Ultrastructural comparisons of fungus hyphal cells using frozen-etched replicas and thin sections of the fungus Pyrenochaeta terrestris

1968 ◽  
Vol 14 (3) ◽  
pp. 205-210 ◽  
Author(s):  
W. M. Hess
Keyword(s):  
Electron Scattering ◽  
Cell Walls ◽  
Sole Carbon Source ◽  
Membrane Systems ◽  
Thin Sections ◽  
Fixed Membrane ◽  
Pyrenochaeta Terrestris ◽  
Freeze Etching ◽  
Fixed Cell ◽  
Etched Membrane

The ultrastructure of P. terrestris hyphal cells was investigated to compare frozen-etched replicas with chemically fixed thin sections. The fungus used in this study uses glycerol as a sole carbon source and survives the freezing procedures necessary for freeze-etching; thus frozen-etched replicas reflect the living state.Frozen-etched membrane systems have particles of various sizes and concentrations and have a smooth appearance as contrasted to chemically fixed membrane systems, which have particles difficult to distinguish and somewhat irregular membrane systems. Frozen-etched cell walls are seen to contain particles, and microfibrillar orientation is evident in older cell walls, whereas substructure is not evident in chemically fixed cell walls, although secretion products of the fungus accumulate on cell surfaces.Chemically fixed ground cytoplasm has ribosomes and areas of high- and low-electron scattering which are not seen with freeze-etching. Cells fixed in glutaraldehyde–acrolein–OsO4 more nearly resemble frozen-etched cells than cells fixed in potassium permanganate.

Ultrastructure of Staphylococcus epidermidis after freeze-etching and thin sectioning

1973 ◽  
Vol 19 (2) ◽  
pp. 294-295
Author(s):  
James E. Gilchrist ◽  
Irving W. DeVoe
Keyword(s):  
Staphylococcus Aureus ◽  
Outer Layer ◽  
Staphylococcus Epidermidis ◽  
Cell Walls ◽  
Cytoplasmic Membrane ◽  
Middle Layer ◽  
Dense Layer ◽  
Thin Sections ◽  
Thin Sectioning ◽  
Freeze Etching

A considerable quantity of information is now available on the ultrastructure of Staphylococcus (1, 2, 4, 7, 8, 10, 11, 12). Cell walls of these organisms in thin sections have been shown to consist of three layers: a dense outer layer, a rather electron translucent middle layer, and a very dense layer next to the cytoplasmic membrane (2, 7, 11, 12). Bulger and Bulger (2) have pointed out the presence of circumferential substructure in the middle layer of the wall in a strain of Staphylococcus aureus isolated as the causative agent in fatal pneumonia.Numerous mesosomes of both the vesicular and laminar types are evident in thin sections of staphylococci from several studies (1, 4, 7, 11). Moreover, single vesicular structures that appear to be invaginations of the trilaminar cytoplasmic membrane have been pointed out by Suganuma (11) and Beaton (1).

The response of cell walls of Bacillus subtilis to metals and to electron-microscopic stains

1978 ◽  
Vol 24 (2) ◽  
pp. 89-104 ◽  
Author(s):  
T. J. Beveridge
Keyword(s):  
Bacillus Subtilis ◽  
Electron Scattering ◽  
Cell Walls ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Ruthenium Red ◽  
Transition Elements ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Wall Ultrastructure

Purified cell walls of Bacillus subtilis were subjected to solutions of 40 independent metals and the metal uptake, the electron-scattering power of thin sections, and the type of staining response evaluated. This was repeated for six typical electron-microscopic stains (uranyl acetate, uranyl magnesium acetate, osmium tetroxide, Os-meth, osmium-dimethylethylenediamine, and ruthenium red) and one new staining reagent (a potassium platinum chloride – dimethylsulfoxide complex) whose specificity is for amine functions. The reaction of select metals can be specific in terms of both uptake and staining response. Of the metals studied most transition elements had a high affinity for the wall fabric and some (i.e., Sc III, most lanthanides, U IV, Zr IV, Hf IV, Fe III, Pd II, Ru III, and In III) may be suitable as contrasting agents for electron microscopy. Furthermore, when the thickness of metal-reacted walls was compared to freeze-each and ultracryotomy data, statistical-dimensional differences were commonly seen, which indicates that wall ultrastructure can be profoundly affected by the type of metal and (or) staining reagent.

Ectodesmata as Revealed By Freeze-Etch Preparations of Plant Epidermal Cell Walls

1973 ◽  
Vol 31 ◽  
pp. 468-469
Author(s):  
N.C. Lyon ◽  
W. C. Mueller
Keyword(s):  
Electron Microscopy ◽  
Acetic Acid ◽  
Nitric Acid ◽  
Oxalic Acid ◽  
Light Microscopy ◽  
Epidermal Cell ◽  
Cell Walls ◽  
Plant Viruses ◽  
Plant Leaves ◽  
Thin Sections

Schumacher and Halbsguth first demonstrated ectodesmata as pores or channels in the epidermal cell walls in haustoria of Cuscuta odorata L. by light microscopy in tissues fixed in a sublimate fixative (30% ethyl alcohol, 30 ml:glacial acetic acid, 10 ml: 65% nitric acid, 1 ml: 40% formaldehyde, 5 ml: oxalic acid, 2 g: mecuric chloride to saturation 2-3 g). Other workers have published electron micrographs of structures transversing the outer epidermal cell in thin sections of plant leaves that have been interpreted as ectodesmata. Such structures are evident following treatment with Hg++ or Ag+ salts and are only rarely observed by electron microscopy. If ectodesmata exist without such treatment, and are not artefacts, they would afford natural pathways of entry for applied foliar solutions and plant viruses.

Chemical modification of the bacterial wall to determine sites of metal deposition

1978 ◽  
Vol 36 (2) ◽  
pp. 350-351
Author(s):  
T. J. Beveridge
Keyword(s):  
Electron Scattering ◽  
Metal Salt ◽  
Metal Deposition ◽  
Suitable Model ◽  
X Ray Diffraction ◽  
Subtilis Cell ◽  
Thin Sections ◽  
X Ray ◽  
Section Data ◽  
Metal Impregnation

The Bacillus subtilis cell wall provides a protective sacculus about the vital constituents of the bacterium and consists of a collection of anionic hetero- and homopolymers which are mainly polysaccharidic. We recently demonstrated that unfixed walls were able to trap and retain substantial amounts of metal when suspended in aqueous metal salt solutions. These walls were briefly mixed with low concentration metal solutions (5mM for 10 min at 22°C), were well washed with deionized distilled water, and the quantity of metal uptake (atomic absorption and X-ray fluorescence), the type of staining response (electron scattering profile of thin-sections), and the crystallinity of the deposition product (X-ray diffraction of embedded specimens) determined.Since most biological material possesses little electron scattering ability electron microscopists have been forced to depend on heavy metal impregnation of the specimen before obtaining thin-section data. Our experience with these walls suggested that they may provide a suitable model system with which to study the sites of reaction for this metal deposition.

Quantitation in thin Sections Within the Electron Microscope

1976 ◽  
Vol 34 ◽  
pp. 336-337
Author(s):  
J. Edie
Keyword(s):  
Electron Beam ◽  
Electron Microscope ◽  
Electron Scattering ◽  
Volume Density ◽  
Thin Sections ◽  
Faraday Cage ◽  
Local Mass ◽  
Physical Measurements ◽  
Mass Volume ◽  
Varying Mass

In TEM image formation, the observed contrast variations within thin sections result from differential electron scattering within microregions of varying mass thickness. It is possible to utilize these electron scattering properties to obtain objective information regarding various specimen parameters (1, 2, 3).A pragmatic, empirical approach is described which enables a microscopist to perform physical measurements of thickness of thin sections and estimates of local mass, volume, density and, possibly, molecular configurations within thin sections directly in the microscope. A Faraday cage monitors the transmitted electron beam and permits measurements of electron beam intensities.

scholarly journals FINE STRUCTURE OF THE GRAM-NEGATIVE BACTERIUM ACETOBACTER SUBOXYDANS

1964 ◽  
Vol 20 (2) ◽  
pp. 217-233 ◽  
Author(s):  
G. W. Claus ◽  
L. E. Roth
Keyword(s):  
Cell Wall ◽  
Plasma Membrane ◽  
Cell Walls ◽  
Negative Staining ◽  
Homogeneous Layer ◽  
Physical Treatment ◽  
Thin Sections ◽  
Gram Negative ◽  
Acetobacter Suboxydans ◽  
Negative Cell

The morphological features of the cell wall, plasma membrane, protoplasmic constituents, and flagella of Acetobacter suboxydans (ATCC 621) were studied by thin sectioning and negative staining. Thin sections of the cell wall demonstrate an outer membrane and an inner, more homogeneous layer. These observations are consistent with those of isolated, gram-negative cell-wall ghosts and the chemical analyses of gram-negative cell walls. Certain functional attributes of the cell-wall inner layer and the structural comparisons of gram-negative and gram-positive cell walls are considered. The plasma membrane is similar in appearance to the membrane of the cell wall and is occasionally found to be folded into the cytoplasm. Certain features of the protoplasm are described and discussed, including the diffuse states of the chromatinic material that appear to be correlated with the length of the cell and a polar differentiation in the area of expected flagellar attachment. Although the flagella appear hollow in thin sections, negative staining of isolated flagella does not substantiate this finding. Severe physical treatment occasionally produces a localized penetration into the central region of the flagellum, the diameter of which is much smaller then that expected from sections. A possible explanation of this apparent discrepancy is discussed.

The Structure of the Macromolecular Units on the Cell Walls of Bacillus Polymyxa

1967 ◽  
Vol 2 (4) ◽  
pp. 587-591
Author(s):  
J. T. FINCH ◽  
A. KLUG ◽  
M.V. NERMUT
Keyword(s):  
Cell Wall ◽  
Electron Micrographs ◽  
Electron Scattering ◽  
Cell Walls ◽  
Single Layer ◽  
Optical Diffraction ◽  
Square Lattice ◽  
Bacillus Polymyxa ◽  
Optical Filtering ◽  
Diffraction Patterns

Electron micrographs of negatively stained preparations of cell walls of Bacillus polymyxa have been investigated by optical diffraction and optical filtering techniques. Images of single layers of the cell wall, from which the ‘noise’ has been filtered optically, show hollow, square-shaped morphological units arranged on a square lattice of side 100 Å. Single-layer images showing the same pattern have been filtered from moiré patterns arising from two overlapping single layers. The morphological units are composed of four smaller subunits. The optical diffraction patterns from regions of two overlapping layers show extra reflexions which are attributed to multiple electron scattering.

Effects of Hormones on the Polysaccharide-Synthesizing Membrane Systems of Lettuce Pith

1974 ◽  
Vol 16 (2) ◽  
pp. 433-443
Author(s):  
K. WRIGHT ◽  
D. J. BOWLES
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Walls ◽  
Membrane Systems ◽  
Radioactive Precursor

Changes have been induced in the polysaccharides laid down in the cell walls of lettuce pith by administration of the hormones auxin and zeatin. This polysaccharide derives from the membrane systems of the cells and radioactive precursor has been used to follow the accompanying changes which occurred in the polysaccharide contained within isolated Golgi and endoplasmic reticulum fractions. Zeatin-induced division and differentiation was accompanied by up to 12-fold increases in the amount of radioactivity in polysaccharide of isolated membrane fractions, and the pattern of distribution of label between the sugars of this polysaccharide was qualitatively different in the presence of zeatin. The change in this pattern was evident in the Golgi fraction at an earlier stage in the induction of the response than that in the endoplasmic reticulum.

LYSOZYME SENSITIVITY OF THE CELL WALL OF PSEUDO-MONAS AERUGINOSA: FURTHER EVIDENCE FOR THE ROLE OF THE NON-PEPTIDOGLYCAN COMPONENTS IN CELL WALL RIGIDITY

1966 ◽  
Vol 12 (1) ◽  
pp. 105-108 ◽  
Author(s):  
K. Jane Carson ◽  
R. G. Eagon
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Cell Wall ◽  
Electron Micrographs ◽  
Cell Walls ◽  
Gram Negative Bacteria ◽  
Normal Cells ◽  
Thin Sections ◽  
Gram Negative ◽  
Cell Wall Rigidity

Electron micrographs of thin sections of normal cells of Pseudomonas aeruginosa showed the cell walls to be convoluted and to be composed of two distinct layers. Electron micrographs of thin sections of lysozyme-treated cells of P. aeruginosa showed (a) that the cell walls lost much of their convoluted nature; (b) that the layers of the cell walls became diffuse and less distinct; and (c) that the cell walls became separated from the protoplasts over extensive cellular areas. These results suggest that the peptidoglycan component of the unaltered cell walls of P. aeruginosa is sensitive to lysozyme. Furthermore, it appears that the peptidoglycan component is not solely responsible for the rigidity of the cell walls of Gram-negative bacteria.

Fine structure of the thermophilic blue-green alga Synechococcus lividus Copeland. A study of frozen–fractured–etched cells

1972 ◽  
Vol 18 (2) ◽  
pp. 175-181 ◽  
Author(s):  
S. C. Holt ◽  
M. R. Edwards
Keyword(s):  
Green Alga ◽  
Lipid Bodies ◽  
Blue Green Alga ◽  
Etching Technique ◽  
Thin Sections ◽  
Blue Green Algae ◽  
Osmiophilic Granules ◽  
Freeze Etching ◽  
The Many ◽  
Synechococcus Lividus

The thermophilic unicellular blue-green alga, Synechococcus lividus, was studied by electron microscopy in thin sections and by the freeze-etching technique. Thin sections revealed subcellular structures like those observed by other authors in mesophilic blue-green algae. In the freeze-etched fractures similar results were obtained but, in addition, surface views of plasma and thylakoidal membranes were examined in detail. The many inclusions present in the freeze-etched preparations confirmed those displayed in thin sections and are interpreted as polyhedral, polyphosphate, and lipid bodies. Some unidentified osmiophilic granules and also phycobilisomes were seen.

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