An Image Reconstruction System Applied to High Voltage Microscopy

1975 ◽  
Vol 33 ◽  
pp. 292-293
Author(s):  
D. E. Johnson
Keyword(s):  
High Voltage ◽  
Prior Information ◽  
Block Diagram ◽  
Three Dimensional ◽  
Real Space ◽  
Two Dimensional ◽  
Efficient Manner ◽  
Fourier Methods ◽  
Tilt Series ◽  
Methods Of Reconstruction

Increased specimen penetration; the principle advantage of high voltage microscopy, is accompanied by an increased need to utilize information on three dimensional specimen structure available in the form of two dimensional projections (i.e. micrographs). We are engaged in a program to develop methods which allow the maximum use of information contained in a through tilt series of micrographs to determine three dimensional speciman structure.In general, we are dealing with structures lacking in symmetry and with projections available from only a limited span of angles (±60°). For these reasons, we must make maximum use of any prior information available about the specimen. To do this in the most efficient manner, we have concentrated on iterative, real space methods rather than Fourier methods of reconstruction. The particular iterative algorithm we have developed is given in detail in ref. 3. A block diagram of the complete reconstruction system is shown in fig. 1.

Three-Dimensional Reconstruction of Two Forms of the Chaperonin GRO EL

1990 ◽  
Vol 48 (1) ◽  
pp. 258-259
Author(s):  
J.L. Carrascosa ◽  
G. Abella ◽  
S. Marco ◽  
M. Muyal ◽  
J.M. Carazo
Keyword(s):  
Three Dimensional ◽  
The Other ◽  
Two Dimensional ◽  
Series Method ◽  
Three Dimensional Reconstruction ◽  
Structural Components ◽  
E Coli ◽  
Dimensional Reconstruction ◽  
Morphogenetic Factors ◽  
Tilt Series

Chaperonins are a class of proteins characterized by their role as morphogenetic factors. They trantsiently interact with the structural components of certain biological aggregates (viruses, enzymes etc), promoting their correct folding, assembly and, eventually transport. The groEL factor from E. coli is a conspicuous member of the chaperonins, as it promotes the assembly and morphogenesis of bacterial oligomers and/viral structures.We have studied groEL-like factors from two different bacteria:E. coli and B.subtilis. These factors share common morphological features , showing two different views: one is 6-fold, while the other shows 7 morphological units. There is also a correlation between the presence of a dominant 6-fold view and the fact of both bacteria been grown at low temperature (32°C), while the 7-fold is the main view at higher temperatures (42°C). As the two-dimensional projections of groEL were difficult to interprete, we studied their three-dimensional reconstruction by the random conical tilt series method from negatively stained particles.

scholarly journals 6D electron microscopy: combining real-space and reciprocal-space tomography

2014 ◽  
Vol 70 (a1) ◽  
pp. C368-C368 ◽  
Author(s):  
Alexander Eggeman ◽  
Robert Krakow ◽  
Paul Midgley
Keyword(s):  
Reciprocal Lattice ◽  
Three Dimensional ◽  
Real Space ◽  
Dark Field ◽  
Reciprocal Space ◽  
Data Set ◽  
Wide Range ◽  
The Matrix ◽  
Precession Electron Diffraction ◽  
Tilt Series

STEM and TEM-based tomography has been used widely to study the 3D morphology of a wide range of materials. Similarly reciprocal space tomography in which a tilt-series of diffraction patterns are acquired offers a powerful method for the analysis of the atomic structure of crystalline materials. The natural progression is to combine these techniques into a complete three dimensional morphology and crystallography data set, allowing both features to be studied simultaneously. Using a tilt series of scanning precession electron diffraction measurements from a commercially available Ni-base superalloy as an example, the complete reciprocal lattice orientation for a number of components embedded within the matrix could be determined. It was straightforward to identify reciprocal lattice vectors that allowed dark-field images representing each phase to be produced post-acquisition. In turn these were combined using geometric tomography methods to yield a 3-D tomogram of the superalloy. Imaging these phases using conventional ADF STEM tomography would potentially be challenging given the compositional similarity between the different phases. From the combined dataset the spatial distribution of the component phases could be easily recovered but more importantly the orientational relationships between these different components could be unambiguously determined. In this way the thermo-mechanical history of the sample could be inferred from the arrangement of coherent and semi-coherent interfaces and a previously unreported crystallographic registry between metal carbide (MC) and the matrix f.c.c. phases could been identified. The possibilities for development and applications of this technique will be discussed further.

Imaging of strain and lattice orientation by quick scanning X-ray microscopy combined with three-dimensional reciprocal space mapping

2014 ◽  
Vol 47 (2) ◽  
pp. 762-769 ◽  
Author(s):  
Gilbert André Chahine ◽  
Marie-Ingrid Richard ◽  
Roberto Arturo Homs-Regojo ◽  
Thu Nhi Tran-Caliste ◽  
Dina Carbone ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Continuous Mapping ◽  
Real Space ◽  
Reciprocal Space ◽  
Data Sets ◽  
Two Dimensional ◽  
Reciprocal Space Mapping ◽  
X Ray ◽  
Study Materials ◽  
User Friendly

Numerous imaging methods have been developed over recent years in order to study materials at the nanoscale. Within this context, scanning X-ray diffraction microscopy has become a routine technique, giving access to structural properties with sub-micrometre resolution. This article presents an optimized technique and an associated software package which have been implemented at the ID01 beamline (ESRF, Grenoble). A structural scanning probe microscope with intriguing imaging qualities is obtained. The technique consists in a two-dimensional quick continuous mapping with sub-micrometre resolution of a sample at a given reciprocal space position. These real space maps are made by continuously moving the sample while recording scattering images with a fast two-dimensional detector for every point along a rocking curve. Five-dimensional data sets are then produced, consisting of millions of detector images. The images are processed by the user-friendly X-ray strain orientation calculation software (XSOCS), which has been developed at ID01 for automatic analysis. It separates tilt and strain and generates two-dimensional maps of these parameters. At spatial resolutions of typically 200–800 nm, this quick imaging technique achieves strain sensitivity below Δa/a= 10−5and a resolution of tilt variations down to 10−3° over a field of view of 100 × 100 µm.

Image Reconstruction Applied to High Voltage Microscopy

1974 ◽  
Vol 32 ◽  
pp. 396-397
Author(s):  
D. E. Johnson ◽  
J. Pfeifer
Keyword(s):  
Electron Microscopy ◽  
Image Reconstruction ◽  
High Voltage ◽  
Three Dimensional ◽  
Two Dimensional ◽  
Three Dimensional Reconstruction ◽  
High Voltage Electron Microscopy ◽  
Voltage Electron ◽  
Dimensional Reconstruction ◽  
Quantitative Results

Greatly increased specimen penetration, which is the principle advantage of high voltage electron microscopy, carries with it an increased need for techniques to interpret the large amount of three-dimensional information projected into two-dimensional micrographs. Stereo views can provide very useful information and are widely used. However, for the general specimen, stereo views are limited in their ability to produce quantitative results. At the high voltage microscope facility, Univ. of Wisconsin, we have begun a program to develop and apply three dimensional reconstruction techniques to the microscopy of 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.

High Resolution Microscopy of Multi-Layered Biological Crystals

1982 ◽  
Vol 40 ◽  
pp. 80-83
Author(s):  
H.A. Cohen ◽  
T.W. Jeng ◽  
W. Chiu
Keyword(s):  
High Resolution ◽  
Low Dose ◽  
Three Dimensional ◽  
Data Interpretation ◽  
Two Dimensional ◽  
Biological Objects ◽  
Density Maps ◽  
Density Map ◽  
Complex Crystal ◽  
High Resolution Microscopy

This tutorial will discuss the methodology of low dose electron diffraction and imaging of crystalline biological objects, the problems of data interpretation for two-dimensional projected density maps of glucose embedded protein crystals, the factors to be considered in combining tilt data from three-dimensional crystals, and finally, the prospects of achieving a high resolution three-dimensional density map of a biological crystal. This methodology will be illustrated using two proteins under investigation in our laboratory, the T4 DNA helix destabilizing protein gp32*I and the crotoxin complex crystal.

Instrumentation For Quantitative Microscopy

1977 ◽  
Vol 35 ◽  
pp. 206-209
Author(s):  
B. Ralph ◽  
A.R. Jones
Keyword(s):  
Volume Fraction ◽  
Three Dimensional ◽  
Two Dimensional ◽  
Automatic Data ◽  
Phase Volume Fraction ◽  
Statistical Confidence ◽  
Resultant Data ◽  
Automatic Data Collection ◽  
Third Dimension ◽  
Evolution Of Microstructure

In all fields of microscopy there is an increasing interest in the quantification of microstructure. This interest may stem from a desire to establish quality control parameters or may have a more fundamental requirement involving the derivation of parameters which partially or completely define the three dimensional nature of the microstructure. This latter categorey of study may arise from an interest in the evolution of microstructure or from a desire to generate detailed property/microstructure relationships. In the more fundamental studies some convolution of two-dimensional data into the third dimension (stereological analysis) will be necessary.In some cases the two-dimensional data may be acquired relatively easily without recourse to automatic data collection and further, it may prove possible to perform the data reduction and analysis relatively easily. In such cases the only recourse to machines may well be in establishing the statistical confidence of the resultant data. Such relatively straightforward studies tend to result from acquiring data on the whole assemblage of features making up the microstructure. In this field data mode, when parameters such as phase volume fraction, mean size etc. are sought, the main case for resorting to automation is in order to perform repetitive analyses since each analysis is relatively easily performed.

A New 1000kv Electron Microscope

1969 ◽  
Vol 27 ◽  
pp. 100-101
Author(s):  
J.L. Williams ◽  
K. Heathcote ◽  
E.J. Greer
Keyword(s):  
Electron Microscope ◽  
Direct Observation ◽  
High Voltage ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Specimen Thickness ◽  
Voltage Electron ◽  
High Voltage Electron Microscope ◽  
Dynamic Experiments ◽  
Conventional Instrument

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.

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.

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