scholarly journals Back to back autoradiography: a method for ultrastructural analysis of light microscopic autoradiographs.

1980 ◽  
Vol 28 (3) ◽  
pp. 276-278 ◽  
Author(s):  
N B Kaplan ◽  
L J Smith ◽  
J S Brody

A method for studying radiolabeled cells is described that combines the simplicity of light microscopic autoradiography with the high resolution of electron microscopy. Serial thin (600 A) and thick (1 micron) sections are placed on Formvar-coated slot grids and glass slides, respectively. Labeled cells are visualized on the thick section by autoradiography and may then be studied in the electron microscope by locating the corresponding fields on the grid. This technique permits accurate ultrastructural identification and analysis of radiolabeled cells, yet avoids the need for electron microscopic autoradiography.

Author(s):  
J. A. Pollock ◽  
M. Martone ◽  
T. Deerinck ◽  
M. H. Ellisman

Localization of specific proteins in cells by both light and electron microscopy has been facilitate by the availability of antibodies that recognize unique features of these proteins. High resolution localization studies conducted over the last 25 years have allowed biologists to study the synthesis, translocation and ultimate functional sites for many important classes of proteins. Recently, recombinant DNA techniques in molecular biology have allowed the production of specific probes for localization of nucleic acids by “in situ” hybridization. The availability of these probes potentially opens a new set of questions to experimental investigation regarding the subcellular distribution of specific DNA's and RNA's. Nucleic acids have a much lower “copy number” per cell than a typical protein, ranging from one copy to perhaps several thousand. Therefore, sensitive, high resolution techniques are required. There are several reasons why Intermediate Voltage Electron Microscopy (IVEM) and High Voltage Electron Microscopy (HVEM) are most useful for localization of nucleic acids in situ.


Author(s):  
J. P. Langmore ◽  
N. R. Cozzarelli ◽  
A. V. Crewe

A system has been developed to allow highly specific derivatization of the thymine bases of DNA with mercurial compounds wich should be visible in the high resolution scanning electron microscope. Three problems must be completely solved before this staining system will be useful for base sequencing by electron microscopy: 1) the staining must be shown to be highly specific for one base, 2) the stained DNA must remain intact in a high vacuum on a thin support film suitable for microscopy, 3) the arrangement of heavy atoms on the DNA must be determined by the elastic scattering of electrons in the microscope without loss or large movement of heavy atoms.


Author(s):  
David Joy ◽  
James Pawley

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. The spatial resolution of images made using such a process 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 point. A third limitation emerges from the fact that the probing beam is composed of a finite 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).


Clay Minerals ◽  
1985 ◽  
Vol 20 (2) ◽  
pp. 255-262 ◽  
Author(s):  
S. Mann ◽  
R. M. Cornell ◽  
U. Schwertmann

Aluminium-substituted goethites are found in many soils and can also be synthesised readily in the laboratory. In recent years, synthetic substituted goethites have been examined by various techniques including XRD, IR, TEM and dissolution kinetics (Thiel, 1963; Jonas & Solymar, 1970; Fey & Dixon, 1981; Fysh & Fredericks, 1983; Schulze & Schwertmann, 1984; Schwertmann, 1984). Transmission electron microscopy (TEM) studies have shown that as Al substitution rises above 10%, the goethite needles become shorter and also thicker in the a direction. Furthermore, crystals which at zero substitution consist of domains parallel to the c axis become less domainic with increasing Al substitution (Schulze & Schwertmann, 1984).


1997 ◽  
Vol 3 (S2) ◽  
pp. 353-354
Author(s):  
William R. McManus ◽  
Donald J. McMahon

Models for the structure of the bovine casein micelle have been proposed in the past (1,2,3,4). These models are based on the chemical and physical properties of the micelles and fall into two general catagories, framework and submicelle models. Electron Microscopy techniques were one of the tools used in the development of these models. However, no definative model has been established. We have developed a new electron microscopy method and it is being applied to the re-examination of the protein structure of the casein micelle, with the goal of establishing a definative model for the structure of the casein micelle.High resolution scannning electron microscopy has proven unsuccessful in revealing the protein structure in the casein micelle (Fig 1). To create high resolution scanning electron microscopic images, fine metal coatings are required in the 5 to l0nm range. Following the application of these coatings, images of casein micelles resemble a melting lumpy sphere. This is due to the metal layer obscuring the proteins from view.


1999 ◽  
Vol 5 (S2) ◽  
pp. 526-527
Author(s):  
Maryann E. Martone

One class of biological structures that has always presented special difficulties to scientists interested in quantitative analysis is comprised of extended structures that possess fine structural features. Examples of these structures include neuronal spiny dendrites and organelles such as the Golgi apparatus and endoplasmic reticulum. Such structures may extend 10's or even 100's of microns, a size range best visualized with the light microscope, yet possess fine structural detail on the order of nanometers that require the electron microscope to resolve. Quantitative information, such as surface area, volume and the micro-distribution of cellular constituents, is often required for the development of accurate structural models of cells and organelle systems and for assessing and characterizing changes due to experimental manipulation. Performing estimates of such quantities from light microscopic data can result in gross inaccuracies because the contribution to total morphometries of delicate features such as membrane undulations and excrescences can be quite significant. For example, in a recent study by Shoop et al, electron microscopic analysis of cultured chick ciliary ganglion neurons showed that spiny projections from the plasmalemma that were not well resolved in the light microscope effectively doubled the surface area of these neurons.While the resolution provided by the electron microscope has yet to be matched or replaced by light microscopic methods, one drawback of electron microscopic analysis has always been the relatively small sample size and limited 3D information that can be obtained from samples prepared for conventional transmission electron microscopy. Reconstruction from serial electron micrographs has provided one way to circumvent this latter problem, but remains one of the most technically demanding skills in electron microscopy. Another approach to 3D electron microscopic imaging is high voltage electron microscopy (HVEM). The greater accelerating voltages of HVEM's allows for the use of much thicker specimens than conventional transmission electron microscopes.


1980 ◽  
Vol 88 (3) ◽  
pp. 252-256 ◽  
Author(s):  
Douglas E. Mattox

The single most important factor determining the patency of a microvascular anastomosis is the surgical precision with which it is performed. Inaccurately placed sutures, damage of the intima, exposed media and adventitia, and stenosis of the lumen at the site anastomosis all contribute to decreased patency rates. The first 50 consecutive microvascular anastomoses performed by a single microvascular surgeon were analyzed in vivo and with the scanning electron microscope. The frequency and significance of various technical errors are discussed. Scanning electron microscopy is recommended as a convenient and quick technique for assessing the evenness and accuracy of intimal apposition in microvascular anastomosis.


1987 ◽  
Vol 111 ◽  
Author(s):  
D. R. Acosta ◽  
O. Guzman ◽  
P. Del Angel ◽  
J. Dominguez

High resolution electron microscopy has proven to be a powerful technique to determine structural characteristics of zeolites (l–2),symmetry variations and identification of several kind of defects.Together with ideal projected potential images, the microscopist usually finds in electron micrographs the influence of electro-optical parameters and alterations of the crystallinity of the material under electron irradiation. One of the purposes of this workis to contributetothe understanding of the degradation process of zeolites under electron irradiation in the electron microscope and in this way, discriminate when it is possible, what is reliable information recorded in the images obtained in high resolution conditions.


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