Electron microscopical evaluation of the osmic acid-zinc iodide nerve staining technique (method of Champy-Maillet) on pulmonary tissue

1969 ◽  
Vol 74 (1) ◽  
pp. 125-137 ◽  
Author(s):  
J.M. Lauweryns ◽  
J. Peuskens
Author(s):  
Werner J. Niklowitz

After intoxication of rabbits with certain substances such as convulsant agents (3-acetylpyridine), centrally acting drugs (reserpine), or toxic metal compounds (tetraethyl lead) a significant observation by phase microscope is the loss of contrast of the hippocampal mossy fiber layer. It has been suggested that this alteration, as well as changes seen with the electron microscope in the hippocampal mossy fiber boutons, may be related to a loss of neurotransmitters. The purpose of these experiments was to apply the OsO4-zinc-iodide staining technique to the study of these structural changes since it has been suggested that OsO4-zinc-iodide stain reacts with neurotransmitters (acetylcholine, catecholamines).Domestic New Zealand rabbits (2.5 to 3 kg) were used. Hippocampal tissue was removed from normal and experimental animals treated with 3-acetylpyridine (antimetabolite of nicotinamide), reserpine (anti- hypertensive/tranquilizer), or iproniazid (antidepressant/monamine oxidase inhibitor). After fixation in glutaraldehyde hippocampal tissue was treated with OsO4-zinc-iodide stain and further processed for phase and electron microscope studies.


2010 ◽  
Vol 32 (1) ◽  
pp. 37-46
Author(s):  
원성윤 ◽  
Mi-Sun Hur ◽  
김희진 ◽  
Jongtae Park ◽  
곽현호 ◽  
...  

Author(s):  
J. J. Hebert ◽  
T. P. Hensarling ◽  
T. J. Jacks ◽  
R. J. Berni

It is known that the cellulose layers in cotton fibers are made up of filament-like units termed fibrils. With the advent of electron microscopical techniques, an extensive effort has been put forth to arrive at a size for these units. The currently accepted value for the diameter of these elementary fibrils is approximately 35 A. U.In this report an indirect staining technique for electron microscopy has been employed to study the effect of mercerizing strength sodium hydroxide on the size of the elementary fibrils in cotton fibers. The method involves the addition of an unsaturated carbon-carbon double bond to the cellulose within the fibril and further reacting the double bond with osmium tetroxide. The compound used in this investigation was acrylic acid which not only has the osmium reactive bond but, because of its short molecular length, allows the osmium to situate itself at a close proximity to the structure under study.


Author(s):  
R.W. Horne

The technique of surrounding virus particles with a neutralised electron dense stain was described at the Fourth International Congress on Electron Microscopy, Berlin 1958 (see Home & Brenner, 1960, p. 625). For many years the negative staining technique in one form or another, has been applied to a wide range of biological materials. However, the full potential of the method has only recently been explored following the development and applications of optical diffraction and computer image analytical techniques to electron micrographs (cf. De Hosier & Klug, 1968; Markham 1968; Crowther et al., 1970; Home & Markham, 1973; Klug & Berger, 1974; Crowther & Klug, 1975). These image processing procedures have allowed a more precise and quantitative approach to be made concerning the interpretation, measurement and reconstruction of repeating features in certain biological systems.


Author(s):  
Kosuke Ueda ◽  
Hiroto Washida ◽  
Nakazo Watari

IntroductionHemoglobin crystals in the red blood cells were electronmicroscopically reported by Fawcett in the cat myocardium. In the human, Lessin revealed crystal-containing cells in the periphral blood of hemoglobin C disease patients. We found the hemoglobin crystals and its agglutination in the erythrocytes in the renal cortex of the human renal lithiasis, and these patients had no hematological abnormalities or other diseases out of the renal lithiasis. Hemoglobin crystals in the human erythrocytes were confirmed to be the first case in the kidney.Material and MethodsTen cases of the human renal biopsies were performed on the operations of the seven pyelolithotomies and three ureterolithotomies. The each specimens were primarily fixed in cacodylate buffered 3. 0% glutaraldehyde and post fixed in osmic acid, dehydrated in graded concentrations of ethanol, and then embedded in Epon 812. Ultrathin sections, cut on LKB microtome, were doubly stained with uranyl acetate and lead citrate.


Author(s):  
F.W. Van Leeuwen

In order to obtain specific and optimal ultrastructural localization of vasopressin and oxytocin in the hypothalamo-neurohypophyseal system of the rat, 2 staining procedures and several tissue treatments were evaluated using neurohypophyseal tissue. It appeared from these studies that post-embedding staining with the unlabeled antibody enzyme method developed by Sternberger allows greater dilution of the first antibody (anti-vasopressin, 1:4800) than the indirect procedure (1:320) using a peroxidase conjugate as second antibody. Immersion fixation with 4% formalin during 24 hours gave better results (general ultrastructure, immunoreactivity) than those obtained by perfusion fixation with 2.5% glutaraldehyde-1% paraformaldehyde or freeze substitution.Since no reliable specificity tests were performed at the electron microscopical level, tests were developed for antibodies against both vasopressin and oxytocin. For anti-vasopressin plasma neural lobes of homozygous Brattleboro rats, that are lacking vasopressin by a genet- ical defect, were used. For antibodies against both hormones serial sections were used that were alternately incubated with the antibodies.


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