Design of an intermediate high voltage EM for 3-D studies of biological material and its integration with a system for remote access

Author(s):  
Mark H. Ellisman

The increased availability of High Performance Computing and Communications (HPCC) offers scientists and students the potential for effective remote interactive use of centralized, specialized, and expensive instrumentation and computers. Examples of instruments capable of remote operation that may be usefully controlled from a distance are increasing. Some in current use include telescopes, networks of remote geophysical sensing devices and more recently, the intermediate high voltage electron microscope developed at the San Diego Microscopy and Imaging Resource (SDMIR) in La Jolla. In this presentation the imaging capabilities of a specially designed JEOL 4000EX IVEM will be described. This instrument was developed mainly to facilitate the extraction of 3-dimensional information from thick sections. In addition, progress will be described on a project now underway to develop a more advanced version of the Telemicroscopy software we previously demonstrated as a tool to for providing remote access to this IVEM (Mercurio et al., 1992; Fan et al., 1992).

Author(s):  
J.L. Williams ◽  
K. Heathcote ◽  
E.J. Greer

High Voltage Electron Microscope already offers exciting experimental possibilities to Biologists and Materials Scientists because the increased specimen thickness allows direct observation of three dimensional structure and dynamic experiments on effectively bulk specimens. This microscope is designed to give maximum accessibility and space in the specimen region for the special stages which are required. At the same time it provides an ease of operation similar to a conventional instrument.


Author(s):  
M. Nishigaki ◽  
S. Katagiri ◽  
H. Kimura ◽  
B. Tadano

The high voltage electron microscope has many advantageous features in comparison with the ordinary electron microscope. They are a higher penetrating efficiency of the electron, low chromatic aberration, high accuracy of the selected area diffraction and so on. Thus, the high voltage electron microscope becomes an indispensable instrument for the metallurgical, polymer and biological specimen studies. The application of the instrument involves today not only basic research but routine survey in the various fields. Particularly for the latter purpose, the performance, maintenance and reliability of the microscope should be same as those of commercial ones. The authors completed a 500 kV electron microscope in 1964 and a 1,000 kV one in 1966 taking these points into consideration. The construction of our 1,000 kV electron microscope is described below.


Author(s):  
Murray Vernon King ◽  
Donald F. Parsons

Effective application of the high-voltage electron microscope to a wide variety of biological studies has been restricted by the radiation sensitivity of biological systems. The problem of radiation damage has been recognized as a serious factor influencing the amount of information attainable from biological specimens in electron microscopy at conventional voltages around 100 kV. The problem proves to be even more severe at higher voltages around 1 MV. In this range, the problem is the relatively low sensitivity of the existing recording media, which entails inordinately long exposures that give rise to severe radiation damage. This low sensitivity arises from the small linear energy transfer for fast electrons. Few developable grains are created in the emulsion per electron, while most of the energy of the electrons is wasted in the film base.


Author(s):  
W. E. King

A side-entry type, helium-temperature specimen stage that has the capability of in-situ electrical-resistivity measurements has been designed and developed for use in the AEI-EM7 1200-kV electron microscope at Argonne National Laboratory. The electrical-resistivity measurements complement the high-voltage electron microscope (HVEM) to yield a unique opportunity to investigate defect production in metals by electron irradiation over a wide range of defect concentrations.A flow cryostat that uses helium gas as a coolant is employed to attain and maintain any specified temperature between 10 and 300 K. The helium gas coolant eliminates the vibrations that arise from boiling liquid helium and the temperature instabilities due to alternating heat-transfer mechanisms in the two-phase temperature regime (4.215 K). Figure 1 shows a schematic view of the liquid/gaseous helium transfer system. A liquid-gas mixture can be used for fast cooldown. The cold tip of the transfer tube is inserted coincident with the tilt axis of the specimen stage, and the end of the coolant flow tube is positioned without contact within the heat exchanger of the copper specimen block (Fig. 2).


Author(s):  
P. R. Okamoto ◽  
N.Q. Lam ◽  
R. L. Lyles

During irradiation of thin foils in a high voltage electron microscope (HVEM) defect gradients will be set up between the foil surfaces and interior. In alloys defect gradients provide additional driving forces for solute diffusion since any preferential binding and/or exchange between solute atoms and mobile defects will couple a net flux of solute atoms to the defect fluxes. Thus, during irradiation large nonequilibrium compositional gradients can be produced near the foil surfaces in initially homogeneous alloys. A system of coupled reaction-rate and diffusion equations describing the build up of mobile defects and solute redistribution in thin foils and in a semi-infinite medium under charged-particle irradiation has been formulated. Spatially uniform and nonuniform damage production rates have been used to model solute segregation under electron and ion irradiation conditions.An example calculation showing the time evolution of the solute concentration in a 2000 Å thick foil during electron irradiation is shown in Fig. 1.


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).


Author(s):  
D. Barnard ◽  
D. Rexford ◽  
W.F. Tivol ◽  
J.N. Turner

A side-entry differentially pumped environmental chamber (SEDPEC) has been designed and constructed for the AEI-EM7 high-voltage electron microscope (HVEM). The SEDPEC has been tested in the same way as previous chambers for the HVEM. In contrast to the lengthy procedures necessary to install previous environmental chambers in the HVEM, the SEDPEC can be installed in about one half hour. Thus a user can install the SEDPEC, use it for a day and return the HVEM to normal operating status without causing delays for other HVEM users. This is particularly important for our facility, which is supported as a national biotechnology resource by the NIH.


Author(s):  
David J. Smith

The initial attractions of the high voltage electron microscope (HVEM) stemmed mainly from the possibility of considerable increases in electron penetration through thick specimens compared with conventional 100KV microscopes, although the potential improvement in resolution associated with the decrease in election wavelength had been fully appreciated for many years (eg. Cosslett, 1946)1, even if not realizable in practice. Subsequent technological advances enabled the performance of lower voltage machines to be brought closer to the theoretical limit, to be followed in turn by more recent projects which have been successful, eventually, in achieving even higher resolution with dedicated higher voltage instruments such as those at Kyoto (500KV)2, Munich (400KV)3, Ibaraki (1250KV)4 and Cambridge (600KV)5. It does not necessarily follow however that the performance of journal high voltage microscopes can be easily upgraded, retrospectively, to the same level, as will be discussed in detail below.


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