CodedRecon: Video reconstruction for coded exposure imaging techniques

Within the past decade it has become apparent that HVEM offers the biologist a means to explore the three-dimensional structure of cells and/or organelles. Stereo-imaging of thick sections (e.g. 0.25-10 μm) not only reveals anatomical features of cellular components, but also reduces errors of interpretation associated with overlap of structures seen in thick sections. Concomitant with stereo-imaging techniques conventional serial Sectioning methods developed with thin sections have been adopted to serial thick sections (≥ 0.25 μm). Three-dimensional reconstructions of the chondriome of several species of trypanosomatid flagellates have been made from tracings of mitochondrial profiles on cellulose acetate sheets. The sheets are flooded with acetone, gluing them together, and the model sawed from the composite and redrawn.The extensive mitochondrial reticulum can be seen in consecutive thick sections of (0.25 μm thick) Crithidia fasciculata (Figs. 1-2). Profiles of the mitochondrion are distinguishable from the anterior apex of the cell (small arrow, Fig. 1) to the posterior pole (small arrow, Fig. 2).

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Application of Lattice Imaging Techniques to Amorphous Films

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100113664 ◽

1975 ◽

Vol 33 ◽

pp. 8-9

Author(s):

S. R. Herd ◽

P. Chaudhari

Keyword(s):

Nearest Neighbor ◽

Random Network ◽

Imaging Techniques ◽

Network Models ◽

Accurate Method ◽

Direct Transmission ◽

Lattice Imaging ◽

Transmission Electron ◽

Lattice Planes ◽

Nearest Neighbor Distances

Electron diffraction and direct transmission have been used extensively to study the local atomic arrangement in amorphous solids and in particular Ge. Nearest neighbor distances had been calculated from E.D. profiles and the results have been interpreted in terms of the microcrystalline or the random network models. Direct transmission electron microscopy appears the most direct and accurate method to resolve this issue since the spacial resolution of the better instruments are of the order of 3Å. In particular the tilted beam interference method is used regularly to show fringes corresponding to 1.5 to 3Å lattice planes in crystals as resolution tests.

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Studies of metaphase chromosomes in the scanning transmission x-ray microscope: Image fidelity, radiation damage and specimen preparation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100129826 ◽

1992 ◽

Vol 50 (2) ◽

pp. 1038-1039

Author(s):

Shawn Williams ◽

Xiaodong Zhang ◽

Susan Lamm ◽

Jack Van’t Hof

Keyword(s):

Visible Light ◽

Radiation Damage ◽

Cross Section ◽

Imaging Techniques ◽

Dose Range ◽

Specimen Preparation ◽

X Rays ◽

Metaphase Chromosomes ◽

X Ray ◽

Scanning Transmission

The Scanning Transmission X-ray Microscope (STXM) is well suited for investigating metaphase chromosome structure. The absorption cross-section of soft x-rays having energies between the carbon and oxygen K edges (284 - 531 eV) is 6 - 9.5 times greater for organic specimens than for water, which permits one to examine unstained, wet biological specimens with resolution superior to that attainable using visible light. The attenuation length of the x-rays is suitable for imaging micron thick specimens without sectioning. This large difference in cross-section yields good specimen contrast, so that fewer soft x-rays than electrons are required to image wet biological specimens at a given resolution. But most imaging techniques delivering better resolution than visible light produce radiation damage. Soft x-rays are known to be very effective in damaging biological specimens. The STXM is constructed to minimize specimen dose, but it is important to measure the actual damage induced as a function of dose in order to determine the dose range within which radiation damage does not compromise image quality.

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An x-ray microprobe based upon a close-coupled focal-spot / glass capillary arrangement

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100133424 ◽

1992 ◽

Vol 50 (2) ◽

pp. 1758-1759

Author(s):

D. A. Carpenter ◽

M. A. Taylor

Keyword(s):

Imaging Techniques ◽

Fluorescence Analysis ◽

High Sensitivity ◽

Specimen Preparation ◽

X Rays ◽

Glass Capillary ◽

Close Coupling ◽

X Ray ◽

Point To Point ◽

Beam Damage

The development of intense sources of x rays has led to renewed interest in the use of microbeams of x rays in x-ray fluorescence analysis. Sparks pointed out that the use of x rays as a probe offered the advantages of high sensitivity, low detection limits, low beam damage, and large penetration depths with minimal specimen preparation or perturbation. In addition, the option of air operation provided special advantages for examination of hydrated systems or for nondestructive microanalysis of large specimens.The disadvantages of synchrotron sources prompted the development of laboratory-based instrumentation with various schemes to maximize the beam flux while maintaining small point-to-point resolution. Nichols and Ryon developed a microprobe using a rotating anode source and a modified microdiffractometer. Cross and Wherry showed that by close-coupling the x-ray source, specimen, and detector, good intensities could be obtained for beam sizes between 30 and 100μm. More importantly, both groups combined specimen scanning with modern imaging techniques for rapid element mapping.

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SEM/FESEM imaging of lamellar structures in melt extruded polyethylene films

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100130341 ◽

1992 ◽

Vol 50 (2) ◽

pp. 1142-1143

Author(s):

R.T. Chen ◽

M.G. Jamieson ◽

R. Callahan

Keyword(s):

Imaging Techniques ◽

Membrane Surface ◽

High Stress ◽

Extrusion Direction ◽

Polymeric Membranes ◽

Scanning Tunneling ◽

Tunneling Microscopy ◽

Crystalline Polymers ◽

Lamellar Structures ◽

Resolution Imaging

“Row lamellar” structures have previously been observed when highly crystalline polymers are melt-extruded and recrystallized under high stress. With annealing to perfect the stacked lamellar superstructure and subsequent stretching in the machine (extrusion) direction, slit-like micropores form between the stacked lamellae. This process has been adopted to produce polymeric membranes on a commercial scale with controlled microporous structures. In order to produce the desired pore morphology, row lamellar structures must be established in the membrane precursors, i.e., as-extruded and annealed polymer films or hollow fibers. Due to the lack of pronounced surface topography, the lamellar structures have typically been investigated by replica-TEM, an indirect and time consuming procedure. Recently, with the availability of high resolution imaging techniques such as scanning tunneling microscopy (STM) and field emission scanning electron microscopy (FESEM), the microporous structures on the membrane surface as well as lamellar structures in the precursors can be directly examined.The materials investigated are Celgard® polyethylene (PE) flat sheet membranes and their film precursors, both as-extruded and annealed, made at different extrusion rates (E.R.).

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High-resolution topographic SEM and radiation damage: The rationale for using low beam voltage

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100131024 ◽

1992 ◽

Vol 50 (2) ◽

pp. 1278-1279

Author(s):

James Pawley ◽

David Joy

Keyword(s):

High Resolution ◽

Imaging Techniques ◽

Morphological Structure ◽

Radiation Sensitivity ◽

Beam Specimen ◽

Measured Signal ◽

Practical Consideration ◽

Interaction Volume ◽

Impact Area ◽

Signal Contrast

The scanning electron microscope (SEM) builds up an image by sampling contiguous sub-volumes near the surface of the specimen. A fine electron beam selectively excites each sub-volume and then the intensity of some resulting signal is measured and then plotted as a corresponding intensity in an image. The spatial resolution of such an image is limited by at least three factors. Two of these determine the size of the interaction volume: the size of the electron probe and the extent to which detectable signal is excited from locations remote from the beam impact area. A third limitation emerges from the fact that the probing beam is composed of a number of discrete particles and therefore that the accuracy with which any detectable signal can be measured is limited by Poisson statistics applied to this number (or to the number of events actually detected if this is smaller). As in all imaging techniques, the limiting signal contrast required to recognize a morphological structure is constrained by this statistical consideration. The only way to overcome this limit is to increase either the contrast of the measured signal or the number of beam/specimen interactions detected. Unfortunately, these interactions deposit ionizing radiation that may damage the very structure under investigation. As a result, any practical consideration of the high resolution performance of the SEM must consider not only the size of the interaction volume but also the contrast available from the signal producing the image and the radiation sensitivity of the specimen.

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Quantitative nano-characterization of heterogeneous catalysts

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100138361 ◽

1995 ◽

Vol 53 ◽

pp. 398-399

Author(s):

P.A. Crozier ◽

M. Pan

Keyword(s):

Heterogeneous Catalysts ◽

Low Dose ◽

Imaging Techniques ◽

Structural Features ◽

Catalyst Characterization ◽

New Developments ◽

Double Phase ◽

Phase Systems ◽

Dose Imaging

Heterogeneous catalysts can be of varying complexity ranging from single or double phase systems to complicated mixtures of metals and oxides with additives to help promote chemical reactions, extend the life of the catalysts, prevent poisoning etc. Although catalysis occurs on the surface of most systems, detailed descriptions of the microstructure and chemistry of catalysts can be helpful for developing an understanding of the mechanism by which a catalyst facilitates a reaction. Recent years have seen continued development and improvement of various TEM, STEM and AEM techniques for yielding information on the structure and chemistry of catalysts on the nanometer scale. Here we review some quantitative approaches to catalyst characterization that have resulted from new developments in instrumentation.HREM has been used to examine structural features of catalysts often by employing profile imaging techniques to study atomic details on the surface. Digital recording techniques employing slow-scan CCD cameras have facilitated the use of low-dose imaging in zeolite structure analysis and electron crystallography. Fig. la shows a low-dose image from SSZ-33 zeolite revealing the presence of a stacking fault.

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Growth of hematite on (0001) and {1102} sapphire substrates

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100175016 ◽

1990 ◽

Vol 48 (4) ◽

pp. 376-377

Author(s):

Lisa A. Tietz ◽

Scott R. Summerfelt ◽

C. Barry Carter

Keyword(s):

Imaging Techniques ◽

Chemical Vapor ◽

Interface Energy ◽

Dark Field ◽

Cation Sublattice ◽

Sapphire Substrates ◽

Dark Field Imaging ◽

Weak Beam ◽

Pressure Chemical ◽

Interface Structures

Defects in thin films are often introduced at the substrate-film interface during the early stages of growth. The interface structures of semiconductor heterojunctions have been extensively studied because of the electrical activity of defects in these materials. Much less attention has been paid to the structure of oxide-oxide heterojunctions. In this study, the structures of the interfaces formed between hematite (α-Fe2O3) and two orientations of sapphire (α-Al2O3) are examined in relationship to the defects introduced into the hematite film. In such heterojunctions, the oxygen sublattice is expected to have a strong influence on the epitaxy; however, defects which involve only the cation sublattice may be introduced at the interface with little increase in interface energy.Oxide heterojunctions were produced by depositing small quantities of hematite directly onto electrontransparent sapphire substrates using low-pressure chemical vapor deposition. Prior to deposition, the ionthinned substrates were chemically cleaned and annealed at 1400°C to give “clean”, crystalline surfaces. Hematite was formed by the reaction of FeCl3 vapor with water vapor at 1150°C and 1-2 Torr. The growth of the hematite and the interface structures formed on (0001) and {102} substrates have been studied by bright-field, strong- and weak-beam dark-field imaging techniques.

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Recent applications of TEM to the study of interfaces

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100174679 ◽

1990 ◽

Vol 48 (4) ◽

pp. 308-309

Author(s):

C. Barry Carter

Keyword(s):

High Resolution ◽

Sample Preparation ◽

Grain Boundaries ◽

Diffuse Scattering ◽

Imaging Techniques ◽

Amorphous Material ◽

Contrast Imaging ◽

Current State ◽

Actual Nature ◽

High Resolution Images

This paper will review the current state of understanding of interface structure and highlight some of the future needs and problems which must be overcome. The study of this subject can be separated into three different topics: 1) the fundamental electron microscopy aspects, 2) material-specific features of the study and 3) the characteristics of the particular interfaces. The two topics which are relevant to most studies are the choice of imaging techniques and sample preparation. The techniques used to study interfaces in the TEM include high-resolution imaging, conventional diffraction-contrast imaging, and phase-contrast imaging (Fresnel fringe images, diffuse scattering). The material studied affects not only the characteristics of the interfaces (through changes in bonding, etc.) but also the method used for sample preparation which may in turn have a significant affect on the resulting image. Finally, the actual nature and geometry of the interface must be considered. For example, it has become increasingly clear that the plane of the interface is particularly important whenever at least one of the adjoining grains is crystalline.A particularly productive approach to the study of interfaces is to combine different imaging techniques as illustrated in the study of grain boundaries in alumina. In this case, the conventional imaging approach showed that most grain boundaries in ion-thinned samples are grooved at the grain boundary although the extent of this grooving clearly depends on the crystallography of the surface. The use of diffuse scattering (from amorphous regions) gives invaluable information here since it can be used to confirm directly that surface grooving does occur and that the grooves can fill with amorphous material during sample preparation (see Fig. 1). Extensive use of image simulation has shown that, although information concerning the interface can be obtained from Fresnel-fringe images, the introduction of artifacts through sample preparation cannot be lightly ignored. The Fresnel-fringe simulation has been carried out using a commercial multislice program (TEMPAS) which was intended for simulation of high-resolution images.

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