Electron Microscope Study of RNA&#39;s of Paramyxoviruses

Sendai Virus ◽

Sodium Dodecyl ◽

Rna Molecules ◽

Virus Particles ◽

Molecular Weights ◽

Length Measurements

The ribonucleic acid (RNA) of paramyxoviruses has been characterized by biochemical and physiochemical methods. However, paramyxovirus RNA molecules have not been studied by electron microscopy. The molecular weights of these single-stranded viral RNA molecules are not known as yet. Since electron microscopy has been found to be useful for the characterization of single-stranded RNA, this investigation was initiated to examine the morphology and length measurements of paramyxovirus RNA's.Sendai virus Z strain and Newcastle disease virus (NDV), Milano strain, were used. For these studies it was necessary to develop a method of extracting RNA molecules from purified virus particles. Highly purified Sendai virus was treated with pronase (300 μg/ml) at 37°C for 30 minutes and the RNA extracted by the sodium dodecyl sulfate (SDS)-phenol procedure.

An Electron Microscope Study of Interdiffusion and Compound Formation in Ta-Au Thin Films

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100070606 ◽

1973 ◽

Vol 31 ◽

pp. 32-33 ◽

Cited By ~ 1

Author(s):

T. C. Tisone ◽

S. Lau

Keyword(s):

Electron Microscope ◽

Thermally Grown Oxide ◽

Ion Milling ◽

Compound Formation ◽

Technology Application ◽

Transmission Electron ◽

Before And After ◽

Au Thin Films

In a study of the properties of a Ta-Au metallization system for thin film technology application, the interdiffusion between Ta(bcc)-Au, βTa-Au and Ta2M-Au films was studied. Considered here is a discussion of the use of the transmission electron microscope(TEM) in the identification of phases formed and characterization of the film microstructures before and after annealing.The films were deposited by sputtering onto silicon wafers with 5000 Å of thermally grown oxide. The film thicknesses were 2000 Å of Ta and 2000 Å of Au. Samples for TEM observation were prepared by ultrasonically cutting 3mm disks from the wafers. The disks were first chemically etched from the silicon side using a HNO3 :HF(19:5) solution followed by ion milling to perforation of the Au side.

Nucleic Acid Molecules Prepared by Monolayer Techniques

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100094140 ◽

1972 ◽

Vol 30 ◽

pp. 178-179

Author(s):

Dimitrij Lang

Keyword(s):

Electron Microscopy ◽

Standard Deviation ◽

Nucleic Acid ◽

Cytochrome C ◽

Nucleic Acids ◽

Contour Length ◽

Sample Standard Deviation ◽

Molecular Weights ◽

Length Measurements ◽

Protein Monolayer

The success of the protein monolayer technique for electron microscopy of individual DNA molecules is based on the prevention of aggregation and orientation of the molecules during drying on specimen grids. DNA adsorbs first to a surface-denatured, insoluble cytochrome c monolayer which is then transferred to grids, without major distortion, by touching. Fig. 1 shows three basic procedures which, modified or not, permit the study of various important properties of nucleic acids, either in concert with other methods or exclusively:1) Molecular weights relative to DNA standards as well as number distributions of molecular weights can be obtained from contour length measurements with a sample standard deviation between 1 and 4%.

Characterization of submicrometer fluid Inclusions in minerals by Analytical/Transmission Electron Microscopy and electron diffraction

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100175259 ◽

1990 ◽

Vol 48 (4) ◽

pp. 424-424

Author(s):

George Guthrie ◽

David Veblen

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Electron Microscope ◽

Electron Diffraction ◽

Light Microscopy ◽

Fluid Inclusions ◽

Energy Dispersive Spectrometer ◽

Analytical Transmission Electron Microscopy ◽

Transmission Electron

The nature of a geologic fluid can often be inferred from fluid-filled cavities (generally <100 μm in size) that are trapped during the growth of a mineral. A variety of techniques enables the fluids and daughter crystals (any solid precipitated from the trapped fluid) to be identified from cavities greater than a few micrometers. Many minerals, however, contain fluid inclusions smaller than a micrometer. Though inclusions this small are difficult or impossible to study by conventional techniques, they are ideally suited for study by analytical/ transmission electron microscopy (A/TEM) and electron diffraction. We have used this technique to study fluid inclusions and daughter crystals in diamond and feldspar.Inclusion-rich samples of diamond and feldspar were ion-thinned to electron transparency and examined with a Philips 420T electron microscope (120 keV) equipped with an EDAX beryllium-windowed energy dispersive spectrometer. Thin edges of the sample were perforated in areas that appeared in light microscopy to be populated densely with inclusions. In a few cases, the perforations were bound polygonal sides to which crystals (structurally and compositionally different from the host mineral) were attached (Figure 1).

An Electron Microscope Study of Basophile Substances of Frozen-Dried Rat Liver

10.1083/jcb.4.3.291 ◽

1958 ◽

Vol 4 (3) ◽

pp. 291-300 ◽

Cited By ~ 25

Author(s):

Henry Finck

Keyword(s):

Electron Microscopy ◽

Electron Microscope ◽

Rat Liver ◽

Microscope Study ◽

Freeze Drying ◽

Light And Electron Microscopy ◽

Enzyme Treatment ◽

Dense Granules ◽

Homogeneous Matrix

Small pieces of liver from rats subjected to different dietary regimes were fixed by freeze-drying, and postfixed by in vacuo heating and denaturation with alcohol. Specimens were digested with ribo- or deoxyribonuclease, and stained with gallocyanin-chromalum, azure II, the Feulgen procedure or alcoholic platinic tetrabromide. Some specimens were reserved as controls of the effects of enzyme treatment. Stained and unstained specimens were embedded in methacrylate and examined by light and electron microscopy. Basophilic and Feulgen-positive substances, after contact with watery reagents, were found by electron microscopy to exist as small dense granules embedded in a less dense homogeneous matrix, forming the walls of submicroscopic vacuoles. These granules were absent after digestion with nucleodepolymerases. In specimens (unstained, or stained with platinic tetrabromide) which had not passed through water, the dense (basophile) substances in nuclei and cytoplasm were found to exist, not as granules, but as ill defined submicroscopic concentrates which blended imperceptibly into the homogeneous matrix of the vacuolar walls. Objections to the use of stains for improving contrast conditions in electron microscopy of tissues are discussed, and it is concluded that the reagents do not necessarily produce the observed increases in contrast by selectively stabilizing certain structures. The concept of microsomes as pre-existing distinct morphological entities in intact (unhomogenized) cells is thought to be inconsistent with the distribution of basophile substances in frozen-dried liver.

ELECTRON MICROSCOPY OF THE NUCLEAR MEMBRANE OF AMOEBA PROTEUS

10.1083/jcb.2.4.445 ◽

1956 ◽

Vol 2 (4) ◽

pp. 445-448 ◽

Cited By ~ 60

Author(s):

Marie H. Greider ◽

Wencel J. Kostir ◽

Walter J. Frajola

Keyword(s):

Electron Microscopy ◽

Electron Microscope ◽

Nuclear Membrane ◽

Outer Layer ◽

Microscope Study ◽

Cell Types ◽

Amoeba Proteus ◽

Nuclear Membranes ◽

Thin Sectioning

An electron microscope study of the nuclear membrane of Amoeba proteus by thin sectioning techniques has revealed an ultrastructure in the outer layer of the membrane that is homologous to the pores and annuli observed in the nuclear membranes of many other cell types studied by these techniques. An inner honeycombed layer apparently unique to Amoeba proteus is also described.

Effect of detergents and chemicals on purified vaccinia virus: analysis by SDS polyacrylamide gel electrophoresis and electron microscopy

Canadian Journal of Microbiology ◽

10.1139/m77-036 ◽

1977 ◽

Vol 23 (3) ◽

pp. 240-252 ◽

Cited By ~ 4

Author(s):

J. Boisvert ◽

T. Yamamoto

Keyword(s):

Electron Microscopy ◽

Vaccinia Virus ◽

Gel Electrophoresis ◽

Alkali Treatment ◽

Guanidine Hydrochloride ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Ammonium Bromide ◽

Molecular Weights ◽

Viral Particles

Vaccinia virus particles were dissociated into their constituent polypeptides and analysed by sodium dodecyl sulfate (SDS) gel electrophoresis. Thirty-three distinct polypeptide bands were identified and their molecular weights ranged between 11 000 and 150 000 daltons.Specific staining of gels containing polypeptides of dissociated virions revealed the presence of eight glycopeptides. No lipopeptides were detected.Analysis of chemical extracts (urea, guanidine hydrochloride, and alkali treatment) of the virus by SDS gel electrophoresis indicated that a total of 10 to 14 different polypeptides ranging in molecular weights from 11 000 to 70 000 daltons were solubilized.Analysis of detergent extracts and of the remains of extracted viral particles has shown that the detergent Nonidet P-40 (NP-40) solubilized a total of 11 polypeptides of which 6 were glycopeptides. The other detergents sodium deoxycholate (SDC) and cetyl trimethyl ammonium bromide (CTAB) were not as selective, both solubilizing more than 25 of the polypeptides composing the virus. Gel electrophoresis results also indicated that most of the small molecular weight (11 000–70 000 daltons) polypeptides were readily solubilized by NP-40, SDC, and CTAB, while those with molecular weights of 70 000 daltons and higher were not well solubilized.The effects of detergents were also analysed by electron microscopy. Evidence was obtained for subpopulations of viral particles having different susceptibility to detergent extraction.

AN ELECTRON MICROSCOPE STUDY OF THE EMBRYOLOGY OF THE INTERCALATED DISC IN THE HEART OF THE RABBIT

10.1083/jcb.3.2.193 ◽

1957 ◽

Vol 3 (2) ◽

pp. 193-202 ◽

Cited By ~ 84

Author(s):

Alan R. Muir

Keyword(s):

Electron Microscopy ◽

Electron Microscope ◽

Cardiac Muscle ◽

Microscope Study ◽

Light Microscope ◽

Muscle Cells ◽

Intercalated Disc ◽

Adult Cell ◽

Embryonic Muscle

Prenatal and postnatal cardiac muscle from rabbits has been studied by electron microscopy, after osmium fixation and methacrylate embedding. The observations showed that 1. Cell membranes divide the muscle into cellular units from the youngest embryo which was studied (9½ days after coitus) until the adult state. 2. The embryonic muscle cells contain only one nucleus, whereas the adult cell may be multinucleated. 3. At all stages of development, wherever a myofibrillar axis crosses a cellular boundary, the myofilaments are interrupted by an intercalated disc. 4. With age, increase in size and complexity of the discs render them recognisable by the light microscope.

Purification and Characterization of 2,6-β-d-Fructan 6-Levanbiohydrolase fromStreptomyces exfoliatus F3-2

Applied and Environmental Microbiology ◽

10.1128/aem.66.1.252-256.2000 ◽

2000 ◽

Vol 66 (1) ◽

pp. 252-256 ◽

Cited By ~ 19

Author(s):

Katsuichi Saito ◽

Kazuya Kondo ◽

Ichiro Kojima ◽

Atsushi Yokota ◽

Fusao Tomita

Keyword(s):

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Gel Filtration ◽

Extracellular Enzyme ◽

Molecular Weights ◽

Sds Page ◽

Monomeric Structure ◽

Ph Range ◽

Hydrolysis Of

ABSTRACT Streptomyces exfoliatus F3-2 produced an extracellular enzyme that converted levan, a β-2,6-linked fructan, into levanbiose. The enzyme was purified 50-fold from culture supernatant to give a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weights of this enzyme were 54,000 by SDS-PAGE and 60,000 by gel filtration, suggesting the monomeric structure of the enzyme. The isoelectric point of the enzyme was determined to be 4.7. The optimal pH and temperature of the enzyme for levan degradation were pH 5.5 and 60°C, respectively. The enzyme was stable in the pH range 3.5 to 8.0 and also up to 50°C. The enzyme gave levanbiose as a major degradation product from levan in an exo-acting manner. It was also found that this enzyme catalyzed hydrolysis of such fructooligosaccharides as 1-kestose, nystose, and 1-fructosylnystose by liberating fructose. Thus, this enzyme appeared to hydrolyze not only β-2,6-linkage of levan, but also β-2,1-linkage of fructooligosaccharides. From these data, the enzyme from S. exfoliatus F3-2 was identified as a novel 2,6-β-d-fructan 6-levanbiohydrolase (EC 3.2.1.64 ).

Immunofluorescence and electron microscopy of the cytoplasmic surface of the human erythrocyte membrane and its interaction with Sendai virus.

10.1083/jcb.83.2.338 ◽

1979 ◽

Vol 83 (2) ◽

pp. 338-347 ◽

Cited By ~ 29

Author(s):

M Büechi ◽

T Bächi

Keyword(s):

Electron Microscopy ◽

Plasma Membrane ◽

Plasma Membranes ◽

Sendai Virus ◽

Light And Electron Microscopy ◽

Distal Portion ◽

Double Labeling ◽

Virus Particles ◽

Viral Antigens ◽

Cytoplasmic Surface

A method was developed for directly observing the inner surfaces of plasma membranes by light and electron microscopy. Human erythrocytes were attached to cover slips (glass or mica) treated with aminopropylsilane and glutaraldehyde, and then disrupted by direct application of a jet of buffer, which removed the distal portion of the cells, thus exposing the cytoplasmic surface (PS) of the flattened membranes. Antispectrin antibodies and Sendai virus particles were employed as sensitive markers for, respectively, the PS and the external surface (ES) of the membrane; their localization by immunofluorescence or electron microscopy demonstrated that the major asymmetrical features of the plasma membrane were preserved. The fusion of Sendai virus particles with cells was investigated using double-labeling immunofluorescence techniques. Virus adsorbed to the ES of cells at 4 degrees C was not accessible to fluorescein-labeled antibodies applied from the PS side. After incubation at 37 degrees C, viral antigens could be detected at the PS. These antigens, however, remained localized and did not diffuse from the site of attachment, as is usually seen in viral antigens accessible on the ES. They may therefore represent internal viral antigens not incorporated into the plasma membrane as a result of virus-cell fusion.

ELECTRON MICROSCOPY OF PLASMA-CELL TUMORS OF THE MOUSE

10.1083/jcb.9.2.353 ◽

1961 ◽

Vol 9 (2) ◽

pp. 353-368 ◽

Cited By ~ 31

Author(s):

D. F. Parsons ◽

E. B. Darden ◽

D. L. Lindsley ◽

Guthrie T. Pratt

Keyword(s):

Electron Microscopy ◽

Endoplasmic Reticulum ◽

Plasma Cell ◽

Plasma Cells ◽

Granular Body ◽

Virus Particles ◽

Cytoplasmic Structure ◽

Large Numbers ◽

C3h Mice

An electron microscope study was made of a series of transplanted MPC-1 plasma-cell tumors carried by BALB/c mice. Large numbers of particles similar in morphology to virus particles were present inside the endoplasmic reticulum of tumor plasma cells. Very few particles were seen outside the cells or in ultracentrifuged preparations of the plasma or ascites fluid. In very early tumors particles were occasionally seen free in the cytoplasm adjacent to finely granular material. In general, the distribution of these particles inside endoplasmic reticulum is similar in early and late tumors. A few transplanted X5563 tumors of C3H mice were also examined. Large numbers of particles were found in the region of the Golgi apparatus in late X5663 tumors. A newly described cytoplasmic structure of plasma cells, here called a "granular body," appears to be associated with the formation of the particles. Particles present in MPC-1 tumors are exclusively of a doughnut form, whereas some of those in the inclusions of the late X5563 tumors show a dense center. Normal plasma cells, produced by inoculation of a modified Freund adjuvant into BALB/c mice. have been compared morphologically with tumor plasma cells of both tumor lines.