Detection of Defective Regions in 3D Reconstruction to Support Image Acquisition

A relatively new entry, in the field of microscopy, is the Scanning X-Ray Fluorescence Microscope (SXRFM). Using this type of instrument (e.g. Kevex Omicron X-ray Microprobe), one can obtain multiple elemental x-ray images, from the analysis of materials which show heterogeneity. The SXRFM obtains images by collimating an x-ray beam (e.g. 100 μm diameter), and then scanning the sample with a high-speed x-y stage. To speed up the image acquisition, data is acquired "on-the-fly" by slew-scanning the stage along the x-axis, like a TV or SEM scan. To reduce the overhead from "fly-back," the images can be acquired by bi-directional scanning of the x-axis. This results in very little overhead with the re-positioning of the sample stage. The image acquisition rate is dominated by the x-ray acquisition rate. Therefore, the total x-ray image acquisition rate, using the SXRFM, is very comparable to an SEM. Although the x-ray spatial resolution of the SXRFM is worse than an SEM (say 100 vs. 2 μm), there are several other advantages.

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The IBM-PC in electron microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100165525 ◽

1996 ◽

Vol 54 ◽

pp. 612-613

Author(s):

James F. Mancuso

Keyword(s):

Head Start ◽

Image Acquisition ◽

Cost Effective ◽

Financial Applications ◽

Number Of Factors ◽

Instrument Control ◽

The Cost ◽

Training And Support ◽

Control Image ◽

Ibm Pc

IBM PC compatible computers are widely used in microscopy for applications ranging from control to image acquisition and analysis. The choice of IBM-PC based systems over competing computer platforms can be based on technical merit alone or on a number of factors relating to economics, availability of peripherals, management dictum, or simple personal preference.IBM-PC got a strong “head start” by first dominating clerical, document processing and financial applications. The use of these computers spilled into the laboratory where the DOS based IBM-PC replaced mini-computers. Compared to minicomputer, the PC provided a more for cost-effective platform for applications in numerical analysis, engineering and design, instrument control, image acquisition and image processing. In addition, the sitewide use of a common PC platform could reduce the cost of training and support services relative to cases where many different computer platforms were used. This could be especially true for the microscopists who must use computers in both the laboratory and the office.

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Digital image acquisition, processing, and analysis in a polymer microscopy laboratory

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100170001 ◽

1994 ◽

Vol 52 ◽

pp. 454-455

Author(s):

Vinod K. Berry ◽

Xiao Zhang

Keyword(s):

United States ◽

Image Analysis ◽

Digital Image ◽

Image Acquisition ◽

Image Transmission ◽

The United States ◽

Quantitative Image Analysis ◽

Quantitative Image

In recent years it became apparent that we needed to improve productivity and efficiency in the Microscopy Laboratories in GE Plastics. It was realized that digital image acquisition, archiving, processing, analysis, and transmission over a network would be the best way to achieve this goal. Also, the capabilities of quantitative image analysis, image transmission etc. available with this approach would help us to increase our efficiency. Although the advantages of digital image acquisition, processing, archiving, etc. have been described and are being practiced in many SEM, laboratories, they have not been generally applied in microscopy laboratories (TEM, Optical, SEM and others) and impact on increased productivity has not been yet exploited as well.In order to attain our objective we have acquired a SEMICAPS imaging workstation for each of the GE Plastic sites in the United States. We have integrated the workstation with the microscopes and their peripherals as shown in Figure 1.

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A basic introduction to image processing using NIH-Image as a model

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100169699 ◽

1994 ◽

Vol 52 ◽

pp. 392-393

Author(s):

John Mansfield

Keyword(s):

Image Processing ◽

Digital Image ◽

Image Acquisition ◽

Digital Instrument ◽

Novice User ◽

Nih Image ◽

The Past ◽

Digital World ◽

The World ◽

Instrument Control

Advances in camera technology and digital instrument control have meant that in modern microscopy, the image that was, in the past, typically recorded on a piece of film is now recorded directly into a computer. The transfer of the analog image seen in the microscope to the digitized picture in the computer does not mean, however, that the problems associated with recording images, analyzing them, and preparing them for publication, have all miraculously been solved. The steps involved in the recording an image to film remain largely intact in the digital world. The image is recorded, prepared for measurement in some way, analyzed, and then prepared for presentation.Digital image acquisition schemes are largely the realm of the microscope manufacturers, however, there are also a multitude of “homemade” acquisition systems in microscope laboratories around the world. It is not the mission of this tutorial to deal with the various acquisition systems, but rather to introduce the novice user to rudimentary image processing and measurement.

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Detection, Averaging, and 3D Reconstruction of Biological Specimens on Hypercubes (Transputer-based) Computers

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100181026 ◽

1990 ◽

Vol 48 (1) ◽

pp. 454-455

Author(s):

Jose-Maria Carazo ◽

I. Benavides ◽

S. Marco ◽

J.L. Carrascosa ◽

E.L. Zapata

Keyword(s):

3D Reconstruction ◽

3D Structure ◽

Three Dimensional ◽

Software Tools ◽

Mapping Method ◽

Computer Architectures ◽

Parallel Computer ◽

Reconstruction Process ◽

Computing Power ◽

Order Of Magnitude

Obtaining the three-dimensional (3D) structure of negatively stained biological specimens at a resolution of, typically, 2 - 4 nm is becoming a relatively common practice in an increasing number of laboratories. A combination of new conceptual approaches, new software tools, and faster computers have made this situation possible. However, all these 3D reconstruction processes are quite computer intensive, and the middle term future is full of suggestions entailing an even greater need of computing power. Up to now all published 3D reconstructions in this field have been performed on conventional (sequential) computers, but it is a fact that new parallel computer architectures represent the potential of order-of-magnitude increases in computing power and should, therefore, be considered for their possible application in the most computing intensive tasks.We have studied both shared-memory-based computer architectures, like the BBN Butterfly, and local-memory-based architectures, mainly hypercubes implemented on transputers, where we have used the algorithmic mapping method proposed by Zapata el at. In this work we have developed the basic software tools needed to obtain a 3D reconstruction from non-crystalline specimens (“single particles”) using the so-called Random Conical Tilt Series Method. We start from a pair of images presenting the same field, first tilted (by ≃55°) and then untilted. It is then assumed that we can supply the system with the image of the particle we are looking for (ideally, a 2D average from a previous study) and with a matrix describing the geometrical relationships between the tilted and untilted fields (this step is now accomplished by interactively marking a few pairs of corresponding features in the two fields). From here on the 3D reconstruction process may be run automatically.

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Structural Studies on the Eukaryotic Ribosome

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100180033 ◽

1990 ◽

Vol 48 (1) ◽

pp. 256-257

Author(s):

Adriana Verschoor ◽

Ronald Milligan ◽

Suman Srivastava ◽

Joachim Frank

Keyword(s):

Chick Embryo ◽

3D Reconstruction ◽

Single Particle ◽

Mass Density ◽

Particle Surface ◽

Uranyl Acetate ◽

Electron Microscopic ◽

Eukaryotic Ribosome ◽

Negative Stain ◽

Reconstruction Methods

We have studied the eukaryotic ribosome from two vertebrate species (rabbit reticulocyte and chick embryo ribosomes) in several different electron microscopic preparations (Fig. 1a-d), and we have applied image processing methods to two of the types of images. Reticulocyte ribosomes were examined in both negative stain (0.5% uranyl acetate, in a double-carbon preparation) and frozen hydrated preparation as single-particle specimens. In addition, chick embryo ribosomes in tetrameric and crystalline assemblies in frozen hydrated preparation have been examined. 2D averaging, multivariate statistical analysis, and classification methods have been applied to the negatively stained single-particle micrographs and the frozen hydrated tetramer micrographs to obtain statistically well defined projection images of the ribosome (Fig. 2a,c). 3D reconstruction methods, the random conical reconstruction scheme and weighted back projection, were applied to the negative-stain data, and several closely related reconstructions were obtained. The principal 3D reconstruction (Fig. 2b), which has a resolution of 3.7 nm according to the differential phase residual criterion, can be compared to the images of individual ribosomes in a 2D tetramer average (Fig. 2c) at a similar resolution, and a good agreement of the general morphology and of many of the characteristic features is seen.Both data sets show the ribosome in roughly the same ’view’ or orientation, with respect to the adsorptive surface in the electron microscopic preparation, as judged by the agreement in both the projected form and the distribution of characteristic density features. The negative-stain reconstruction reveals details of the ribosome morphology; the 2D frozen-hydrated average provides projection information on the native mass-density distribution within the structure. The 40S subunit appears to have an elongate core of higher density, while the 60S subunit shows a more complex pattern of dense features, comprising a rather globular core, locally extending close to the particle surface.

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