Colloidal Gold in High-Voltage Electron Microscopy–Ruthenium Red Method and Whole-Cell Mount

One of the advantages of high voltage electron microscopy (HVEM) is the great penetration of electron beam, which permits the examination of thicker sections of resin-embedded biological materials. However, when stained thick sections are examined under the HVEM, all structural details within the whole thickness are brought into focus as superimposed image. Hence, certain structures are not easily observable on such an image, even on the stereo view obtained by specimen tilting technique. To overcome this difficulty, molecular tracers are introduced to the HVEM in an attempt to visualize certain structures selectively.Mouse kidney and cardiac muscle were examined using ruthenium red and peroxidase as molecular tracers. Thick Epon sections ranging from 0.5 to 3 μ were observed in a JOEL JEM-1000 or a Hitachi HU-1000 HVEM, with an accelerating voltage of 1000 KV. Pictures were taken routinely as stereo pairs using a tilting stage device with ±10° or ±8°.

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High Voltage Electron Microscopy of Ion-Milled Dental Enamel

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100050317 ◽

1974 ◽

Vol 32 ◽

pp. 60-61

Author(s):

L. D. Ackerman ◽

S. H. Y. Wei

Keyword(s):

Electron Microscopy ◽

High Voltage ◽

Third Molar ◽

Polarized Light ◽

Dental Enamel ◽

Longitudinal Section ◽

Ion Milling ◽

High Voltage Electron Microscopy ◽

Voltage Electron ◽

High Voltage Electron

Mature human dental enamel has presented investigators with several difficulties in ultramicrotomy of specimens for electron microscopy due to its high degree of mineralization. This study explores the possibility of combining ion-milling and high voltage electron microscopy as a means of circumventing the problems of ultramicrotomy.A longitudinal section of an extracted human third molar was ground to a thickness of about 30 um and polarized light micrographs were taken. The specimen was attached to a single hole grid and thinned by argon-ion bombardment at 15° incidence while rotating at 15 rpm. The beam current in each of two guns was 50 μA with an accelerating voltage of 4 kV. A 20 nm carbon coating was evaporated onto the specimen to prevent an electron charge from building up during electron microscopy.

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Three-Dimensional Visualization of the T-System of Frog Muscle Using High Voltage Electron Microscopy and a Lanthanum Stain

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100080286 ◽

1977 ◽

Vol 35 ◽

pp. 570-571 ◽

Cited By ~ 2

Author(s):

Lee D. Peachey ◽

Clara Franzini-Armstrong

Keyword(s):

Electron Microscopy ◽

High Voltage ◽

Three Dimensional ◽

Biological Tissues ◽

High Voltage Electron Microscopy ◽

Transverse Tubular System ◽

Peroxidase Reaction ◽

Voltage Electron ◽

High Voltage Electron ◽

T System

The effective study of biological tissues in thick slices of embedded material by high voltage electron microscopy (HVEM) requires highly selective staining of those structures to be visualized so that they are not hidden or obscured by other structures in the image. A tilt pair of micrographs with subsequent stereoscopic viewing can be an important aid in three-dimensional visualization of these images, once an appropriate stain has been found. The peroxidase reaction has been used for this purpose in visualizing the T-system (transverse tubular system) of frog skeletal muscle by HVEM (1). We have found infiltration with lanthanum hydroxide to be particularly useful for three-dimensional visualization of certain aspects of the structure of the T- system in skeletal muscles of the frog. Specifically, lanthanum more completely fills the lumen of the tubules and is denser than the peroxidase reaction product.

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Radiation-Induced Precipitation at Thin Foil Surfaces in Ni-Be Alloys

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100055217 ◽

1982 ◽

Vol 40 ◽

pp. 598-599

Author(s):

T. Mukai ◽

T. E. Mitchell

Keyword(s):

Electron Microscopy ◽

High Voltage ◽

Electron Irradiation ◽

High Vacuum ◽

Thin Foil ◽

Homogeneous Precipitation ◽

High Voltage Electron Microscopy ◽

Voltage Electron ◽

High Voltage Electron ◽

Radiation Induced

Radiation-induced homogeneous precipitation in Ni-Be alloys was recently observed by high voltage electron microscopy. A coupling of interstitial flux with solute Be atoms is responsible for the precipitation. The present investigation further shows that precipitation is also induced at thin foil surfaces by electron irradiation under a high vacuum.

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High-voltage electron microscopy of maxillary gland of spirochete-infected brine shrimp: lesion of efferent tubule

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100103875 ◽

1988 ◽

Vol 46 ◽

pp. 362-363

Author(s):

G. E. Tyson ◽

M. J. Song

Keyword(s):

Electron Microscopy ◽

High Voltage ◽

Brine Shrimp ◽

Natural Populations ◽

Three Dimensional ◽

Dimensional Structure ◽

High Voltage Electron Microscopy ◽

Voltage Electron ◽

High Voltage Electron ◽

Infected Adult

Natural populations of the brine shrimp, Artemia, may possess spirochete- infected animals in low numbers. The ultrastructure of Artemia's spirochete has been described by conventional transmission electron microscopy. In infected shrimp, spirochetal cells were abundant in the blood and also occurred intra- and extracellularly in the three organs examined, i.e. the maxillary gland (segmental excretory organ), the integument, and certain muscles The efferent-tubule region of the maxillary gland possessed a distinctive lesion comprised of a group of spirochetes, together with numerous small vesicles, situated in a cave-like indentation of the base of the tubule epithelium. in some instances the basal lamina at a lesion site was clearly discontinuous. High-voltage electron microscopy has now been used to study lesions of the efferent tubule, with the aim of understanding better their three-dimensional structure.Tissue from one maxillary gland of an infected, adult, female brine shrimp was used for HVEM study.

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On the High Voltage Electron Microscopy (1 MeV) of Biological Thick Sections

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100095571 ◽

1972 ◽

Vol 30 ◽

pp. 464-465

Author(s):

William H. Massover

Keyword(s):

Electron Microscopy ◽

High Voltage ◽

Present Report ◽

Biological Tissues ◽

Specimen Preparation ◽

Technical Problems ◽

High Voltage Electron Microscopy ◽

Thin Sections ◽

Voltage Electron ◽

High Voltage Electron

Stereoscopic examination of thick sections of fixed and embedded biological tissues by high voltage electron microscopy has been shown to allow direct visualization of three-dimensional fine structure. The present report will consider the occurrence of some new technical problems in specimen preparation and Image interpretation that are not common during lower voltage studies of thin sections.Thick Sectioning and Tissue Coloration - Epon sections of 0.5 μm or more that are cut with glass knives do not have a uniform thickness as Judged by their interference colors; these colors change with time during their flotation on the knife bath, and again when drying onto the specimen support. Quoted thicknesses thus must be considered only as rough estimates unless measured in specific regions by other methods. Chloroform vapors do not always result in good spreading of thick sections; however, they will spread spontaneously to large degrees after resting on the flotation bath for several minutes. Ribbons of thick sections have been almost impossible to obtain.

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“High Resolution High Voltage Electron Microscopy”

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100101372 ◽

1968 ◽

Vol 26 ◽

pp. 326-327

Author(s):

Benjamin M. Siegel

Keyword(s):

Electron Microscopy ◽

High Resolution ◽

High Voltage ◽

Full Potential ◽

Contrast Imaging ◽

High Voltage Electron Microscopy ◽

Low Contrast ◽

Voltage Electron ◽

Critical Areas ◽

High Voltage Electron

The potential advantages of high voltage electron microscopy for extending the limits of resolution and contrast in imaging low contrast objects, such as biomolecular specimens, is very great. The results of computations will be presented showing that at accelerating voltages of 500-1000 kV it should be possible to achieve spacial resolutions of 1 to 1.5 Å and using phase contrast imaging achieve adequate image contrast to observe single atoms of low atomic number.The practical problems associated with the design and utilization of the high voltage instrument are, optimistically, within the range of competence of the state of the art. However, there are some extremely important and critical areas to be systematically investigated before we have achieved this competence. The basic electron optics of the column required is well understood, but before the full potential of an instrument capable of resolutions of better than 1.5 Å are realized some very careful development work will be required. Of great importance for the actual achievement of high resolution with a high voltage electron microscope is the fundamental limitation set by the characteristics of the high voltage electron beam that can be obtained from the accelerator column.

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High-voltage Electron Microscopy of astroglial cell whole mounts

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100143201 ◽

1986 ◽

Vol 44 ◽

pp. 316-317

Author(s):

J.N. Turner ◽

W.G. Shain ◽

V. Madelian ◽

R.A. Grassucci ◽

D.L. Forman

Keyword(s):

Electron Microscopy ◽

High Voltage ◽

Time Lapse ◽

Astroglial Cells ◽

High Voltage Electron Microscopy ◽

Rat Glioma ◽

Voltage Electron ◽

High Voltage Electron ◽

Nervous System Function ◽

Beta Adrenergic Agonist

Homogeneous cultures of astroglial cells have proved useful for studying biochemical, pharmacological, and toxicological responses of astrocytes to effectors of central nervous system function. LRM 55 astroglial cells, which were derived from a rat glioma and maintained in continuous culture, exhibit a number of astrocyte properties (1-3). Stimulation of LRM 55s and astrocytes in primary cell cultures with the beta-adrenergic agonist isoproterenol results in rapid changes of morphology. Studies with time lapse video light microscopy (VLM) and high-voltage electron microscopy (HVEM) have been correlated to changes in intracellular levels of c-AMP. This report emphasizes the HVEM results.

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Quantitative ultrastructural reconstruction of small regions within large volumes by correlative light and high voltage electron microscopy of serial 0.25-0.50 μm sections

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100127396 ◽

1987 ◽

Vol 45 ◽

pp. 578-581

Author(s):

Conly L. Rieder ◽

Frederick J. Miller ◽

Edwin Davison ◽

Samuel S. Bowser ◽

Kirsten Lewis ◽

...

Keyword(s):

Electron Microscopy ◽

High Voltage ◽

Mitotic Spindle ◽

Meiotic Spindle ◽

High Voltage Electron Microscopy ◽

Male And Female ◽

Voltage Electron ◽

High Voltage Electron ◽

Differential Stability ◽

Sperm Aster

In this abstract we Illustrate how same-section correlative light and high voltage electron microscopy (HVEM) of serial 0.25-0.50-μm sections can answer questions which are difficult to approach by EM of 60-100 nm sections.Starfish (Pisaster and Asterlas) eggs are fertilized at meiosis I when the oocyte contains two maternal centrosomes (e.g., asters) which form the poles of the first meiotic spindle. Immediately after fertilization a sperm aster is assembled in the vicinity of the male pronucleus and persists throughout meiosis. At syngamy the sperm aster splits to form the poles of the first mitotic spindle. During this time the functional and replicative properties of the maternal centrosome, inherited from the last meiotic division, are lost. The basis for this differential stability, of male and female centrosomes in the same cytoplasm, is a mystery.

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