Some Biological Applications of High Voltage Electron Microscopy

1974 ◽  
Vol 32 ◽  
pp. 298-299
Author(s):  
Hans Ris
Keyword(s):  
High Voltage ◽  
Chromosome Structure ◽  
Nerve Fibers ◽  
Good Resolution ◽  
High Voltage Electron Microscopy ◽  
Voltage Electron ◽  
University Of Wisconsin ◽  
High Voltage Electron ◽  
One Year ◽  
The University

The High Voltage Electron Microscope Laboratory at the University of Wisconsin has been in operation a little over one year. I would like to give a progress report about our experience with this new technique. The achievement of good resolution with thick specimens has been mainly exploited so far. A cold stage which will allow us to look at frozen specimens and a hydration stage are now being installed in our microscope. This will soon make it possible to study undehydrated specimens, a particularly exciting application of the high voltage microscope.Some of the problems studied at the Madison facility are: Structure of kinetoplast and flagella in trypanosomes (J. Paulin, U. of Georgia); growth cones of nerve fibers (R. Hannah, U. of Georgia Medical School); spiny dendrites in cerebellum of mouse (Scott and Guillery, Anatomy, U. of Wis.); spindle of baker's yeast (Joan Peterson, Madison) spindle of Haemanthus (A. Bajer, U. of Oregon, Eugene) chromosome structure (Hans Ris, U. of Wisconsin, Madison). Dr. Paulin and Dr. Hanna are reporting their work separately at this meeting and I shall therefore not discuss it here.

Integrated microscopy resource for biomedical research at the university of wisconsin at madison

1987 ◽  
Vol 45 ◽  
pp. 594-597 ◽  
Author(s):  
G. Schatten ◽  
J. Pawley ◽  
H. Ris
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Transmission Electron Microscope ◽  
High Voltage ◽  
Biomedical Research ◽  
High Voltage Electron Microscopy ◽  
Voltage Electron ◽  
University Of Wisconsin ◽  
Transmission Electron ◽  
The University

The High Voltage Electron Microscopy Laboratory [HVEM] at the University of Wisconsin-Madison, a National Institutes of Health Biomedical Research Technology Resource, has recently been renamed the Integrated Microscopy Resource for Biomedical Research [IMR]. This change is designed to highlight both our increasing abilities to provide sophisticated microscopes for biomedical investigators, and the expansion of our mission beyond furnishing access to a million-volt transmission electron microscope. This abstract will describe the current status of the IMR, some preliminary results, our upcoming plans, and the current procedures for applying for microscope time.The IMR has five principal facilities: 1.High Voltage Electron Microscopy2.Computer-Based Motion Analysis3.Low Voltage High-Resolution Scanning Electron Microscopy4.Tandem Scanning Reflected Light Microscopy5.Computer-Enhanced Video MicroscopyThe IMR houses an AEI-EM7 one million-volt transmission electron microscope.

High-voltage electron microscopy and 3-D reconstruction of solitary chemosensory cells and Di-I labeling of primary afferent nerves

1990 ◽  
Vol 48 (3) ◽  
pp. 412-413
Author(s):  
Kurt Kotrschal ◽  
John C. Kinnamon ◽  
Suzanne M. Royer
Keyword(s):  
Electron Microscopy ◽  
High Voltage ◽  
Nerve Fibers ◽  
Taste Bud ◽  
Computer Assisted ◽  
High Voltage Electron Microscopy ◽  
Voltage Electron ◽  
High Voltage Electron ◽  
Solitary Chemosensory Cells ◽  
Taste Bud Cells

Solitary chemosensory cells (SCCs) resembling taste bud cells in shape and ultrastructure have been found in fish and amphibians. A quantitative survey in teleosts has revealed that SCCs may be even more abundant than taste bud cells. Due to their scattered distribution, however, this abundant and potentially important vertebrate chemosensory system has eluded closer examination. This situation has changed with the introduction of a suitable research model: the anterior dorsal fin (ADF) of rocklings (Gadidae, Teleostei). This specialized chemosensory organ contains several million SCCs that converge onto fine caliber recurrent facial nerve fibers. The ADF model has allowed us to investigate SCC innervation, fine structure and function.Using high voltage electron microscopy of serial sections and computer-assisted three-dimensional reconstructions, we have obtained significant quantitative data on the spatial structure of SCC assemblages in the ADF epidermis and of synaptic contacts between SCCs and innervating facial nerves. Approximately 15% of all cells present in the ADF epidermis are SCCs, making up 30% of the epidermal volume. Unlike taste buds, the ADF contains only a single sensory cell type and lacks basal cells or specialized “supporting cells.”

High Voltage Electron Microscopy of Ion-Milled Dental Enamel

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.

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

1977 ◽  
Vol 35 ◽  
pp. 570-571 ◽  
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.

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

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.

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

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.

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

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.

“High Resolution High Voltage Electron Microscopy”

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.

High-voltage Electron Microscopy of astroglial cell whole mounts

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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