Application of Imaging Plate to High-Voltage Electron Microscopy

Author(s):  
Seiji Isoda ◽  
Kimitsugu Saitoh ◽  
Sakumi Moriguchi ◽  
Takashi Kobayashi

On the observation of structures by high resolution electron microscopy, recording materials with high sensitivity and high quality is awaited, especially for the study of radiation sensitive specimens. Such recording material should be easily combined with the minimum dose system and cryoprotection method. Recently a new recording material, imaging plate, comes to be widely used in X-ray radiography and also in electron microscopy, because of its high sensitivity, high quality and the easiness in handling the images with a computer. The properties of the imaging plate in 100 to 400 kV electron microscopes were already discussed and the effectiveness was revealed.It is demanded to study the applicability of the imaging plate to high voltage electron microscopy. The quality of the imaging plate was investigated using an imaging plate system (JEOL EM-HSR100) equipped in a new Kyoto 1000kV electron microscope. In the system both the imaging plate and films can be introduced together into the camera chamber. Figure 1 shows the effect of accelerating voltage on read-out signal intensities from the imaging plate. The characteristic of commercially available imaging plates is unfortunately optimized for 100 to 200 keV electrons and then for 600 to 1000 keV electrons the signal is reduced. In the electron dose range of 10−13 to 10−10 C/cm2, the signal increases linearly with logarithm of electron dose in all acceralating volatges.

Author(s):  
L. D. Ackerman ◽  
S. H. Y. Wei

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.


Author(s):  
Lee D. Peachey ◽  
Clara Franzini-Armstrong

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.


Author(s):  
T. Mukai ◽  
T. E. Mitchell

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.


Author(s):  
G. E. Tyson ◽  
M. J. Song

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.


Author(s):  
William H. Massover

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.


Author(s):  
Benjamin M. Siegel

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.


Author(s):  
J.N. Turner ◽  
W.G. Shain ◽  
V. Madelian ◽  
R.A. Grassucci ◽  
D.L. Forman

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.


Author(s):  
Conly L. Rieder ◽  
Frederick J. Miller ◽  
Edwin Davison ◽  
Samuel S. Bowser ◽  
Kirsten Lewis ◽  
...  

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.


Author(s):  
Charles W. Allen

High voltage TEMs were introduced commercially thirty years ago, with the installations of 500 kV Hitachi instruments at the Universities of Nogoya and Tokyo. Since that time a total of 51 commercial instruments, having maximum accelerating potentials of 0.5-3.5 MV, have been delivered. Prices have gone from about a dollar per volt for the early instruments to roughly twenty dollars per volt today, which is not so unreasonable considerinp inflation and vastly improved electronics and other improvements. The most expensive HVEM (the 3.5 MV instrument at Osaka University) cost about 5 percent of the construction cost of the USA's latest synchrotron.Table 1 briefly traces the development of HVEM in this country for the materials sciences. There are now only three available instruments at two sites: the 1.2 MeV HVEM at Argonne National Lab, and 1.0 and 1.5 MeV instruments at Lawrence Berkeley National Lab. Fortunately, both sites are user facilities funded by DOE for the materials research community.


Author(s):  
Brenda R. Eisenberg ◽  
Lee D. Peachey

Analysis of the electrical properties of the t-system requires knowledge of the geometry of the t-system network. It is now possible to determine the network parameters experimentally by use of high voltage electron microscopy. The t-system was marked with exogenous peroxidase. Conventional methods of electron microscopy were used to fix and embed the sartorius muscle from four frogs. Transverse slices 0.5-1.0 μm thick were viewed at an accelerating voltage of 1000 kV using the JEM-1000 high voltage electron microscope at Boulder, Colorado and prints at x5000 were used for analysis.The length of a t-branch (t) from node to node (Fig. 1a) was measured with a magnifier; at least 150 t-branches around 30 myofibrils were measured from each frog. The mean length of t is 0.90 ± 0.11 μm and the number of branches per myofibril is 5.4 ± 0.2 (mean ± SD, n = 4 frogs).


Sign in / Sign up

Export Citation Format

Share Document