Platinum shadowing, complementary imaging and stereo analysis: Techniques developed for TEM can now be used to increase the resolution of frozen hydrated specimens in a SEM

Author(s):  
William P. Wergin ◽  
Eric F. Erbe

In 1957, Steere froze virus crystals, fractured and transferred them to a vacuum evaporator where they were etched, shadowed and coated. The coating, which was then removed from the tissue and mounted on a grid, represented a replica that could be observed in the transmission electron microscope (TEM). This accomplishment is regarded as the first successful biological application of the freeze-etching technique in electron microscopy. This technique was further advanced by describing procedures whereby the replicas from both surfaces of a frozen, fractured sample could be recovered. This pair of replicas provided additional surface information by allowing one to view and compare the two opposing surfaces, or the complementary images, of a freeze-fractured biological specimen. Using a goniometer stage the replicas were also photographed, tilted and rephotographed to produce stereo pairs that could be used to analyze three-dimensionally the surface characteristics of the replica. Until recently these TEM techniques have been the principle procedures utilized by biologists to gain high resolution structural details about virus crystals, biological membranes and a wide range of other material.

The freeze-etching technique must be improved if structures at the molecular size level are to be seen. The limitations of the technique are discussed here together with the progress made in alleviating them. The vitrification of living specimens is limited by the fact that very high freezing rates are needed. The critical freezing rate can be lowered on the one hand by the introduction of antifreeze agents, on the other hand by the application of high hydrostatic pressure. The fracture process may cause structural distortions in the fracture face of the frozen specimen. The ‘double-replica’ method allows one to evaluate such artefacts and provides an insight into the way that membranes split. During etching there exists the danger of contaminating the fracture faces with condensable gases. Because of specimen temperatures below —110 °C, special care has to be taken in eliminating water vapour from the high vacuum. An improvement in coating freeze-etched specimens has resulted from the application of electron guns for evaporation of the highest melting-point metals. If heat transfer from gun to specimen is reduced to a minimum, Pt, Ir, Ta, W and C can be used for shadow casting. Best results are obtained with Pt-C and Ta-W . With the help of decoration effects Pt-C shadow castings give the most information about the fine structural details of the specimen.


1966 ◽  
Vol 29 (3) ◽  
pp. 435-447 ◽  
Author(s):  
Stanley Bullivant ◽  
Adelbert Ames

A simple method to achieve results similar to the freeze-etching technique of Moor et al. (1961) is described. The frozen tissue is cut under liquid nitrogen with a razor blade outside the evaporator rather than inside with a cooled microtome. The conditions of the experiment do not favor sublimation, and it is proposed that the structure of the replica be explained by local faults in the cleavage plane which leaves structures, such as membranes, standing above the ice. Micrographs of replicas of glycerol-protected frozen small intestine of mouse prepared by the method are presented and the structural details they show are discussed. The problem of vapor-deposited contamination is discussed. It is concluded that this is a practical method for obtaining electron micrographs that are relatively free of artifact, and that further improvements may be expected from the use of rapidly frozen fresh tissue and a clean vacuum system, possibly of the ion-pumped type.


Author(s):  
G. G. Cocks ◽  
C. E. Cluthe

The freeze etching technique is potentially useful for examining dilute solutions or suspensions of macromolecular materials. Quick freezing of aqueous solutions in Freon or propane at or near liquid nitrogen temperature produces relatively large ice crystals and these crystals may damage the structures to be examined. Cryoprotective agents may reduce damage to the specimem, hut their use often results in the formation of a different set of specimem artifacts.In a study of the structure of polyethylene oxide gels glycerol and sucrose were used as cryoprotective agents. The experiments reported here show some of the structures which can appear when these cryoprotective agents are used.Figure 1 shows a fractured surface of a frozen 25% aqueous solution of sucrose. The branches of dendritic ice crystals surrounded hy ice-sucrose eutectic can be seen. When this fractured surface is etched the ice in the dendrites sublimes giving the type of structure shown in Figure 2. The ice-sucrose eutectic etches much more slowly. It is the smooth continuous structural constituent surrounding the branches of the dendrites.


Author(s):  
David A. Ansley

The coherence of the electron flux of a transmission electron microscope (TEM) limits the direct application of deconvolution techniques which have been used successfully on unmanned spacecraft programs. The theory assumes noncoherent illumination. Deconvolution of a TEM micrograph will, therefore, in general produce spurious detail rather than improved resolution.A primary goal of our research is to study the performance of several types of linear spatial filters as a function of specimen contrast, phase, and coherence. We have, therefore, developed a one-dimensional analysis and plotting program to simulate a wide 'range of operating conditions of the TEM, including adjustment of the:(1) Specimen amplitude, phase, and separation(2) Illumination wavelength, half-angle, and tilt(3) Objective lens focal length and aperture width(4) Spherical aberration, defocus, and chromatic aberration focus shift(5) Detector gamma, additive, and multiplicative noise constants(6) Type of spatial filter: linear cosine, linear sine, or deterministic


Author(s):  
J W Steeds

There is a wide range of experimental results related to dislocations in diamond, group IV, II-VI, III-V semiconducting compounds, but few of these come from isolated, well-characterized individual dislocations. We are here concerned with only those results obtained in a transmission electron microscope so that the dislocations responsible were individually imaged. The luminescence properties of the dislocations were studied by cathodoluminescence performed at low temperatures (~30K) achieved by liquid helium cooling. Both spectra and monochromatic cathodoluminescence images have been obtained, in some cases as a function of temperature.There are two aspects of this work. One is mainly of technological significance. By understanding the luminescence properties of dislocations in epitaxial structures, future non-destructive evaluation will be enhanced. The second aim is to arrive at a good detailed understanding of the basic physics associated with carrier recombination near dislocations as revealed by local luminescence properties.


Author(s):  
J.L. Batstone

The development of growth techniques such as metal organic chemical vapor deposition (MOCVD) and molecular beam epitaxy during the last fifteen years has resulted in the growth of high quality epitaxial semiconductor thin films for the semiconductor device industry. The III-V and II-VI semiconductors exhibit a wide range of fundamental band gap energies, enabling the fabrication of sophisticated optoelectronic devices such as lasers and electroluminescent displays. However, the radiative efficiency of such devices is strongly affected by the presence of optically and electrically active defects within the epitaxial layer; thus an understanding of factors influencing the defect densities is required.Extended defects such as dislocations, twins, stacking faults and grain boundaries can occur during epitaxial growth to relieve the misfit strain that builds up. Such defects can nucleate either at surfaces or thin film/substrate interfaces and the growth and nucleation events can be determined by in situ transmission electron microscopy (TEM).


2017 ◽  
Vol 15 (1) ◽  
pp. 21
Author(s):  
Haryo Suganda ◽  
Raja Muhammad Amin

This study is motivated the identification of policies issued by the regional Governmentof Rokan Hulu in the form of Regulatory region number 1 by 2015 on the determination of thevillage and Indigenous Village. Political dynamics based on various interests against themanufacture of, and decision-making in the process of formation of the corresponding localregulations determination of Indigenous Villages in the Rokan Hulu is impacted to a verysignificantamount of changes from the initial draft of the number i.e. 21 (twenty one) the villagebecame Customary 89 (eighty-nine) the Indigenous Villages who have passed. Type of thisresearch is a qualitative descriptive data analysis techniques. The research aims to describe theState of the real situation in a systematic and accurate fact analysis unit or related research, aswell as observations of the field based on the data (information). Method of data collectionwas done with interviews, documentation, and observations through fieldwork (field research).The results of the research on the process of discussion of the draft local regulations andmutual agreement about Designation of Indigenous Villages in the Rokan Hulu is, showed thatthe political dynamics that occur due to the presence of various political interests, rejectionorally by Villagers who were judged to have met the requirements of Draft Regulations to beformulated and the area for the set to be Indigenous Villages, and also there is a desire fromsome villages in the yet to Draft local regulations in order to set the Indigenous village , there isa wide range of interests of these aspects influenced the agreement to assign the entire localVillage which is in the Rokan Hulu become Indigenous village, and the village of Transmigrationinto administrative Villages where the initiator of the changes in the number of IndigenousVillages in the Rokan Hulu it is the desire of the local Government of its own.


1997 ◽  
Vol 3 (S2) ◽  
pp. 81-82
Author(s):  
M.P. Goheen ◽  
M.S. Bartlett ◽  
M.M. Shaw ◽  
S.R. Meshnick ◽  
J.W. Smith

Pneumocystis carinii pneumonia (PCP) occurs at some time in most patients with acquired immunodeficiency syndrome (AIDS). Trimethoprim/sulfamethoxazole or pentamidine isothionate are the traditional modes of therapy for treatment and prophylaxis of PCP. Unfortunately these drugs are associated with a significant incidence of adverse side effects particularly in patients with AIDS. Toxicity and a growing concern that P. carinii strains are becoming resistant to these compounds is providing the impetus for the search for additional drugs to combat P. carinii. Atovaquone, developed as an antimalarial agent, has activity against a wide range of other organisms, including Toxoplasma sp. and P. carinii, with a lower incidence of adverse reactions during clinical trials. Atovaquone inhibits mitochondrial respiration in P. falciparum and P. carinii. In this study transmission electron microscopy (TEM) was used to observe the effects of atovaquone on P. carinii organisms in short term spinner flask culture.Spinner flask cultures of human embryonic lung cells were inoculated with P. carinii from infected rat lung.


2001 ◽  
Vol 19 (4) ◽  
pp. 579-595 ◽  
Author(s):  
D. MOSHER ◽  
B.V. WEBER ◽  
B. MOOSMAN ◽  
R.J. COMMISSO ◽  
P. COLEMAN ◽  
...  

High-sensitivity interferometry measurements of initial density distributions are reviewed for a wide range of gas-puff nozzles used in plasma radiation source (PRS) z-pinch experiments. Accurate gas distributions are required for determining experimental load parameters, modeling implosion dynamics, understanding the radiation properties of the stagnated pinch, and for predicting PRS performance in future experiments. For a number of these nozzles, a simple ballistic-gas-flow model (BFM) has been used to provide good physics-based analytic fits to the measured r, z density distributions. These BFM fits provide a convenient means to smoothly interpolate radial density distributions between discrete axial measurement locations for finer-zoned two-dimensional MHD calculations, and can be used to determine how changes in nozzle parameters and load geometry might alter implosion dynamics and radiation performance. These measurement and analysis techniques are demonstrated for a nested-shell nozzle used in Double Eagle and Saturn experiments. For this nozzle, the analysis suggests load modifications that may increase the K-shell yield.


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