Applications of Through-Focus Dark Field Image Shifts to Phase Identification

1975 ◽  
Vol 33 ◽  
pp. 178-179
Author(s):  
Prakash Rao
Keyword(s):  
Dark Field Image ◽  
Thin Foil ◽  
Dark Field ◽  
The Other ◽  
Stereo Image ◽  
Phase Identification ◽  
The Past ◽  
Field Electron Microscopy ◽  
Dark Field Electron ◽  
Recent Extension

Image shifts in out-of-focus dark field images have been used in the past to determine, for example, epitaxial relationships in thin films. A recent extension of the use of dark field image shifts has been to out-of-focus images in conjunction with stereoviewing to produce an artificial stereo image effect. The technique, called through-focus dark field electron microscopy or 2-1/2D microscopy, basically involves obtaining two beam-tilted dark field images such that one is slightly over-focus and the other slightly under-focus, followed by examination of the two images through a conventional stereoviewer. The elevation differences so produced are usually unrelated to object positions in the thin foil and no specimen tilting is required.In order to produce this artificial stereo effect for the purpose of phase separation and identification, it is first necessary to select a region of the diffraction pattern containing more than just one discrete spot, with the objective aperture.

Imaging a Protein α-Helix by Dark Field Electron Microscopy

1981 ◽  
Vol 39 ◽  
pp. 34-35
Author(s):  
D. W. Andrews ◽  
F. P. Ottensmeyer
Keyword(s):  
Electron Microscopy ◽  
Visual Recognition ◽  
Dark Field Image ◽  
Relevant Information ◽  
Dark Field ◽  
Biologically Relevant ◽  
Field Electron ◽  
Field Electron Microscopy ◽  
Dark Field Electron ◽  
Α Helix

Elucidation of the fine structure of a protein, beyond its gross conformation, requires images of better than 10 Å resolution. In order to achieve such a resolution the normal techniques of staining the molecule must be avoided by substituting a technique such as dark field electron microscopy. However, to form a dark field image of such high resolution requires an exposure of approximately 1500 e/Å2. Exposures of this magnitude are so high that serious doubt has been expressed regarding the extent to which an image formed under these circumstances is an accurate representation of the molecule. Images of myokinase, protamine, glucagon, secretin, and ACTH indicate that reproducible 5-10 Å detail is present in the micrographs. Furthermore, for bacitrin, valinomycin, and the growth stimulators LMW-CSA N and B, biologically relevant information of 3-5 Å is present in averages prepared from a collection of images or even in single molecular images. Visual recognition has been used as a criterion to assess radiation damage in images of isolated molecules of protamine.

Dynamic Dark-Field Electron Microscopy

1976 ◽  
Vol 34 ◽  
pp. 482-483
Author(s):  
G.B. Haydon ◽  
R.A. Crane ◽  
C.R. Zercher
Keyword(s):  
Electron Microscopy ◽  
Optical Axis ◽  
Dark Field Image ◽  
Dark Field ◽  
Annular Ring ◽  
Field Electron ◽  
Field Electron Microscopy ◽  
Dark Field Electron

Dynamic dark-field electron microscopy as described here provides capabilities not present with other methods. An annular ring of any selected diameter is used to illuminate the specimen. Diffraction rings selected by the objective aperture are integrated photographically to produce the dark-field image.All methods of dark-field electron microscopy eliminate the incident illumination from the image and utilize only a selected portion of the scattered electrons and each has its limitations (1):1. Movement of the objective aperture off of the optical axis introduces spherical aberation which increases as the 4th power of the distance from the axis.2. Tilting the beam either mechanically or electrically allows only those electrons scattered in one direction to be imaged.

Utilizing Dark Field Electron Microscopy to Produce Lantern Slides

1974 ◽  
Vol 32 ◽  
pp. 116-117
Author(s):  
J. N. Meador ◽  
C. N. Sun ◽  
H. J. White
Keyword(s):  
Electron Microscope ◽  
Electron Micrographs ◽  
Dark Field ◽  
Limiting Factor ◽  
Clinical Laboratories ◽  
Field Electron ◽  
Time Element ◽  
Field Electron Microscopy ◽  
Dark Field Electron ◽  
Dark Field Micrographs

The electron microscope is being utilized more and more in clinical laboratories for pathologic diagnosis. One of the major problems in the utilization of the electron microscope for diagnostic purposes is the time element involved. Recent experimentation with rapid embedding has shown that this long phase of the process can be greatly shortened. In rush cases the making of projection slides can be eliminated by taking dark field electron micrographs which show up as a positive ready for use. The major limiting factor for use of dark field micrographs is resolution. However, for conference purposes electron micrographs are usually taken at 2.500X to 8.000X. At these low magnifications the resolution obtained is quite acceptable.

Protamine-DNA interaction

1978 ◽  
Vol 36 (2) ◽  
pp. 200-201
Author(s):  
D.P. Bazett-Jones ◽  
F.P. Ottensmeyer
Keyword(s):  
Small Volume ◽  
Double Helix ◽  
Dna Interaction ◽  
Dark Field ◽  
Sperm Head ◽  
Nuclear Proteins ◽  
Field Electron Microscopy ◽  
Dark Field Electron ◽  
Dna Double Helix ◽  
Positively Charged

Dark field electron microscopy has been used for the study of the structure of individual macromolecules with a resolution to at least the 5Å level. The use of this technique has been extended to the investigation of structure of interacting molecules, particularly the interaction between DNA and fish protamine, a class of basic nuclear proteins of molecular weight 4,000 daltons.Protamine, which is synthesized during spermatogenesis, binds to chromatin, displaces the somatic histones and wraps up the DNA to fit into the small volume of the sperm head. It has been proposed that protamine, existing as an extended polypeptide, winds around the minor groove of the DNA double helix, with protamine's positively-charged arginines lining up with the negatively-charged phosphates of DNA. However, viewing protamine as an extended protein is inconsistent with the results obtained in our laboratory.

Evaluation of the dark field method for large association of spread DNA

1978 ◽  
Vol 36 (2) ◽  
pp. 190-191
Author(s):  
Douglas C. Barker
Keyword(s):  
Electron Microscopy ◽  
Molecular Weight ◽  
Mitochondrial Dna ◽  
Extraction Method ◽  
Field Method ◽  
Dark Field ◽  
Kinetoplast Dna ◽  
Field Electron ◽  
Field Electron Microscopy ◽  
Dark Field Electron

A number of satisfactory methods are available for the electron microscopy of nicleic acids. These methods concentrated on fragments of nuclear, viral and mitochondrial DNA less than 50 megadaltons, on denaturation and heteroduplex mapping (Davies et al 1971) or on the interaction between proteins and DNA (Brack and Delain 1975). Less attention has been paid to the experimental criteria necessary for spreading and visualisation by dark field electron microscopy of large intact issociations of DNA. This communication will report on those criteria in relation to the ultrastructure of the (approx. 1 x 10-14g) DNA component of the kinetoplast from Trypanosomes. An extraction method has been developed to eliminate native endonucleases and nuclear contamination and to isolate the kinetoplast DNA (KDNA) as a compact network of high molecular weight. In collaboration with Dr. Ch. Brack (Basel [nstitute of Immunology), we studied the conditions necessary to prepare this KDNA Tor dark field electron microscopy using the microdrop spreading technique.

Thin Pyrolytic Graphite Films for Electron Microscope Substrates

1971 ◽  
Vol 29 ◽  
pp. 226-227 ◽  
Author(s):  
George H. N. Riddle ◽  
Benjamin M. Siegel
Keyword(s):  
Vacuum Chamber ◽  
Pyrolytic Graphite ◽  
High Vacuum ◽  
Dark Field ◽  
Ultra High Vacuum ◽  
Graphite Substrate ◽  
Nickel Substrate ◽  
Routine Procedure ◽  
Field Electron Microscopy ◽  
Dark Field Electron

A routine procedure for growing very thin graphite substrate films has been developed. The films are grown pyrolytically in an ultra-high vacuum chamber by exposing (111) epitaxial nickel films to carbon monoxide gas. The nickel serves as a catalyst for the disproportionation of CO through the reaction 2C0 → C + CO2. The nickel catalyst is prepared by evaporation onto artificial mica at 400°C and annealing for 1/2 hour at 600°C in vacuum. Exposure of the annealed nickel to 1 torr CO for 3 hours at 500°C results in the growth of very thin continuous graphite films. The graphite is stripped from its nickel substrate in acid and mounted on holey formvar support films for use as specimen substrates.The graphite films, self-supporting over formvar holes up to five microns in diameter, have been studied by bright and dark field electron microscopy, by electron diffraction, and have been shadowed to reveal their topography and thickness. The films consist of individual crystallites typically a micron across with their basal planes parallel to the surface but oriented in different, apparently random directions about the normal to the basal plane.

Characteristic Geometry and Diffraction Contrast of Dispersed ThO2 Crystals

1969 ◽  
Vol 27 ◽  
pp. 168-169
Author(s):  
R. J. Horylev ◽  
L. E. Murr
Keyword(s):  
Dark Field ◽  
Particle Orientation ◽  
Contrast Effects ◽  
Strain Fields ◽  
Diffraction Contrast ◽  
Dispersed Particles ◽  
Field Electron Microscopy ◽  
Dark Field Electron ◽  
Particle Images ◽  
Diffraction Effects

Smith has shown by dark-field electron microscopy of extracted ThO2 particles from TD-nickel (2% ThO2) that they possess single crystal characteristics. It is generally assumed that these particle dispersions are incoherent. However, some diffraction effects associated with the particle images appeared to be similar to coherency strain fields. The present work will demonstrate conclusively that ThO2 dispersed particles in TD-nickel (2% ThO2) and TD-NiCr (2% ThO2, 20% Cr, Ni) are single crystals. Moreover, the diffraction contrast effects are extinction fringes. That is, these effects arise because of the particle orientation with respect to the electron beam and the extinction conditions for various operating reflections The particles are in fact incoherent.

Effets of surfaces on precipitate morphology in irradiated thin foils

1978 ◽  
Vol 36 (1) ◽  
pp. 654-655
Author(s):  
D.I. Potter ◽  
A. Taylor
Keyword(s):  
Ion Irradiation ◽  
Dark Field Image ◽  
Thin Foil ◽  
Dark Field ◽  
Al Alloy ◽  
Bright Field Image ◽  
Dislocation Loops ◽  
Precipitate Morphology ◽  
Thin Foils

Thermal aging of Ni-12.8 at. % A1 and Ni-12.7 at. % Si produces spatially homogeneous dispersions of cuboidal γ'-Ni3Al or Ni3Si precipitate particles arrayed in the Ni solid solution. We have used 3.5-MeV 58Ni+ ion irradiation to examine the effect of irradiation during precipitation on precipitate morphology and distribution. The nearness of free surfaces produced unusual morphologies in foils thinned prior to irradiation. These thin-foil effects will be important during in-situ investigations of precipitation in the HVEM. The thin foil results can be interpreted in terms of observations from bulk irradiations which are described first.Figure 1a is a dark field image of the γ' precipitate 5000 Å beneath the surface(∿1200 Å short of peak damage) of the Ni-Al alloy irradiated in bulk form. The inhomogeneous spatial distribution of γ' results from the presence of voids and dislocation loops which can be seen in the bright field image of the same area, Fig. 1b.

Structure and Interaction of Protamine by Dark Field Electron Microscopy

1976 ◽  
Vol 34 ◽  
pp. 160-161
Author(s):  
D.P. Bazett-Jones ◽  
F.P. Ottensmeyer
Keyword(s):  
Electron Microscopy ◽  
Dark Field ◽  
Amino Acid Sequences ◽  
Dimensional Structure ◽  
3 Dimensional ◽  
Field Electron ◽  
Proposed Model ◽  
Field Electron Microscopy ◽  
Dark Field Electron ◽  
Positively Charged

It has been shown for some time that it is possible to obtain images of small unstained proteins, with a resolution of approximately 5Å using dark field electron microscopy (1,2). Applying this technique, we have observed a uniformity in size and shape of the 2-dimensional images of pure specimens of fish protamines (salmon, herring (clupeine, Y-l) and rainbow trout (Salmo irideus)). On the basis of these images, a model for the 3-dimensional structure of the fish protamines has been proposed (2).The known amino acid sequences of fish protamines show stretches of positively charged arginines, separated by regions of neutral amino acids (3). The proposed model for protamine structure (2) consists of an irregular, right-handed helix with the segments of adjacent arginines forming the loops of the coil.

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