scholarly journals An innovative coating technique for light and electron microscopic autoradiography.

1992 ◽  
Vol 40 (6) ◽  
pp. 879-882 ◽  
Author(s):  
G V Kornhauser ◽  
J M Krum ◽  
J M Rosenstein
Keyword(s):  
Background Radiation ◽  
Silver Halide ◽  
Dip Coating ◽  
Electron Microscopic ◽  
Laboratory Procedure ◽  
Electron Microscopic Autoradiography ◽  
Thin Sections ◽  
Coating Technique ◽  
Glass Slides ◽  
Silver Halide Crystals

We describe a modified nuclear emulsion coating technique for both electron and light microscopic autoradiography. We propose that by reversing the application of formvar film so that it adheres to and covers thin sections placed on grids, we have developed a technically accessible methodology that produces optimal conditions for the tracing of specific nuclear activity. A smooth, continuous base is formed over the sections on which a monolayer of evenly packed silver halide crystals can be applied by dip-coating. The same principle is applied to pre-stained 1-micron plastic sections of glass slides. We suggest that the application of formvar film over thin sections does not impede or interfere with the exposure of the emulsion by the labeled tissue. On the contrary, it virtually eliminates contamination and background radiation, enhancing the specificity and quality of resolution at even low magnifications. This technical modification, which facilitates the application of the emulsion, could render electron microscopic autoradiography a routine laboratory procedure, allowing for easily reproducible results and quantitative evaluation. At the light microscopic level, this technique prevents chemical fogging caused by certain stains, and thus allows routine pre-staining before coating with emulsion.

scholarly journals HIGH-RESOLUTION AUTORADIOGRAPHY

1962 ◽  
Vol 15 (2) ◽  
pp. 173-188 ◽  
Author(s):  
Lucien G. Caro ◽  
Robert P. van Tubergen
Keyword(s):  
High Resolution ◽  
Silver Halide ◽  
Electron Microscopic ◽  
Simple Method ◽  
Electron Microscopic Autoradiography ◽  
Fine Grain ◽  
High Resolution Autoradiography ◽  
Silver Halide Crystals ◽  
Subcellular Level ◽  
Physical Developer

Methods used in obtaining high resolution in autoradiography, with special emphasis on the technique of electron microscopic autoradiography, are described, together with control experiments designed to establish the optimum conditions or procedures. On the basis of these experiments the emulsion selected was Ilford L-4, with a crystal size slightly larger than 0.1 micron. It is applied to the specimen in the form of a gelled film consisting of a monolayer of silver halide crystals. Background, when present, can be eradicated by a simple method. The preparations can be stored, in presence of a drying agent, at room temperature or in a refrigerator. Photographic development is done in Microdol, or in a special fine grain "physical" developer. For examination in the electron microscope the sections are stained with uranyl or lead stains. These methods give a good localization of the label, at the subcellular level, and good reproducibility in relative grain counts.

Electron microscopic analysis of objects in light microscopic sections

1978 ◽  
Vol 36 (2) ◽  
pp. 80-81
Author(s):  
Arvid B. Maunsbach
Keyword(s):  
Electron Microscopy ◽  
Light And Electron Microscopy ◽  
Microscopic Analysis ◽  
Electron Microscopic Level ◽  
Semithin Section ◽  
Cover Glass ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Glass Slides ◽  
Ultrathin Sections

Structural studies in experimental biology or in pathology are frequently extended from the light to the electron microscopic level. This is often done by cutting both semithin (about 1 μm) and thin sections from the same tissue block after embedding for electron microscopy. However, in many studies it would be of great value to analyse the same structure both by light and electron microscopy, i.e. to be able to study by electron microscopy an object which is first detected by light microscopy in a semithin section. To achieve this, a method has been developed by which ultrathin sections are cut directly from the semithin section containing the object of interest.Semithin sections, about 1 μ in thickness, are cut from Epon or Vestopal embedded tissue. The sections are placed on ordinary glass slides and stained with toluidine blue. The sections are studied in the light microscope without a cover glass or mounted in water.

Electron-microscopic autoradiography of tritiated testosterone in rat testis

1977 ◽  
Vol 26 (1) ◽  
pp. 339-346
Author(s):  
P.M. Frederick ◽  
H.J. van der Molen ◽  
D. Klepper ◽  
H. Galjaard
Keyword(s):  
Electron Microscope ◽  
Seminiferous Tubules ◽  
Cell Cytoplasm ◽  
Electron Microscopic ◽  
Electron Microscope Level ◽  
Tissue Samples ◽  
Electron Microscopic Autoradiography ◽  
Thin Sections ◽  
Freeze Dried ◽  
Lipid Inclusions

The feasibility of a technique for autoradiography of diffusible substances has been further tested by analysing the localization of steroids in rat testes with the light-and electron-microscope. Testes of rats were perfused with tritiated testosterone (3 min) followed by 15-min perfusion with buffer containing a 100-fold of unlabelled testosterone. Tissue samples were frozen, freeze dried, fixed in osmium vapour and embedded in Epon. To exclude extraction of steroids, contact with water and other solvents was prevented during cutting of thin sections on an ultracryotome and further treatments for autoradiography. Light- and electron-microscopic observations indicate that the highest concentration of labelled testosterone was present within the basal parts of the Sertoli cell cytoplasm and in lipid inclusions of Sertoli cells within the seminiferous tubules. This is the first account of autoradiography of steroids at the electron-microscope level.

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
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
High Resolution ◽  
Ultrastructural Analysis ◽  
Thick Section ◽  
Electron Microscopic ◽  
Electron Microscopic Autoradiography ◽  
Glass Slides

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.

The Optical and Electrical Properties of Antimony-Doped Tin Oxide Transparent Conducting Thin Films Prepared by Sol-Gel Dip-Coating Technique

2007 ◽  
Vol 336-338 ◽  
pp. 754-757
Author(s):  
Dao Li Zhang ◽  
Zhi Bing Deng ◽  
Jian Bing Zhang ◽  
Liang Yan Chen
Keyword(s):  
Thin Films ◽  
Tin Oxide ◽  
Sol Gel ◽  
Visible Region ◽  
Dip Coating ◽  
Rutile Structure ◽  
Coating Technique ◽  
Transparent Conducting ◽  
Glass Slides ◽  
Dip Coating Technique

Antimony-doped tin oxide (ATO) transparent conducting thin films were prepared by sol-gel dip-coating technique in the alcohol solution of metal salts of tin (II) chloride dehydrate and antimony tri-chloride. Usual glass slides (25×76×1mm3) were used as the substrates. As-prepared thin films were dried at temperature of 343K and annealed at temperatures of 673~823K. Their optical properties were analyzed by Hitachi U-3310 spectrophotometer. The good optical transmission of the ATO thin films has been obtained as high as 80%-90% in visible region by the optimization of deposition conditions, but decreased substantially in IR region. From the X-ray diffraction (XRD) measurements, it showed that ATO films had the similar structures with the pure tin oxide films, i.e. tetragonal rutile structure, despite of some rhombic SnO crystals. We analyzed the transmittance in the visible region depending on the vary Sb doped level, temperature, and dip-coating times. The sheet resistance of the investigated thin films was determined by four-probe method, showing that it was about 85-1009/□, which decreased with the increase of antimony doped concentration.

Localization of Gallium-67 in Peritoneal Cells by Electron Microscopic Autoradiography

1973 ◽  
Vol 31 ◽  
pp. 404-405
Author(s):  
D. C. Swartzendruber ◽  
Norma L. Idoyaga-Vargas
Keyword(s):  
Clinical Trials ◽  
Soft Tissue ◽  
Tumor Tissue ◽  
Soft Tissue Tumors ◽  
Clinical Usefulness ◽  
Electron Microscopic ◽  
Normal Cells ◽  
Electron Microscopic Autoradiography ◽  
Peritoneal Cells ◽  
Gallium 67

The radionuclide gallium-67 (67Ga) localizes preferentially but not specifically in many human and experimental soft-tissue tumors. Because of this localization, 67Ga is used in clinical trials to detect humar. cancers by external scintiscanning methods. However, the fact that 67Ga does not localize specifically in tumors requires for its eventual clinical usefulness a fuller understanding of the mechanisms that control its deposition in both malignant and normal cells. We have previously reported that 67Ga localizes in lysosomal-like bodies, notably, although not exclusively, in macrophages of the spocytaneous AKR thymoma. Further studies on the uptake of 67Ga by macrophages are needed to determine whether there are factors related to malignancy that might alter the localization of 67Ga in these cells and thus provide clues to discovering the mechanism of 67Ga localization in tumor tissue.

The Immuno-Electron Microscopic Localization of the Mo-Fe Protein Component of Nitrogenase in Cells of Azotobacter Vinelandii

1973 ◽  
Vol 31 ◽  
pp. 574-575
Author(s):  
J. T. Stasny ◽  
R. C. Burns ◽  
R. W. F. Hardy
Keyword(s):  
Cellular Localization ◽  
Direct Evidence ◽  
Azotobacter Vinelandii ◽  
Intracellular Localization ◽  
Protein Component ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Fe Protein ◽  
Intracytoplasmic Membranes

Structure-functlon studies of biological N2-fixation have correlated the presence of the enzyme nitrogenase with increased numbers of intracytoplasmic membranes in Azotobacter. However no direct evidence has been provided for the internal cellular localization of any nitrogenase. Recent advances concerned with the crystallizatiorTand the electron microscopic characterization of the Mo-Fe protein component of Azotobacter nitrogenase, prompted the use of this purified protein to obtain antibodies (Ab) to be conjugated to electron dense markers for the intracellular localization of the protein by electron microscopy. The present study describes the use of ferritin conjugated to goat antitMo-Fe protein immunoglobulin (IgG) and the observations following its topical application to thin sections of N2-grown Azotobacter.

The High Resolution Appearance of Biological Substrates

1969 ◽  
Vol 27 ◽  
pp. 416-417
Author(s):  
Glen B. Haydon
Keyword(s):  
High Resolution ◽  
Phase Image ◽  
Biological Structure ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Resolution Electron ◽  
Discrete Structures ◽  
Large Phase ◽  
Biological Substrates ◽  
High Resolution Electron Micrographs

High resolution electron microscopic study of negatively stained macromolecules and thin sections of tissue embedded in a variety of media are difficult to interpret because of the superimposed phase image granularity. Although all of the information concerning the biological structure of interest may be present in a defocused electron micrograph, the high contrast of large phase image granules produced by the substrate makes it impossible to distinguish the phase ‘points’ from discrete structures of the same dimensions. Theory predicts the findings; however, it does not allow an appreciation of the actual appearance of the image under various conditions. Therefore, though perhaps trivial, training of the cheapest computer produced by mass labor has been undertaken in order to learn to appreciate the factors which affect the appearance of the background in high resolution electron micrographs.

Determination of the phase structure of polymerblends by electron microscopy

1978 ◽  
Vol 36 (1) ◽  
pp. 492-493
Author(s):  
Dr. G. Kaemof
Keyword(s):  
Screw Extruder ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Single Screw Extruder ◽  
Transmission Electron ◽  
The Matrix ◽  
Twin Screw ◽  
Special Case ◽  
Microscopic Investigations

A mixture of polycarbonate (PC) and styrene-acrylonitrile-copolymer (SAN) represents a very good example for the efficiency of electron microscopic investigations concerning the determination of optimum production procedures for high grade product properties.The following parameters have been varied:components of charge (PC : SAN 50 : 50, 60 : 40, 70 : 30), kind of compounding machine (single screw extruder, twin screw extruder, discontinuous kneader), mass-temperature (lowest and highest possible temperature).The transmission electron microscopic investigations (TEM) were carried out on ultra thin sections, the PC-phase of which was selectively etched by triethylamine.The phase transition (matrix to disperse phase) does not occur - as might be expected - at a PC to SAN ratio of 50 : 50, but at a ratio of 65 : 35. Our results show that the matrix is preferably formed by the components with the lower melting viscosity (in this special case SAN), even at concentrations of less than 50 %.

An Electron Microscopic Study of the Acute Effects of Hypothalamic Releasing Factors on the Anterior Pituitary Gland

1968 ◽  
Vol 26 ◽  
pp. 232-233
Author(s):  
P.W. Coates ◽  
E.A. Ashby ◽  
L. Krulich ◽  
A. Dhariwal ◽  
S. McCann
Keyword(s):  
Anterior Pituitary ◽  
Microscopic Investigation ◽  
Uranyl Acetate ◽  
Electron Microscopic Investigation ◽  
Electron Microscopic ◽  
Tissue Samples ◽  
Thin Sections ◽  
Dense Membrane ◽  
Membrane Bound ◽  
Releasing Factors

The morphologic effects on somatotrophs of crude sheep hypothalamic extract prepared from stalk-median eminence were studied by electron microscopy in conjunction with concurrently run bioassays performed on the same tissue samples taken from young adult male Sherman rats.Groups were divided into uninjected controls and injected experimentals sacrificed at 5', 15', and 30' after injection. Half of each anterior pituitary was prepared for electron microscopic investigation, the other half for bioassay. Fixation using collidine buffered osmium tetroxide was followed by dehydration and embedment in Maraglas. Uranyl acetate and lead citrate were used as stains. Thin sections were examined in a Philips EM 200.Somatotrophs from uninjected controls appeared as described in the literature (Fig. 1). In addition to other components, these cells contained moderate numbers of spherical, electron-dense, membrane-bound granules approximately 350 millicrons in diameter.

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