Histology and ultrastructure of oat blue dwarf virus infected oats

1972 ◽  
Vol 50 (12) ◽  
pp. 2511-2520 ◽  
Author(s):  
R. J. Zeyen ◽  
E. E. Banttari
Keyword(s):  
Light Microscopy ◽  
Sieve Tube ◽  
Dwarf Virus ◽  
Electron Microscopic ◽  
Virus Particles ◽  
Procambial Strand ◽  
Sieve Plates ◽  
Aster Leafhopper ◽  
Sieve Tube Element

The oat blue dwarf virus is a small spherical virus (28–30 nm) in diameter and is obligatorily transmitted by the aster leafhopper Macrosteles fascifrons Stål. The virus causes abnormalities in the phloem development of infected plants. Hyperplasia and limited hypertrophy of phloic procambium, in a given procambial strand, begin only after the maturation of the first protophloem sieve-tube element in that particular localized area. The majority of phloem elements produced in hyperplastic areas are parenchymatous, have truncated end walls, and lack sieve plates. Electron-microscopic observations substantiated the hypothesis that the virus was phloem-limited by revealing virus particles only in phloem elements. The greatest accumulation of virus particles was observed in the region between immature and fully vacuolated phloem elements, implicating virus synthesis in immature elements. Crystals of virus particles were often large enough to be seen by light microscopy.

scholarly journals RELATION OF BEET YELLOWS VIRUS TO THE PHLOEM AND TO MOVEMENT IN THE SIEVE TUBE

1967 ◽  
Vol 32 (1) ◽  
pp. 71-87 ◽  
Author(s):  
K. Esau ◽  
J. Cronshaw ◽  
L. L. Hoefert
Keyword(s):  
Sieve Tube ◽  
Plant Viruses ◽  
Passive Transport ◽  
Sieve Elements ◽  
Virus Particles ◽  
Parenchyma Cells ◽  
Beet Yellows Virus ◽  
Sieve Tubes ◽  
Sieve Plates ◽  
Orderly Arrangement

In minor veins of leaves of Beta vulgaris L. (sugar beet) yellows virus particles were found both in parenchyma cells and in mature sieve elements. In parenchyma cells the particles were usually confined to the cytoplasm, that is, they were absent from the vacuoles. In the sieve elements, which at maturity have no vacuoles, the particles were scattered throughout the cell. In dense aggregations the particles tended to assume an orderly arrangement in both parenchyma cells and sieve elements. Most of the sieve elements containing virus particles had mitochondria, plastids, endoplasmic reticulum, and plasma membrane normal for mature sieve elements. Some sieve elements, however, showed evidence of degeneration. Virus particles were present also in the pores of the sieve plates, the plasmodesmata connecting the sieve elements with parenchyma cells, and the plasmodesmata between parenchyma cells. The distribution of the virus particles in the phloem of Beta is compatible with the concept that plant viruses move through the phloem in the sieve tubes and that this movement is a passive transport by mass flow. The observations also indicate that the beet yellows virus moves from cell to cell and in the sieve tube in the form of complete particles, and that this movement may occur through sieve-plate pores in the sieve tube and through plasmodesmata elsewhere.

Sieve tube elements in the stem of Neptunia oleracea Lour

1968 ◽  
Vol 16 (3) ◽  
pp. 433 ◽  
Author(s):  
JJ Shah ◽  
MR James
Keyword(s):  
Aquatic Plant ◽  
Sieve Tube ◽  
Sieve Plate ◽  
Callose Deposition ◽  
Nuclear Disintegration ◽  
Full Development ◽  
Companion Cells ◽  
Sieve Plates ◽  
Structural Aspects ◽  
Sieve Tube Element

Some structural aspects of the phloem of Neptunia oleracea, an aquatic plant, are reported. The sieve tube elements on an average are 190μ long and 13μ wide and have compound sieve plates at varying degrees of inclination. The developing sieve tube element has a single large spindle-shaped slime body, which presumably has an outer membrane. The slime body undergoes dispersal before or after full development of the sieve plate, but often nuclear degeneration occurs first. Distinct slime plugs are absent. Plastids and other granular bodies are attached to many of the strands, which are less than 0.5μ in diameter. During the process of nuclear disintegration the nuclear membrane is indistinct, and extruded nucleolus is not observed. Sieve areas and connections are comparatively few in number, and the sieve areas and wall connections as well as the sieve plates show scanty callose deposition. Plastids are abundant in the sieve tube elements, especially near the sieve plates. The companion cells of two consecutive sieve tube elements are placed on alternate sides and hence their longitudinal continuity is not always maintained. Companion cells do not exceed the length of the sieve tube element.

Further Observations on the Virus of Epizootic Diarrhea of Infant Mice. An Electron Microscopic Study

1967 ◽  
Vol 25 ◽  
pp. 110-111
Author(s):  
W. G. Banfield ◽  
G. Kasnic ◽  
J. H. Blackwell
Keyword(s):  
Electron Microscope ◽  
Infected Cell ◽  
Microscopic Study ◽  
Intestinal Epithelium ◽  
Ultrastructural Study ◽  
Osmium Tetroxide ◽  
Lead Citrate ◽  
Electron Microscopic ◽  
Virus Particles ◽  
Infected Cells

An ultrastructural study of the intestinal epithelium of mice infected with the agent of epizootic diarrhea of infant mice (EDIM virus) was first performed by Adams and Kraft. We have extended their observations and have found developmental forms of the virus and associated structures not reported by them.Three-day-old NLM strain mice were infected with EDIM virus and killed 48 to 168 hours later. Specimens of bowel were fixed in glutaraldehyde, post fixed in osmium tetroxide and embedded in epon. Sections were stained with uranyl magnesium acetate followed by lead citrate and examined in an updated RCA EMU-3F electron microscope.The cells containing virus particles (infected) are at the tips of the villi and occur throughout the intestine from duodenum through colon. All developmental forms of the virus are present from 48 to 168 hours after infection. Figure 1 is of cells without virus particles and figure 2 is of an infected cell. The nucleus and cytoplasm of the infected cells appear clearer than the cells without virus particles.

Immunocytochemistry. A Review of Methodology for Electron Microscopic Applicaiton

1974 ◽  
Vol 32 ◽  
pp. 328-329
Author(s):  
Joseph E. Mazurkiewicz
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Ultrastructural Level ◽  
Electron Microscopic ◽  
Biological Interest ◽  
Investigative Approach ◽  
Immunologic Activity ◽  
Dense Label

Immunocytochemistry is a powerful investigative approach in which one of the most exacting examples of specificity, that of the reaction of an antibody with its antigen, isused to localize tissue and cell specific molecules in situ. Following the introduction of fluorescent labeled antibodies in T950, a large number of molecules of biological interest had been studied with light microscopy, especially antigens involved in the pathogenesis of some diseases. However, with advances in electron microscopy, newer methods were needed which could reveal these reactions at the ultrastructural level. An electron dense label that could be coupled to an antibody without the loss of immunologic activity was desired.

A Method for Electron Microscopic Study of Tissue Culture Cell Monolayers Grown in Tubes

1967 ◽  
Vol 25 ◽  
pp. 132-133
Author(s):  
R. Stephens ◽  
G. Schidlovsky ◽  
S. Kuzmic ◽  
P. Gaudreau
Keyword(s):  
Electron Microscopy ◽  
Tissue Culture ◽  
Light Microscopy ◽  
Cell Sheet ◽  
Culture Cell ◽  
Tissue Culture Cell ◽  
Electron Microscopic ◽  
Cell Sheets ◽  
Morphological Alterations ◽  
Culture Cells

The usual method of scraping or trypsinization to detach tissue culture cell sheets from their glass substrate for further pelletization and processing for electron microscopy introduces objectionable morphological alterations. It is also impossible under these conditions to study a particular area or individual cell which have been preselected by light microscopy in the living state.Several schemes which obviate centrifugation and allow the embedding of nondetached tissue culture cells have been proposed. However, they all preserve only a small part of the cell sheet and make use of inverted gelatin capsules which are in this case difficult to handle.We have evolved and used over a period of several years a technique which allows the embedding of a complete cell sheet growing at the inner surface of a tissue culture roller tube. Observation of the same cell by light microscopy in the living and embedded states followed by electron microscopy is performed conveniently.

Correlative video enhanced differential interference contrast light microscopy (VDIC)-LM), low-voltage high-resolution SEM (LV-HR-SEM), and high-voltage electron microscopy (HVEM) in the observation of gold-labelled surface antigens and the subjacent cytoskeletal matrix

1989 ◽  
Vol 47 ◽  
pp. 904-905
Author(s):  
Ralph M. Albrecht ◽  
Scott R. Simmons ◽  
Marek Malecki
Keyword(s):  
High Resolution ◽  
Light Microscopy ◽  
Low Voltage ◽  
Microscopic Analysis ◽  
Cell Labelling ◽  
Video Technology ◽  
Electron Microscopic ◽  
Image Capture ◽  
High Voltage Electron Microscopy ◽  
Fluorescence Studies

The development of video-enhanced light microscopy (LM) as well as associated image processing and analysis have significantly broadened the scope of investigations which can be undertaken using (LM). Interference/polarization based microscopies can provide high resolution and higher levels of “detectability” especially in unstained living systems. Confocal light microscopy also holds the promise of further improvements in resolution, fluorescence studies, and 3 dimensional reconstruction. Video technology now provides, among other things, a means to detect differences in contrast difficult to detect with the human eye; furthermore, computerized image capture, processing, and analysis can be used to enhance features of interest, average images, subtract background, and provide a quantitative basis to studies of cells, cell features, cell labelling, and so forth. Improvements in video technology, image capture, and cost-effective computer image analysis/processing have contributed to the utility and potential of the various interference and confocal microscopic instrumentation.Electron microscopic technology has made advances as well. Microprocessor control and improved design have contributed to high resolution SEMs which have imaging capability at the molecular level and can operate at a range of accelerating voltages starting at 1KV. Improvements have also been seen in the HVEM and IVEM transmission instruments. As a whole, these advances in LM and EM microscopic technology provide the biologist with an array of information on structure, composition, and function which can be obtained from a single specimen. Corrrelative light microscopic analysis permits examination of living specimens and is critical where the “history” of a cell, cellular components, or labels needs to be known up to the time of chemical or physical fixation. Features such as cytoskeletal elements or gold label as small as 0.01 μm, well below the 0.2 μm limits of LM resolution, can be “detected” and their movement followed by VDIC-LM. Appropriate identification and preparation can then lead to the examination of surface detail and surface label with stereo LV-HR-SEM. Increasing the KV in the HR-SEM while viewing uncoated or thinly coated specimens can provide information from beneath the surface as well as increasing Z contrast so that positive identification of surface and subsurface colloidal gold or other heavy metal labelled/stained material is possible. Further examination of the same cells using stereo HVEM or IVEM provides information on internal ultrastructure and on the relationship of labelled material to cytoskeletal or organellar distribution, A wide variety of investigations can benefit from this correlative approach and a number of instrumentational configurations and preparative pathways can be tailored for the particular study. For a surprisingly small investment in time and technique, it is often possible to clear ambiguities or questions that arise when a finding is presented in the context of only one modality.

Electron Microscopic Study of the Graffi Virus in Bone Marrow of Leukemic Mice.

1966 ◽  
Vol 52 (1) ◽  
pp. 35-59 ◽  
Author(s):  
Natale Pennelli ◽  
Luciano Fiore-Donati ◽  
Luigi Chieco-Bianchi ◽  
Giuseppe Tridente
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Electron Microscopic ◽  
Virus Particles ◽  
Negative Stain ◽  
Cytoplasmic Vesicles ◽  
C57bl Mice ◽  
Ultrathin Sections ◽  
Number Of Particles ◽  
Marrow Cells

Bone marrow from C57BL mice with myeloid leukemia induced by Graffi virus has been studied with the electron microscope by ultrathin section and negative stain techniques. Virus particles were usually found in different types of bone marrow cells as well as in extracellular spaces. However, the highest number of particles in various stages of maturation was observed in the cytoplasm of megakaryocytes. Two main types of virus particle were found: the immature Al particle and the mature C particle. They were morphologically indistinguishable from other murine leukemogenic viruses. In partially purified preparations studied by negative staining, some of the particles which were not penetrated by PTA, frequently showed a tail-like structure of variable length. In ultrathin sections, particles were found to originate by budding from the cell membranes. Budding of particles was particularly evident in megakaryocytes and especially within the granules and cytoplasmic vesicles or in connection with the platelet demarcating membranes. The findings of a high number of virus particles in all stages of maturation in megakaryocytes together with a certain degree of megakaryocytosis observed in the bone marrow suggest that this type of cell is possibly one of the main source of production of the virus. A few particles resembling morphologically mycoplasma were detected within the cytoplasm of some immature bone marrow cells.

Hepatitis E virus egress depends on the exosomal pathway, with secretory exosomes derived from multivesicular bodies

2014 ◽  
Vol 95 (10) ◽  
pp. 2166-2175 ◽  
Author(s):  
Shigeo Nagashima ◽  
Suljid Jirintai ◽  
Masaharu Takahashi ◽  
Tominari Kobayashi ◽  
Tanggis ◽  
...  
Keyword(s):  
Hepatitis E Virus ◽  
Lipid Membrane ◽  
Hepatitis E ◽  
Negative Control ◽  
Bafilomycin A1 ◽  
Genotype 3 ◽  
Electron Microscopic ◽  
Immunogold Staining ◽  
Virus Particles ◽  
Virion Release

Our previous studies indicated that hepatitis E virus (HEV) forms membrane-associated particles in the cytoplasm, most likely by budding into intracellular vesicles, and requires the multivesicular body (MVB) pathway to release virus particles, and the released HEV particles with a lipid membrane retain the trans-Golgi network protein 2 on their surface. To examine whether HEV utilizes the exosomal pathway to release the virus particles, we analysed whether the virion release from PLC/PRF/5 cells infected with genotype 3 HEV (strain JE03-1760F) is affected by treatment with bafilomycin A1 or GW4869, or by the introduction of a small interfering RNA (siRNA) against Rab27A or Hrs. The extracellular HEV RNA titre was increased by treatment with bafilomycin A1, but was decreased by treatment with GW4869. The relative levels of virus particles released from cells depleted of Rab27A or Hrs were decreased to 16.1 and 11.5 %, respectively, of that released from cells transfected with negative control siRNA. Electron microscopic observations revealed the presence of membrane-associated virus-like particles with a diameter of approximately 50 nm within the MVB, which possessed internal vesicles in infected cells. Immunoelectron microscopy showed positive immunogold staining for the HEV ORF2 protein on the intraluminal vesicles within the MVB. Additionally, immunofluorescence analysis indicated the triple co-localization of the ORF2, ORF3 and CD63 proteins in the cytoplasm, as specific loculated signals, supporting the presence of membrane-associated HEV particles within the MVB. These findings indicate that membrane-associated HEV particles are released together with internal vesicles through MVBs by the cellular exosomal pathway.

Staining methods for sections of epon-embedded tissues for light microscopy

1970 ◽  
Vol 48 (1) ◽  
pp. 189-191 ◽  
Author(s):  
C. Roland Leeson ◽  
Thomas S. Leeson
Keyword(s):  
Light Microscopy ◽  
Synthetic Resin ◽  
Electron Microscopic ◽  
Glass Slides ◽  
Staining Methods

Sections 0.5–2 μ thick are mounted on clean glass slides and allowed to dry. A number of staining procedures are described. After the sections are stained, permanent preparations are made by mounting them in a synthetic resin. The methods result in sections which are suitable for routine light microscopy and for comparison with adjacent electron microscopic sections.

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