Maximum entropy EM tomography: demonstration of improved resolution with adenovirus

Author(s):  
Ulf Skoglund ◽  
Lars-Göran Öfverstedt ◽  
Roger Burnett ◽  
Gérard Bricogne

EMT is a method for three-dimensional (3-D) reconstruction of single objects from electron microscope pictures in a tilt series. The EMT method is general and can be applied to any transparent object, and is not restricted to symmetrical or regularly arranged objects, nor to objects with preferred orientations on a support grid. In its present shape, the EMT method allows reproducible 3-D reconstructions of molecular objects with a resolution in the range of 5 nm. Currently, the EMT method covers the intermediate resolution range where there is no other physical technique available to analyze single molecular complexes.The recent availability of techniques for high quality automatic data collection from vitrified specimens at low dose promises an improvement of the reproducible resolution as well as considerable expansion of the number of users of the EMT technique. A serious drawback of EMT, however, has been the notoriously low quality of single-object 3-D reconstructions when compared to symmetrized reconstructions.

Author(s):  
B. Ralph ◽  
A.R. Jones

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.


2021 ◽  
Author(s):  
Takashi Morita ◽  
Aru Toyoda ◽  
Seitaro Aisu ◽  
Naoko Suda-Hashimoto ◽  
Akihisa Kaneko ◽  
...  

One of the goals in animal socioecology is to understand the functions and dynamics of group living. While observations of free-ranging animals are a crucial source of information, an experimental investigation that manipulates the size or composition, or both, of animal groups in captivity can also bring complementary contributions to the research inquiry. When paired with an automatic data collection by biologging technology, experimental studies on captive animals also allow for big data analyses based on recent machine learning techniques. As an initial exploration of this research paradigm, the present study inquired to what extent isolation of captive Japanese macaques (Macaca fuscata) changed their movement patterns. Using three-dimensional location trajectories of the macaques that were systematically collected via Bluetooth Low Energy beacons and a deep neural network, we estimated the identifiability of whether a macaque was behaving in isolation or in group. We found that the neural network identified the isolation vs. in-group conditions with more than 90% accuracy from a five-minute location trajectory, suggesting that the isolation caused notable changes from the canonical group-living behaviors. In addition, the isolation made each individual more identifiable from one another based on their location trajectories.


2021 ◽  
Vol 235 ◽  
pp. 03084
Author(s):  
Lei Ma ◽  
Jiachen Che ◽  
Xiao Qian ◽  
Hongfang Song ◽  
Geping Wen ◽  
...  

As a new decentralized infrastructure and distributed computing paradigm, blockchain is of great significance to break through the “mutual trust problem” between statistical specialty and basic data providing departments, and between the upper and lower levels of statistical data reporting units, and further improve the quality of statistical work. This paper first briefly analyzes the problems existing in the current investment statistics business, and introduces the basic principles and characteristics of blockchain technology. This paper studies the idea and process optimization of the application of blockchain technology in investment statistics business, and expounds the specific application schemes of blockchain technology in different links, such as automatic data collection, automatic data verification, automatic report generation, and automatic index release. Finally, the expected effect of the application of blockchain technology in investment statistics business is analyzed.


Author(s):  
H.A. Cohen ◽  
T.W. Jeng ◽  
W. Chiu

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.


Author(s):  
S. Khadpe ◽  
R. Faryniak

The Scanning Electron Microscope (SEM) is an important tool in Thick Film Hybrid Microcircuits Manufacturing because of its large depth of focus and three dimensional capability. This paper discusses some of the important areas in which the SEM is used to monitor process control and component failure modes during the various stages of manufacture of a typical hybrid microcircuit.Figure 1 shows a thick film hybrid microcircuit used in a Motorola Paging Receiver. The circuit consists of thick film resistors and conductors screened and fired on a ceramic (aluminum oxide) substrate. Two integrated circuit dice are bonded to the conductors by means of conductive epoxy and electrical connections from each integrated circuit to the substrate are made by ultrasonically bonding 1 mil aluminum wires from the die pads to appropriate conductor pads on the substrate. In addition to the integrated circuits and the resistors, the circuit includes seven chip capacitors soldered onto the substrate. Some of the important considerations involved in the selection and reliability aspects of the hybrid circuit components are: (a) the quality of the substrate; (b) the surface structure of the thick film conductors; (c) the metallization characteristics of the integrated circuit; and (d) the quality of the wire bond interconnections.


Author(s):  
B. Carragher ◽  
M. Whittaker

Techniques for three-dimensional reconstruction of macromolecular complexes from electron micrographs have been successfully used for many years. These include methods which take advantage of the natural symmetry properties of the structure (for example helical or icosahedral) as well as those that use single axis or other tilting geometries to reconstruct from a set of projection images. These techniques have traditionally relied on a very experienced operator to manually perform the often numerous and time consuming steps required to obtain the final reconstruction. While the guidance and oversight of an experienced and critical operator will always be an essential component of these techniques, recent advances in computer technology, microprocessor controlled microscopes and the availability of high quality CCD cameras have provided the means to automate many of the individual steps.During the acquisition of data automation provides benefits not only in terms of convenience and time saving but also in circumstances where manual procedures limit the quality of the final reconstruction.


Author(s):  
Weiping Liu ◽  
John W. Sedat ◽  
David A. Agard

Any real world object is three-dimensional. The principle of tomography, which reconstructs the 3-D structure of an object from its 2-D projections of different view angles has found application in many disciplines. Electron Microscopic (EM) tomography on non-ordered structures (e.g., subcellular structures in biology and non-crystalline structures in material science) has been exercised sporadically in the last twenty years or so. As vital as is the 3-D structural information and with no existing alternative 3-D imaging technique to compete in its high resolution range, the technique to date remains the kingdom of a brave few. Its tedious tasks have been preventing it from being a routine tool. One keyword in promoting its popularity is automation: The data collection has been automated in our lab, which can routinely yield a data set of over 100 projections in the matter of a few hours. Now the image processing part is also automated. Such automations finish the job easier, faster and better.


Author(s):  
G. Lembcke ◽  
F. Zemlin

The thermoacidophilic archaebacterium Sulfolobus spec. B12 , which is closely related to Sulfolobus solfataricus , possesses a regularly arrayed surface protein (S-layer), which is linked to the plasma membrane via spacer elements spanning a distinct interspace of approximately 18 nm. The S-layer has p3-Symmetry and a lattice constant of 21 nm; three-dimensional reconstructions of negatively stained fragments yield a layer thickness of approximately 6-7 nm.For analysing the molecular architecture of Sulfolobus surface protein in greater detail we use aurothioglucose(ATG)-embedding for specimen preparation. Like glucose, ATG, is supposed to mimic the effect of water, but has the advantage of being less volatile. ATG has advantages over glucose when working with specimens composed exclusively of protein because of its higher density of 2.92 g cm-3. Because of its high radiation sensitivity electromicrographs has to be recorded under strict low-dose conditions. We have recorded electromicrographs with a liquid helium-cooled superconducting electron microscope (the socalled SULEIKA at the Fritz-Haber-lnstitut) with a specimen temperature of 4.5 K and with a maximum dose of 2000 e nm-2 avoiding any pre-irradiation of the specimen.


1990 ◽  
Vol 18 (4) ◽  
pp. 216-235 ◽  
Author(s):  
J. De Eskinazi ◽  
K. Ishihara ◽  
H. Volk ◽  
T. C. Warholic

Abstract The paper describes the intention of the authors to determine whether it is possible to predict relative belt edge endurance for radial passenger car tires using the finite element method. Three groups of tires with different belt edge configurations were tested on a fleet test in an attempt to validate predictions from the finite element results. A two-dimensional, axisymmetric finite element analysis was first used to determine if the results from such an analysis, with emphasis on the shear deformations between the belts, could be used to predict a relative ranking for belt edge endurance. It is shown that such an analysis can lead to erroneous conclusions. A three-dimensional analysis in which tires are modeled under free rotation and static vertical loading was performed next. This approach resulted in an improvement in the quality of the correlations. The differences in the predicted values of various stress analysis parameters for the three belt edge configurations are studied and their implication on predicting belt edge endurance is discussed.


Sign in / Sign up

Export Citation Format

Share Document