A Novel Method for Automated Acquisition of Tilt Series for Electron Tomography Based on Pre-Calibration of the Specimen Stage

2000 ◽  
Vol 6 (S2) ◽  
pp. 1148-1149
Author(s):  
U. Ziese ◽  
A.H. Janssen ◽  
T.P. van der Krift ◽  
A.G. van Balen ◽  
W.J. de Ruijter ◽  
...  
Keyword(s):  
High Resolution ◽  
3D Reconstruction ◽  
Cell Biology ◽  
Electron Tomography ◽  
Three Dimensional ◽  
Imaging Method ◽  
3D Images ◽  
Tilt Series ◽  
3D Information ◽  
Structural Arrangements

Electron tomography is a three-dimensional (3D) imaging method with transmission electron microscopy (TEM) that provides high-resolution 3D images of structural arrangements. Conventional TEM images are in first approximation mere 2D-projections of a 3D sample under investigation. With electron tomographya series of images is acquired of a sample that is tilted over a large angular range (±70°) with small angular tilt increments (so called tilt-series). For the subsequent 3D-reconstruction, the images of the tilt series are aligned relative to each other and the 3D-reconstruction is computed. Electron tomography is the only technique that can provide true 3D information with nm-scale resolution of individual and unique samples. For (cell) biology and material science applications the availability of high-resolution 3D images of structural arrangements within individual samples provides unique architectural information that cannot be obtained otherwise. Routine application of electron tomography will comprise a major revolutionary step forward in the characterization of complex materials and cellular arrangements.

Automation of Tilt Series Acquisition in Transmission Electron Microscopy for Applications in Biology and Materials Science

2001 ◽  
Vol 7 (S2) ◽  
pp. 88-89
Author(s):  
Ingo Daberkow ◽  
Bernhard Feja ◽  
Peter Sparlinek ◽  
Hans R. Tietz
Keyword(s):  
3D Reconstruction ◽  
Data Acquisition ◽  
San Francisco ◽  
High Speed ◽  
Materials Science ◽  
Electron Tomography ◽  
Three Dimensional ◽  
Max Planck ◽  
Automatic Data ◽  
Tilt Series

During the last decade, computation of a three-dimensional image from a tilt series (3D reconstruction) has become a well established method, of which a variety of implementations are available. The term “electron tomography” is now generally used for this type of data acquisition and 3D reconstruction. An overview over the techniques involved is given in.With the introduction of micro-processor-controlled TEMs and cooled slow-scan CCD cameras and with the progress in performance of high-speed computers, automation of complex imaging procedures became mainly a task of developing appropriate software, using the control facilities of the microscope. in this way, automated electron tomography was realized in 1990 at the Max- Planck-Institute for Biochemistry in Martinsried, and at about the same time at the University of California in San Francisco (UCSF). New techniques for automatic focusing and alignment, developed somewhat earlier , have been integrated in these automated tomography procedures. in the following we discuss the requirements of automatic data acquisition and the present implementation for several TEMs.

A Novel Method of Data Collection for Automated Electron Tomography Based upon Pre-cal1bration of Image Shifts and Defocus Changes

2001 ◽  
Vol 7 (S2) ◽  
pp. 78-79
Author(s):  
Ulrike Ziese ◽  
Ries Janssen ◽  
Willie Geerts ◽  
Theo van der Krift ◽  
Auke van Balen ◽  
...  
Keyword(s):  
Data Collection ◽  
3D Reconstruction ◽  
Electron Tomography ◽  
Ccd Camera ◽  
Current Status ◽  
Imaging Method ◽  
Major Step ◽  
Experimental Conditions ◽  
Digital Format ◽  
Tilt Series

Electron tomography is a three-dimensional (3D) imaging method with transmission electron microscopy (TEM) that provides high-resolution 3D images of structural arrangements. with electron tomography a series of images is acquired of a sample that is tilted over a large angular range (±70°) with small angular tilt increments. For the 3D-reconstruction, the images of the tilt series are aligned relative to each other and the 3D-reconstruction is computed. Electron tomography is the only technique that can provide 3D information with nm-scale resolution of individual and unique samples. Routine application of electron tomography will comprise a major step forward in the characterization of complex materials and cellular arrangements. When collecting tilt series for electron tomography image shifts and defocus changes have to be corrected for by the human operator. The repetitive correction of these changes is highly time consuming, error prone and very hard to carry out under low-dose imaging conditions.Many practical problems are overcome when electron tomography data collection is performed in an automated fashion. Automation includes the (a) image acquisition on a (digital) CCD camera, which implies that (b) changes in image position and defocus can be detected by on-line image processing and (c) immediately be corrected for by computer control of the microscope, (d) Finally, tilt series are directly available in digital format for subsequent processing. Typically, carrying out such an experiment would take a day, and the actual data collection 2-4 hours. in spite of the enormous progress made in terms of data collection speed during the last few years, the current status of automated tomography still does not meet the requirements that would make it a routinely applicable tool. For a great number of biological assays and research projects, results obtained under different experimental conditions have to be compared, and series of experiments have to be carried out. Therefore, we propose a novel approach for recording a tilt series that significantly increases data collection speed, and widens the applicability of the technique.

scholarly journals Awareness of Cryo Electro-Tomography among Dental Students

2020 ◽  
Vol 11 (SPL3) ◽  
pp. 1050-1053
Author(s):  
Nithyanandham Masilamani ◽  
Dhanraj Ganapathy
Keyword(s):  
College Students ◽  
Electron Tomography ◽  
Three Dimensional ◽  
Image Data ◽  
Dental Students ◽  
Imaging Method ◽  
Cross Sectional ◽  
3D Images ◽  
Cryo Electron Tomography ◽  
Diagnostic Applications

CryoElectronomography (CryoET) is indeed an imaging method used to create high resolution (~1-4 nm) three-dimensional viewpoints of specimen, usually physiological macromolecules as well as cell lines. CryoET is really a highly specialized implementation of scanning electron microscopy cryomicroscopy whereby the specimen are scanned since they are tilted, triggering a series of Image data which can be processed to create a 3d image, analogous to 3D images, similar to a CT scan of the human body. This survey was done for assessing the awareness of Cryo electro tomography amongst dental students. This was a questionnaire oriented cross-sectional type of survey comprising 100 dental college students in Chennai. A self-designed questionnaire comprising ten questions based on the knowledge and awareness aboutCryo-electron tomography amongst dental college students. Questionnaires were circulated through an online website survey planet. The questions explored the awareness of using Cryo-electron tomography as a tool to study various biological applications. After the responses were received from 100 participants, data was collected and analyzed .7% are aware about Cryo Electro-tomography. 3% are aware of the mechanism of action of Cryo Electro-tomography. 5% are aware of the diagnostic applications of Cryo Electro-tomography. 3% are aware of the limitations Cryo Electro-tomography.91% are willing to learn about Cryo Electro-tomography. This study concluded that dental students showed less knowledge and awareness toward Cryo Electro-tomography. There are large gaps in the knowledge and attitudes requiring strong remedial measures.

Embedded Gold Markers for Improved TEM/STEM Tomography Reconstruction

2008 ◽  
Author(s):  
Jian-Shing Luo ◽  
Chia-Chi Huang ◽  
Jeremy D. Russell
Keyword(s):  
3D Reconstruction ◽  
Feature Tracking ◽  
Semiconductor Device ◽  
Electron Tomography ◽  
Tracking Process ◽  
Carbon Coated ◽  
Novel Method ◽  
Tomography Reconstruction ◽  
Tilt Series ◽  
20 Nm

Abstract Electron tomography includes four main steps: tomography data acquisition, image processing, 3D reconstruction, and visualization. After acquisition, tilt-series alignments are performed. Two methods are used to align the tilt-series: cross-correlation and feature tracking. Normally, about 10-20 nm of fiducial markers, such as gold beads, are deposited onto one side of 100 mesh carbon-coated grids during the feature-tracking process. This paper presents a novel method for preparing electron tomography samples with gold beads inside to improve the feature tracking process and quality of 3D reconstruction. Results show that the novel electron tomography sample preparation method improves image alignment, which is essential for successful tomography in many contemporary semiconductor device structures.

scholarly journals High-resolution three-dimensional probes of biomaterials and their interfaces

2012 ◽  
Vol 370 (1963) ◽  
pp. 1337-1351 ◽  
Author(s):  
Kathryn Grandfield ◽  
Anders Palmquist ◽  
Håkan Engqvist
Keyword(s):  
High Resolution ◽  
Electron Tomography ◽  
Three Dimensional ◽  
Controlled Drug Release ◽  
Biological Tissues ◽  
Dimensional Structure ◽  
Bone Interface ◽  
Biomaterial Interfaces ◽  
Titania Coating

Interfacial relationships between biomaterials and tissues strongly influence the success of implant materials and their long-term functionality. Owing to the inhomogeneity of biological tissues at an interface, in particular bone tissue, two-dimensional images often lack detail on the interfacial morphological complexity. Furthermore, the increasing use of nanotechnology in the design and production of biomaterials demands characterization techniques on a similar length scale. Electron tomography (ET) can meet these challenges by enabling high-resolution three-dimensional imaging of biomaterial interfaces. In this article, we review the fundamentals of ET and highlight its recent applications in probing the three-dimensional structure of bioceramics and their interfaces, with particular focus on the hydroxyapatite–bone interface, titanium dioxide–bone interface and a mesoporous titania coating for controlled drug release.

X-ray microtomographic imaging and analysis for basic research

2006 ◽  
Vol 21 (2) ◽  
pp. 125-131 ◽  
Author(s):  
J. H. Dunsmuir ◽  
S. Bennett ◽  
L. Fareria ◽  
A. Mingino ◽  
M. Sansone
Keyword(s):  
Spatial Resolution ◽  
Three Dimensional ◽  
Basic Research ◽  
Integrated Approach ◽  
Imaging Method ◽  
Structure Property ◽  
3D Images ◽  
X Ray ◽  
Structure Property Relationships ◽  
Essential Components

For research facilities with access to synchrotron X-ray sources, X-ray absorption microtomography (XMT) has evolved from an experimental imaging method to a specialized, if not yet routine, microscopy for imaging the three-dimensional (3D) distribution of linear attenuation coefficients and, in some cases, elemental concentration with micron spatial resolution. Recent advances in source and detector design have produced conventional X-ray source instruments with comparable spatial resolution but with lower throughput and without element specific imaging. Both classes of instrument produce 3D images for analysis. We discuss an integrated approach for the implementation of analytical XMT to support basic research into the structure-property relationships of a variety of materials. The essential components include instrumentation for collecting quantitative 3D images, a 3D image processing environment to address questions as to the quantity, composition, geometry, and relationships among the features in one or more images, and visualization to provide insight and communicate results. We give examples of image analysis of resolved and unresolved pore spaces of sandstones.

Visualization of mouse spinal cord intramedullary arteries using phase- and attenuation-contrast tomographic imaging

2016 ◽  
Vol 23 (4) ◽  
pp. 966-974 ◽  
Author(s):  
Yong Cao ◽  
Xianzhen Yin ◽  
Jiwen Zhang ◽  
Tianding Wu ◽  
Dongzhe Li ◽  
...  
Keyword(s):  
Spinal Cord ◽  
High Resolution ◽  
Synchrotron Radiation ◽  
Three Dimensional ◽  
Small Vessel ◽  
Imaging Method ◽  
Central Sulcus ◽  
Contrast Imaging ◽  
Mouse Spinal Cord ◽  
Small Vessels

Many spinal cord circulatory disorders present the substantial involvement of small vessel lesions. The central sulcus arteries supply nutrition to a large part of the spinal cord, and, if not detected early, lesions in the spinal cord will cause irreversible damage to the function of this organ. Thus, early detection of these small vessel lesions could potentially facilitate the effective diagnosis and treatment of these diseases. However, the detection of such small vessels is beyond the capability of current imaging techniques. In this study, an imaging method is proposed and the potential of phase-contrast imaging (PCI)- and attenuation-contrast imaging (ACI)-based synchrotron radiation for high-resolution tomography of intramedullary arteries in mouse spinal cord is validated. The three-dimensional vessel morphology, particularly that of the central sulcus arteries (CSA), detected with these two imaging models was quantitatively analyzed and compared. It was determined that both PCI- and ACI-based synchrotron radiation can be used to visualize the physiological arrangement of the entire intramedullary artery network in the mouse spinal cord in both two dimensions and three dimensions at a high-resolution scale. Additionally, the two-dimensional and three-dimensional vessel morphometric parameter measurements obtained with PCI are similar to the ACI data. Furthermore, PCI allows efficient and direct discrimination of the same branch level of the CSA without contrast agent injection and is expected to provide reliable biological information regarding the intramedullary artery. Compared with ACI, PCI might be a novel imaging method that offers a powerful imaging platform for evaluating pathological changes in small vessels and may also allow better clarification of their role in neurovascular disorders.

Three-dimensional reconstruction based on images from spiral high-resolution computed tomography of the temporal bone: anatomy and clinical application

2005 ◽  
Vol 119 (9) ◽  
pp. 693-698 ◽  
Author(s):  
Beom-Cho Jun ◽  
Sun-Wha Song ◽  
Ju-Eun Cho ◽  
Chan-Soon Park ◽  
Dong-Hee Lee ◽  
...  
Keyword(s):  
Computed Tomography ◽  
High Resolution ◽  
3D Reconstruction ◽  
Temporal Bone ◽  
Three Dimensional ◽  
Image Data ◽  
Computer Assisted ◽  
Surface Rendering ◽  
High Resolution Computed Tomography

The aim of this study was to investigate the usefulness of a three-dimensional (3D) reconstruction of computed tomography (CT) images in determining the anatomy and topographic relationship between various important structures. Using 40 ears from 20 patients with various otological diseases, a 3D reconstruction based on the image data from spiral high-resolution CT was performed by segmentation, volume-rendering and surface-rendering algorithms on a personal computer. The 3D display of the middle and inner ear structures was demonstrated in detail. Computer-assisted measurements, many of which could not be easily measured in vivo, of the reconstructed structures provided accurate anatomic details that improved the surgeon’s understanding of spatial relationships. A 3D reconstruction of temporal bone CT might be useful for education and increasing understanding of the anatomical structures of the temporal bone. However, it will be necessary to confirm the correlation between the 3D reconstructed images and histological sections through a validation study.

Application of electron tomography to elucidate the 3D architecture of diverse asymmetric biological specimens

1993 ◽  
Vol 51 ◽  
pp. 494-495
Author(s):  
Bruce F. McEwen
Keyword(s):  
Electron Tomography ◽  
Tomographic Reconstruction ◽  
Three Dimensional ◽  
Single Copy ◽  
Angular Range ◽  
Electron Dose ◽  
Total Electron ◽  
Reconstruction Methods ◽  
3D Architecture ◽  
3D Information

Electron tomography refers to the use of tomographic reconstruction methods to obtain three-dimensional (3D) information from transmission electron microscopy (TEM). This is accomplished by tilting the sample over a wide angular range with 180° being ideal, but 120° to 140° more typical due to limitations imposed by the geometry of most TEM specimens. If symmetry is present in the sample then the full angular range is not required. Furthermore, if the specimen is present as identical units, either in crystalline arrays or as single particles, the different tilts can be collected from different specimens and hence spare the total electron dose on any given particle. However, many important biological specimens are large, asymmetric objects which, due to their size, are not found as identical individual units. For such cases, the only recourse is to collect all the views from a single copy of the object, generally using a single axis tilt geometry.

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