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.”

Staining Serial Thick Sections For High-Voltage Electron Microscopy

1985 ◽  
Vol 43 ◽  
pp. 708-709
Author(s):  
John C. Kinnamon ◽  
Terri A. Sherman
Keyword(s):  
Electron Microscopy ◽  
High Voltage ◽  
Carbon Coating ◽  
Three Dimensional ◽  
Taste Bud ◽  
Serial Sections ◽  
High Voltage Electron Microscopy ◽  
Ultrastructural Studies ◽  
Voltage Electron ◽  
High Voltage Electron

Ultrastructural studies often require three-dimensional reconstructions from serial electron micrographs. The use of thick sections and high voltage electron microscopy can reduce the numbers of serial sections needed, but we have found that over 300 serial sections (0.5 μm thick) are still needed for our studies of taste bud ultrastructure. Serial reconstructions are almost always done with sections mounted onto formvar-coated slot grids, because they provide an unobscured view of the specimen. Staining and carbon coating several hundred formvar-coated slot grids often results in the loss of serial sections because the formvar films break due to handling during the staining and carbon-coating procedures. In addition, the staining process itself can be tedious and time-consuming when staining thick sections because staining times of hours are often required. Such lengthy staining times may result in staining artifacts due to exposure of the stain to the atmosphere, especially with lead stains.

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.

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.

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.

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