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PubMed1969

Mapping of deletions and substitutions in heteroduplex DNA molecules of bacteriophage lambda by electron microscopy.

Electron microscopy of heteroduplex DNA molecules, composed of one strand of Escherichia coli phage lambda(+) DNA annealed to the complementary DNA strand of a lambda deletion or substitution mutant, permits visualization, as well as precise measurements and mapping, of the unpaired single-stranded regions of nonhomology in the otherwise double-stranded molecules. In the lambdab2 mutant, the central segment (13 percent) of the lambda(+) DNA molecule is shown to be deleted. In the hybrid phages lambda(i434) and lambda(i21) a segment of the right arm of the lambda(+) genome (5.5 or 7.6 to 9 percent) is replaced by the corresponding immunity regions of phage 434 (3.3 percent or phage 21 (4 percent) DNA. The b5 region in the lambdab5 mutant appears to be identical to the i(21) segment. From these data it is possible to estimate the size and posiion of those lambda genes which are replaced by the i(434) and i(21) segments. The method permits preparing complete physical maps of viral genomes with a precision heretofore unattainable.

Centrifugation, Density Gradient↗

PubMed1988

Human chromosome banding specific for electron microscopy.

Electron microscopy (EM) provides much higher resolution than that obtained with light microscopy (LM). Until now, however, no chromosome banding procedure specifically adapted for EM was available. To produce an easy and reproducible banding method that would allow accurate chromosome identification, we investigated the applicability of an immunochemical method. BrdU-substituted chromosomal regions can be accurately visualized by applying a monoclonal antibody against BrdU, followed by a gold-tagged secondary antibody. Since BrdU is incorporated during the last part of the S-phase, regions of darkly stained G- and C-bands are substituted. A characteristic C-banding pattern is revealed, and the G-banding obtained is sharp and allows discrimination between subbands. Its similarity with the classical G-banding observed by LM makes it easy to interpret and facilitates karyotyping.

Chromosome Banding↗

PubMed1986

Assembly of chromatin fibers into metaphase chromosomes analyzed by transmission electron microscopy and scanning electron microscopy.

The higher-order assembly of the approximately 30 nm chromatin fibers into the characteristic morphology of HeLa mitotic chromosomes was investigated by electron microscopy. Transmission electron microscopy (TEM) of serial sections was applied to view the distribution of the DNA-histone-nonhistone fibers through the chromatid arms. Scanning electron microscopy (SEM) provided a complementary technique allowing the surface arrangement of the fibers to be observed. The approach with both procedures was to swell the chromosomes slightly, without extracting proteins, so that the densely-packed chromatin fibers were separated. The degree of expansion of the chromosomes was controlled by adjusting the concentration of divalent cations (Mg2+). With TEM, individual fibers could be resolved by decreasing the Mg2+ concentration to 1.0-1.5 mM. The predominant mode of fiber organization was seen to be radial for both longitudinal and transverse sections. Using SEM, surface protuberances with an average diameter of 69 nm became visible after the Mg2+ concentration was reduced to 1.5 mM. The knobby surface appearance was a variable feature, because the average diameter decreased when the divalent cation concentration was further reduced. The surface projections appear to represent the peripheral tips of radial chromatin loops. These TEM and SEM observations support a "radial loop" model for the organization of the chromatin fibers in metaphase chromosomes.

Calcium↗

PubMed1984

Granular cell tumour of the mammary gland simulating malignancy. A report on two cases with light microscopy, transmission electron microscopy and immunohistochemical investigation.

Primary granular cell tumours of the breast in 35 and 55 year old women were studied by light microscopy, transmission electron microscopy and immunohistochemistry. Light and electron microscopy revealed a neural origin of the tumours and this was further substantiated by immunohistochemical studies, with positive S-100 protein reaction and negative reactions for surface heavy and light chains, CEA, alfa-1-antitrypsin, muramidase and GFA-protein. Granular cell tumour of the mammary gland is a very rare tumour. Clinically it sometimes simulates carcinoma because of its fibrous consistency, fixation to pectoral fascia and skin retraction. The diagnosis of granular cell tumour should be included in the differential diagnosis of carcinoma of the breast. The granular cell tumour is derived from neuro-ectodermal tissue. Whether it represents a neurogenic cell-confined metabolic disturbance with lysosomal activation, or a true neoplasm remains to be elucidated.

Adult↗

PubMed2004

Characterization of a Co-CoO obliquely evaporated magnetic tape by analytical electron microscopy and electron holography.

The microstructure and magnetic domain structure of a Co-CoO obliquely evaporated tape for magnetic recording are studied by analytical electron microscopy and electron holography, respectively. While the existence of Co and CoO crystallites is confirmed by energy-filtered electron diffraction, columnar structure of the Co crystallites surrounded by the densely packed CoO crystallites is visualized by an elemental mapping method with electron energy loss spectroscopy, and the crystal orientation relation among the Co crystallites is clarified by high-resolution electron microscopy. It is found that the neighboring Co crystallites have close crystal orientations. On the other hand, electron holography reveals the magnetic flux distribution in a thin section of the tape. Although there exists the background resulting from the effect of inner potential with thickness variation, the distribution of lines of magnetic flux is found to correspond well to the recorded pattern.

Cobalt↗

PubMed1993

A light microscopy, scanning electron microscopy, and laser scanning microscopy analysis of retrieved blade implants after 7 to 20 years of clinical function. A report of 3 cases.

The aim of the present study was to evaluate by means of light microscopy, scanning electron microscopy, and laser scanning microscopy the thin ground sections of blade implants retrieved after 7 to 20 years of clinical function. Microscopic examination revealed that most of the implant surface had an intimate contact with compact lamellar bone tissue. Higher magnifications revealed the presence of a gap (1 to 5 mu) interposed between bone and implant. Many osteocytes were near the implant surface and, in many instances, osteocyte canaliculi, running from the lacunae towards the implant surface, were seen. Structures similar to bone reversal lines were observed at the edge of the bone side of the interface. The bone-titanium region is probably an area of dynamic biological activity.

Aged↗

PubMed1993

[Detecting respiratory syncytial virus in respiratory epithelial cells in adult patients by electron microscopy and immune electron microscopy].

In order to evaluate the value of electron microscopy (EM) in diagnosing respiratory syncytial virus (RSV) infection in adults, the exfoliated cells from nasopharynx of 97 RSV positive patients were examined by EM and the gold labelling technique for immune electron microscopy. The viral particles with RSV features were found at the surface of the exfoliated cells only in a few samples by EM, and the specific gold labelling were observed in the immune electron microscopy (IEM). The results showed that RSV is one of the causative agents for adult respiratory infection.

Adolescent↗

PubMed2004

Methods for in situ detection and characterization of extracellular polymers in biofilms by electron microscopy.

Electron microscopy of biofilms and the localization of extracellular polymers at high resolution require the adaptation of conventional electron microscopic preparation and imaging techniques. A method developed for in situ fixation and embedding of biofilms, imaging of unstained thick sections with electron spectroscopic imaging and the application of lectin or antibody-based marker systems allowed interpretation of extracellular polymer distribution at micrometer scale. By this way, it is possible to discriminate in situ between extracellular polymers produced by different organisms.

Anabaena↗

PubMed1989

Study of the vaginal mucous membrane following tampon utilisation; aspect on colposcopy, scanning electron microscopy and transmission electron microscopy.

The effect of periodic tampons was studied in 17 young women during the menstrual and inter-menstrual cycle. Biopsies (n = 19) were analysed on scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The effect of medium absorbant tampons was compared with that of super-absorbant tampons. On colposcopy, dryness was noted in 89% cases, peeling in 47% cases. Microulcerations were observed only once (5%). Dryness and peeling was most often encountered with the super-absorbant tampons and during the inter-menstrual phase. TEM revealed cellular destruction affecting all layers of the epithelium: lysis of superficial and intermediate cells, destruction of desmosomes accompanied by the creation of inter-cellular spaces, lipidic vacuoles in the lysed cells. SEM revealed epithelial peeling with cleavage and severe cellular desquamation. Cellular anomalies were common at the level of the cytoplasmic membrane, which presented either defects, a porous aspect, or appeared swollen by air-bubbles.

Adult↗

PubMed1988

Innervation of the C cells of chicken ultimobranchial glands studied by immunohistochemistry, fluorescence microscopy, and electron microscopy.

Innervation of the ultimobranchial glands in the chicken was investigated by immunohistochemistry, fluorescence microscopy and electron microscopy. The nerve fibers distributed in ultimobranchial glands were clearly visualized by immunoperoxidase staining with antiserum to neurofilament triplet proteins (200K-, 150K- and 68K-dalton) extracted from chicken peripheral nerves. The ultimobranchial glands received numerous nerve fibers originating from both the recurrent laryngeal nerves and direct vagal branches. The left and right sides of the ultimobranchial region were asymmetrical. The left ultimobranchial gland had intimate contact with the vagus nerve trunk, especially with the distal vagal ganglion, but was somewhat separated from the recurrent nerve. The right gland touched the recurrent nerve, the medial edge being frequently penetrated by the nerve, but the gland was separated from the vagal trunk. The left gland was innervated mainly by the branches from the distal vagal ganglion, whereas the right gland received mostly the branches from the recurrent nerve. The carotid body was located cranially near to the ultimobranchial gland. Large nerve bundles in the ultimobranchial gland ran toward and entered into the carotid body. By fluorescence microscopy, nerve fibers in ultimobranchial glands were observed associated with blood vessels. Only a few fluorescent nerve fibers were present in close proximity to C cell groups; the C cells of ultimobranchial glands may receive very few adrenergic sympathetic fibers. By electron microscopy, numerous axons ensheathed with Schwann cell cytoplasm were in close contact with the surfaces of C cells. In addition, naked axons regarded as axon terminals or "en passant" synapses came into direct contact with C cells. The morphology of these axon terminals and synaptic endings suggest that ultimobranchial C cells of chickens are supplied mainly with cholinergic efferent type fibers. In the region where large nerve bundles and complex ramifications of nerve fibers were present, Schwann cell perikarya investing the axons were closely juxtaposed with C cells; long cytoplasmic processes of Schwann cells encompassed large portions of the cell surface. All of these features suggest that C-cell activity, i.e., secretion of hormones and catecholamines, may be regulated by nerve stimuli.

Animals↗

PubMed1991

Simultaneous visualization of chromosome bands and hybridization signal using colloidal-gold labeling in electron microscopy.

Electron microscopy (EM) is seldom used with in situ hybridization to localize DNA sequences because banding methods for chromosome identification could not be coupled to EM techniques. We have applied an immunochemical replication-banding method specific for EM to solve this problem. A thymidine synchronization/BrdUrd release protocol allows BrdUrd incorporation only into late replicating bands. A biotinylated DNA probe is hybridized in situ to its complementary sequence. The biotinylated probe and the BrdUrd-substituted DNA are simultaneously localized by different reporter/detection systems using different-sized colloidal gold particles as electron-dense tags. We demonstrate the high precision of this mapping procedure by localizing on long prophase chromosomes (greater than 1000 bands per haploid set) the pXBR-1 sequence to a small subregion of the centromeric subband Xp11.1-Xq11.1. This localization to a part of an individual prophase subband is the most precise localization ever reported on human banded mitotic chromosomes.

Cells, Cultured↗

PubMed1997

Correlation of vascular smooth muscle cell morphology observed by scanning electron microscopy with transmission electron microscopy.

Smooth muscle cell (SMC) morphology was studied in rat large mesenteric artery using scanning electron microscopy (SEM), after removal of connective tissue elements with the OsO4/HCl method. Transmission electron microscopy (TEM) was used to correlate the findings of SEM and to assess the effects of digestion on cell ultrastructure. When viewed with SEM, relaxed SMC possessed a smooth, regular surface morphology with longitudinal striations or grooves. TEM examination of cross-sectional profiles of SMC suggested that these striations corresponded to the arrangement of membrane dense bodies within the SMC. The presence of cell contacts among SMC as observed with SEM were confirmed using TEM. Two types of SMC contacts were found: simple appositional contacts and interdigitations. Gap junctions were not observed between SMC in the media. Although structures suggesting nerve networks were observed in the medial-adventitial border with SEM, this could not be confirmed using TEM as a result of alteration in cell ultrastructure with the digestive method. We conclude that the OsO4/HCl method, although useful for SEM, is not suitable for TEM study, because it caused significant alterations of SMC ultrastructure such that organelles and other structures were no longer evident when viewed with TEM.

Animals↗

PubMed2006

Scanning electron microscopy and transmission electron microscopy study of hot-deformed gamma-TiAl-based alloy microstructure.

The aim of this work was to assess the changes in the microstructure of hot-deformed specimens made of alloys containing 46-50 at.% Al, 2 at.% Cr and 2 at.% Nb (and alloying additions such as carbon and boron) with the aid of scanning electron microscopy and transmission electron microscopy techniques. After homogenization and heat treatment performed in order to make diverse lamellae thickness, the specimens were compressed at 1000 degrees C. Transmission electron microscopy examinations of specimens after the compression test revealed the presence of heavily deformed areas with a high density of dislocation. Deformation twins were also observed. Dynamically recrystallized grains were revealed. For alloys no. 2 and no. 3, the recovery and recrystallization processes were more extensive than for alloy no. 1.

Journal Article↗

PubMed1998

Investigation of cholesterol, bilirubin, and protein distribution in human gallstones by color cathodoluminescence scanning electron microscopy and transmission electron microscopy.

The application of color cathodoluminescent scanning electron microscopy (CCL-SEM) for qualitative luminescence analysis of cholesterol, bilirubin, and protein in human gallastones was demonstrated. Images of these deposits (cholesterol, bilirubin, and protein) were formed in real colors (blue-cholesterol, red, orange-bilirubin, yellow, green-protein) in accordance with the cathodoluminescent spectrum for each control material. The other method described for transmission electron microscopy (TEM) of ultrathin sections provides more detailed characterization of the ultrastructure of cholesterol-containing regions and their spatial interrelations with bilirubin-containing regions. Using CCL-SEM combined with TEM permits the receipt of more complete information about the chemical composition and ultrastructure of gallstones and may lead to more effective understanding of the pathogenesis of cholesterol cholelithiasis.

Bilirubin↗

PubMed1987

Tumor vasculature in young and old hosts: scanning electron microscopy of microcorrosion casts with microangiography, light microscopy and transmission electron microscopy.

Tumor growth in vivo is dependent upon new blood vessel formation. When B16-F10 melanoma cells are implanted subcutaneously in young (3 mo) and old (24 mo) C57BL/6 mice the rate of growth is dependent on the age of the mice. This study involved a wide range of histological and microscopic techniques but was limited primarily to the initial phase of tumor growth. Stereological point counting from light microscopy (LM) of standard histological sections has been used to yield data regarding blood content. Tumor-bearing mice were perfused through the aorta with a fixation solution and were infused with a low-viscosity radiopaque gel (Microfil) or resin (Mercox). Soft x-rays of the whole animal were used for identifying the feeding vessels to the tumor. Tumors with Microfil were sliced and used for microangiography and light-microscopic observation while those with resin were used to make corrosion casts for scanning electron microscopy (SEM). The different characteristics of the tumor blood vessels in different aged mice were most obvious through SEM of vascular corrosion casts. In comparison with tumors in young mice those of similar size in old hosts had more necrosis, reduced presence of angiogenic features, decreased vessel density, reduced penetration into the tumor, and enhanced tortuosity of the vessel lumen. Transmission electron microscopy (TEM) revealed incompletely developed wall structure of the vessels regardless of host. The above results are consistent with the hypothesis that retarded angiogenesis may be responsible in part for the limited growth of tumors in old hosts.

Animals↗

PubMed2006

Uptake of C60 by human monocyte macrophages, its localization and implications for toxicity: studied by high resolution electron microscopy and electron tomography.

Despite great interest in the engineering applications of carbon-based nanoparticles, recent studies have raised concerns about their potential toxicity and safety. The release of C(60) into the environment has been suggested to be a potential risk with possible ecological implications. Here we evaluate energy-filtered transmission electron microscopy (EFTEM) and scanning transmission electron microscopy (STEM)-based electron tomography as techniques for imaging the three-dimensional (3-D) distribution of nanoparticles within cells. Our aim was to establish if human monocyte macrophages internalise nanoparticles and to assess whether nanoparticles are modified by cells following uptake. Using these techniques we were able to show a marked increased in the amount of information gained from 3-D imaging. 3-D electron tomography revealed several sub-cellular compartments containing C(60) within the cell: secondary lysosomes, along the outer and nuclear membrane and most notably inside the nucleus of the cell. Using EFTEM and STEM-based techniques we were able to visualize cell structures such as membranes, the mitochondria, ribosomes and the nucleus, without the need for traditional staining techniques. In particular we demonstrate the potential of electron tomography for whole cell studies to enable 3-D distributions of particles within cells. The concentrations of C(60) used in this study were not toxic and were chosen to study which sub-cellular compartments accumulated C(60). Knowledge of the sites of accumulation of nanoparticles will allow us to predict vulnerability if the nanoparticles can generate free radicals.

Cell Culture Techniques↗

PubMed1985

Scanning electron microscopy and transmission electron microscopy of the ciliated cells of the trachea of the rabbit treated with misonidazole alone and in combination with ionizing radiation.

The trachea is often located in the treatment volume when irradiating malignant tumours in the thorax. In order to evaluate possible synergism between misonidazole and irradiation on this tissue, the following studies were made. Fifty rabbits were treated with daily injections of 100 mg misonidazole given i.p. on consecutive days from 1 to 10 days. Morphological investigations of the trachea were made with scanning electron microscopy (SEM), transmission electron microscopy (TEM) and light microscopy (LM). Physiological examinations were performed with recording of the ciliary beat frequency. The results were compared with those from a group of 100 rabbits given misonidazole in a similar manner and exposed to irradiation (2 Gy) 15-30 min after each injection. Ten rabbits were used as controls. The results are compared to the effect of fractionated irradiation alone with 2 Gy/day. Fractionated irradiation of the ciliary epithelium in the trachea of the rabbit has shown dose-dependent physiological and morphological effects. Misonidazole potentiates these effects of radiation with a more pronounced change of the ciliary beat frequency and an increased metabolic activity as could be visualized on TEM. The combination of drug and irradiation also induced a hyperplasia of the ciliary epithelium. Misonidazole itself had no effect on the ciliary beat frequency, but caused a hypoplasia of the ciliary epithelium.

Animals↗

PubMed2006

Electronic structure analyses of BN network materials using high energy-resolution spectroscopy methods based on transmission electron microscopy.

Electronic structures of boron-nitride (BN) nanotubes and a BN cone-structure material were studied by using a high energy-resolution electron energy-loss spectroscopy (EELS) microscope. A trial of the whole electronic structure study of hexagonal BN (h-BN), which consists of flat BN honeycomb layers, was conducted by a combination of EELS and X-ray emission spectroscopy (XES) based on transmission electron microscopy (TEM) (TEM-EELS/XES). The pi and pi+sigma plasmon energies of BN nanotubes (BNT) were smaller than those of h-BN. The pi+sigma energy was explained by the surface plasmon excitation. The spectrum of a two-wall BNT of 2.7 nm in diameter showed a new spectral onset at 4 eV. The valence electron excitation spectra obtained from the tip region of the BN cone with an apex angle of 20 degrees showed similar intensity distribution with those of BNTs. The B K-shell electron excitation spectra obtained from the bottom edge region of the BN cone showed additional peak intensity when compared with those of h-BN and BNT. The B K-shell electron excitation spectra and B K-emission spectra of h-BN were compared with a result of a LDA band calculation. It showed that high symmetry points in the band diagram appear as peak and/or shoulder structures in the EELS and XES spectra. Interband transitions appeared in the imaginary part of the dielectric function of h-BN experimentally obtained were assigned in the band diagram. The analysis also presented that the LDA calculation estimated the bandgap energy smaller than the real material by an amount of 2 eV. Those results of TEM-EELS/XES analysis presented that high energy-resolution spectroscopy methods combined with TEM is a promising method to analyze whole electronic structures of nanometer scale materials.

Boron↗