CdS‐Nanoparticle/Polymer Composite Shells Grown on Silica Nanospheres by Atom‐Transfer Radical Polymerization

In this paper we describe the combined use of surface‐initiated atom transfer radical polymerization (ATRP) and a gas/solid reaction in the direct preparation of CdS‐nanoparticle/block‐copolymer composite shells on silica nanospheres. The block copolymer, consisting of poly(cadmium dimethacrylate) (PCDMA) and poly(methyl methacrylate) (PMMA), is obtained by repeatedly performing the surface‐initiated ATRP procedures in N,N‐dimethylformamide (DMF) solution at room temperature, using cadmium dimethacrylate (CDMA) and methyl methacrylate (MMA) as the monomers. CdS nanoparticles with an average size of about 3 nm are generated in situ by exposing the silica nanospheres coated with block‐copolymer shells to H₂S gas. These synthetic core–shell nanospheres were characterized using transmission electron microscopy (TEM), dynamic light scattering (DLS), thermogravimetric analysis (TGA), diffuse reflectance UV‐vis spectroscopy, X‐ray photoelectron spectroscopy (XPS), and powder X‐ray diffraction (XRD). These composite nanospheres exhibit strong red photoluminescence in the solid state at room temperature.     

人舌鳞癌组织超薄切片的AFM成像和切割

利用一种基于电镜超薄切片法改进的制样方法，将人舌鳞状细胞癌病理组织以环氧树脂包埋并切片后，将薄片平整地贴附在云母上，用原子力显微镜(AFM)对切片表面进行研究，可以得到高分辨率的细胞超微结构图像，局部的亚细胞水平的形态结构可以与电镜下得到的图像相比拟。在此基础上，利用AFM针尖对肿瘤细胞核内特定区域进行切割和操纵，形成生物分子的堆积，从而为拾取(pjck—up)和进一步用分子生物学手段在亚细胞基因水平研究人舌鳞癌的病理学奠定了基础。     

神经内分泌和原始神经外胚叶肿瘤(PNET)的超微结构诊断

本文收集7例神经内分泌和原始神经外胚叶肿瘤,作光镜、免疫组化和电镜观察,参阅文献,侧重讨论了超微结构诊断问题。