High-Voltage Electron Microscopy

1969 ◽  
Vol 2 (2) ◽  
pp. 95-133 ◽  
Author(s):  
V. E. Cosslett
Keyword(s):  
Electron Microscopy ◽  
High Voltage ◽  
Image Intensifier ◽  
Chromatic Aberration ◽  
Biological Tissues ◽  
Electron Gun ◽  
Resolving Power ◽  
Small Aperture ◽  
High Voltage Electron Microscopy ◽  
Voltage Electron

SummaryThe main advantage of high voltage in electron microscopy is greater penetration. When using an aperture of optimum size the thickness of specimen that can be imaged increases almost linearly with applied voltage in the case of light elements, both when the criterion is image intensity and when it is resolution. For heavy elements the increase is less rapid. With a small aperture the increase in observable thickness is still less rapid, and ‘saturates’ towards I MV. For a specimen of given thickness, image definition increases nearly linearly with voltage owing to the decrease in chromatic aberration. Although ultimate resolving power improves with voltage, the gain is slight and is offset by a fall in contrast. The optimum voltage for very high resolution is probably between 200 and 300 kV. Radiation damage arising from ionization decreases with rising voltage, making easier the examination of sensitive materials such as polymers. On the other hand, ejection of atoms by head-on collision increases rapidly above a threshold voltage, causing observable defects in metals.In construction, a high-voltage microscope differs from the normal type only in size and in having an accelerator instead of a simple electron gun. In operation it differs little, apart from precautions to avoid fiashover in the accelerator. A decrease in response of viewing screens and photographic emulsions is more than compensated by higher brightness of the electron gun. The chief applications so far of the high-voltage microscope have been for studying thick films of metals, magnetic materials, ceramics and polymers. Improved preparation techniques should make it possible to study sections of biological tissues up to 5 μ thick. The observation of micro-organisms and other specimens in the wet state can be carried out in double-walled cells, but only at poor resolution. Still higher voltages, up to 3 or MV coupled with the use of an energy analyser or an image intensifier, should improve further the microscopy of such thick specimens.

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.

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 voltage electron microscopy and its application in biology

1971 ◽  
Vol 261 (837) ◽  
pp. 35-44 ◽  
Keyword(s):  
Electron Microscopy ◽  
High Voltage ◽  
Mechanical Stability ◽  
Three Dimensional ◽  
Carbonaceous Material ◽  
Image Resolution ◽  
Resolving Power ◽  
Dimensional Structure ◽  
High Voltage Electron Microscopy ◽  
Voltage Electron

The advantages and limitations of electron microscopy at voltages up to 1 MV are outlined. Greater thickness of specimen can be examined, the increase being almost linear with applied voltage for carbonaceous material. Alternatively, a much improved image resolution is obtained from a specimen of given thickness. For such a specimen, radiation damage and temperature rise is less than at 100 kV, but these effects probably set a limit to the maximum thickness of specimen which can be examined at 1 MV . The main disadvantage is that contrast decreases with increasing voltage, as also does the response of the fluorescent screen and of photographic emulsions. The prospective slight gain in ultimate resolving power, which might make possible the imaging of atoms, is largely offset by difficulties in maintaining electrical and mechanical stability. Examples are shown of the usefulness of high voltage microscopy for examining whole chromosomes and thick sections (up to 2 µm ). Stereomicrography is necessary if the three-dimensional structure of such relatively thick specimens is to be properly evaluated. The further possibilities for investigating wet samples in special environmental cells are outlined. It is concluded that the prospects for observing living material are remote.

Comparison of Images of Crystalline Specimens by Energy-Filtering TEM at 80 keV and CTEM at 200 keV

1990 ◽  
Vol 48 (2) ◽  
pp. 62-63
Author(s):  
A. Bakenfelder ◽  
L. Reimer ◽  
R. Rennekamp
Keyword(s):  
Electron Microscopy ◽  
Energy Loss ◽  
Chromatic Aberration ◽  
Biological Tissues ◽  
Energy Window ◽  
High Voltage Electron Microscopy ◽  
Energy Filtering ◽  
Voltage Electron ◽  
Transmission Electron ◽  
Crystalline Films

One advantage of energy-filtering electron microscopy (EFEM) is to avoid the chromatic aberration of conventional transmission electron microscopy (CTEM) by the mode of electron spectroscopic imaging (ESI) using either zero-loss filtering of unscattered and elastically scattered electrons or a narrow selected energy window at the most probable loss of the electron-energy-loss spectrum (EELS). Chromatic aberration can also be reduced by high-voltage electron microscopy (HVEM). Comparisons of ESI at 80 keV and CTEM at 200 keV have already been reported for biological tissues. In this contribution we compare the imaging of evaporated crystalline films with ESI at 80 keV in a ZEISS EM902 and with CTEM at 200 keV in a Hitachi H800/NA.Zero-loss filtering at 80 keV can be applied for maximum mass-thicknesses of x=ρt≃150 μg/cm2 where the zero-loss transmission falls below 0.001 and an energy window at the most-probable energy loss can be used below ≃300 μg/cm2. Inelastic scattering preserves the Bragg contrast.

Electron Accelerators for High-Voltage Electron Microscopy (HVEM)

1972 ◽  
Vol 30 ◽  
pp. 462-463
Author(s):  
Günter O. Reinhold
Keyword(s):  
Electron Beam ◽  
High Voltage ◽  
High Energy ◽  
Electron Gun ◽  
Resolving Power ◽  
High Voltage Electron Microscopy ◽  
Voltage Electron ◽  
High Voltage Electron ◽  
Electron Microscopes ◽  
The Stability

High-voltage electron microscopes are characterized by accelerating voltages in the megavolt range (500 kV and above). Compared to conventional electron microscopes, with voltages up to 200 kV, the high-voltage instruments offer the advantage of improved resolving power (1 to 10 Å) and greater effective penetration of the electron beam. A basic difference between ordinary and high-voltage electron microscopes is the use of an electron accelerator in place of the electron gun to accelerate the electron beam to the required high energy. The resolving power of an electron microscope is determined by the stability of the accelerating voltage, the mechanical precision of the lenses and specimen stages, the stability of the lens supply current and freedom from mechanical vibrations. The symmetrical cascade generator is the only high-voltage dc power supply to meet these requirements with regard to voltage stability and vibration-free performance.

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.

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.

An Environmental Wet Cell For High Voltage Electron Microscopy

1973 ◽  
Vol 31 ◽  
pp. 18-19
Author(s):  
N.J. Tighe ◽  
H.M. Flower ◽  
P.R. Swann
Keyword(s):  
Electron Microscopy ◽  
High Temperature ◽  
Image Quality ◽  
Inelastic Scattering ◽  
High Voltage ◽  
High Voltage Electron Microscopy ◽  
Voltage Electron ◽  
Environmental Cell ◽  
Water Saturated ◽  
High Temperature Gas

A differentially pumped environmental cell has been developed for use in the AEI EM7 million volt microscope. In the initial version the column of gas traversed by the beam was 5.5mm. This permited inclusion of a tilting hot stage in the cell for investigating high temperature gas-specimen reactions. In order to examine specimens in the wet state it was found that a pressure of approximately 400 torr of water saturated helium was needed around the specimen to prevent dehydration. Inelastic scattering by the water resulted in a sharp loss of image quality. Therefore a modified cell with an ‘airgap’ of only 1.5mm has been constructed. The shorter electron path through the gas permits examination of specimens at the necessary pressure of moist helium; the specimen can still be tilted about the side entry rod axis by ±7°C to obtain stereopairs.

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.

“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.

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