Ultrastructural Localization of Cholesterol in Human Arteriosclerosis

Author(s):  
Karta Sekhri ◽  
Clint Mills ◽  
Henry Younes

Several reports have appeared recently demonstrating a technique for the localization of cholesterol in a variety of tissues. The technique is based on the premise that free cholesterol reacts with digitonin to form cholesterol digitonide which appears as needle-like crystals or spicules under the electron microscope.In order to study the fine structure of the aorta, surgical biopsies from several patients (ranging in age from 45 to 68 years) and with clinical evidence of arteriosclerosis were fixed promptly in Flickinger's fixative (2% formaldehyde, 2.5% glutaraldehyde) containing 0.2% digitonin but buffered with .1M sodium cacodylate. After fixation for 3 hours aortic samples were washed in cacodylate buffer and post fixed in buffered osmium tetroxide for 2 hours & embedded in Epon-Araldite.

Author(s):  
W.W.K. Cheung ◽  
J.B. Wang

The lantern bug harbours three symbionts, namely a, x and i in its body. These microorganisms are supposed to be transmitted transovarially to the future progeny. The x-symbionts are found in a special pair of organs called the x-organ which bulges to form a rectal organ in adult females. The purpose of this study is to investigate into the fine structure of the x-symbionts. This will serve as a basis for understanding the interactions of this microorganism with its host.The rectum of the lantern bug Pyrops candelaria Linn, was dissected out in buffered insect saline and fixed in 2.5% glutaradehyde in 0.1M sodium cacodylate buffer (pH 7.2) for 1 hr. The rectal organ was subsequently post-fixed in 1% osmium tetroxide (pH 7.2) and dehydrated in alcohol/acetone series. These were blocked in Spurr resin and cut with a Reichert Ultratome. Sections were stained with uranyl acetate and lead citrate and examined with a JEOL JEM-1200EX electron microscope. Thick sections (1 μn) were stained with 1% toluidine blue and examined under a Nikon Optiphot light microscope.


Author(s):  
L. Faso ◽  
E. Rappa ◽  
G. Vernon ◽  
R. Witkus

Although hemocyanin, an oxygen binding protein, is found freely dissolved in the hemolymph of isopods its site of synthesis is still unknown.Circulating blood cel Is such as granular hemocytes have been implicated in hemocyanin synthesis in a number of arthropods including Astacus astacus and Homarus vulgaris. Circulating blood cells of Armadillidium vulgare were examined using a transmission electron microscope (TEM) for evidence of hemocyanin synthesis.For each experiment hemolymph was collected from twenty adult A. vulgare and fixed for 1 hour in 200 uL of 3.5% glutaraldehyde in 0.1M sodium cacodylate buffer pH 7.4 with 0.05% calcium chloride added. Hemolymph was then centrifuged at 3000 rpm in an IEC-DPR-6000 centrifuge for 15 minutes at 15 degrees centigrade. The supernatant was removed, and the resulting pellet was washed with three changes of sodium cacodylate buffer. Postfixation of the pellet was done in 1% osmium tetroxide for 1 hour.


Author(s):  
MB. Tank Buschmann

Development of oligodendrocytes in rat corpus callosum was described as a sequential change in cytoplasmic density which progressed from light to medium to dark (1). In rat optic nerve, changes in cytoplasmic density were not observed, but significant changes in morphology occurred just prior to and during myelination (2). In our study, the ultrastructural development of oligodendrocytes was studied in newborn, 5-, 10-, 15-, 20-day and adult frontal cortex of the golden hamster (Mesocricetus auratus).Young and adult hamster brains were perfused with paraformaldehyde-glutaraldehyde in sodium cacodylate buffer at pH 7.3 according to the method of Peters (3). Tissue samples of layer V of the frontal cortex were post-fixed in 2% osmium tetroxide, dehydrated in acetone and embedded in Epon-Araldite resin.


1978 ◽  
Vol 26 (10) ◽  
pp. 862-864 ◽  
Author(s):  
D M Williams ◽  
J E Linder ◽  
M W Hill ◽  
R Gillett

The ultrastructural localization of alkaline phosphatase in eosinophil leucocytes, obtained from experimentally-induced peritoneal exudates in rats, has been studied using an osmiophilic technique with 2-naphthylthiolphosphoryl dichloride as substrate, fast Blue BBN as diazonium salt and postosmication with 1% aqueous osmium tetroxide. With this method identical incubation procedures could be used for both light and electron microscope examination. Eosinophils were the only cells which contained alkaline phosphatase. The enzyme was predominantly associated with the outer surface of the plasma membrane, being present in much lower concentrations in cytoplasmic cisternae. Eosinophil granules only rarely showed reaction product. The plasma membrane location of alkaline phosphatase in eosinophil leucocytes is identical to that recently demonstrated in the human neutrophil.


1964 ◽  
Vol 23 (2) ◽  
pp. 277-293 ◽  
Author(s):  
Robert M. Herndon

This paper describes the fine structure of the granule cells, stellate neurons, astrocytes, Bergmann glia, oligodendrocytes, and microglia of the rat cerebellum after fixation by perfusion with buffered 1 per cent osmium tetroxide. Criteria are given for differentiating the various cell types, and the findings are correlated with previous light microscope and electron microscope studies of the cerebellum.


1970 ◽  
Vol 16 (11) ◽  
pp. 1041-1044 ◽  
Author(s):  
W. E. McKeen

Osmiophilic bodies appear in parts of the colonial growth of Erysiphe graminis DC. f. sp. hordei Em Marchal culture CR3 growing on the susceptible commercial Keystone variety of barley. They are readily observed by the light and electron microscope after osmium tetroxide staining and are abundant in conidiophores, conidia, and mycelium except in haustorial mother cells, in which they are usually absent. The metabolism of haustorial mother cells is distinct and the fine structure of adjoining cells is frequently different. Osmiophilic bodies are absent from the growing hyphal tip, but gradually increase in number and size further back in the terminal cell. Electron micrographs show that they are intracytoplasmic, intravacuolar, and up to 1 μ in diameter. When the colony is washed with acetone or alcohol rather than with aqueous buffer, after glutaraldehyde fixation, before osmium tetroxide fixation, the osmiophilic bodies are removed, indicating that they are lipids. Fat stains, Sudan black B, and Sudan IV stain these bodies. Perhaps the water needs of the germinating conidium are met in part by the oxidation of fats.


Author(s):  
Deborah A. Kuterbach

Foraging honey bees are believed to use the earth's magnetic field, among other cues, in order to home. It has been reported that the abdomen of the honey bee contains magnetite and iron particles have been localized within abdominal oenocytes. Light microscopic investigations reveal that morphologically detectable iron granules are present only in adult animals older than six days after eclosion (emergence from the comb). This is a report of an ultrastructural examination of the oenocytes during the development of the worker honey bee (Apis mellifera) with particular emphasis on the time of appearance, number, and size of iron granules within the cells.Specimens of the different developmental stages were removed from brood comb, fixed in 2.5% glutaraldehyde in 5mM sodium cacodylate buffer pH 7.3, washed, and post-fixed with 1% osmium tetroxide. In order to preserve the lipid-rich material, rapid dehydration was accomplished by three changes of 50% acetone and two changes of 100% acetone before embedding in Polybed 812 epoxy resin.


1967 ◽  
Vol 2 (2) ◽  
pp. 151-162
Author(s):  
ELIZABETH D. HAY ◽  
J. B. GURDON

Mutant and normal Xenopus laevis embryos (0-nu, 1-nu, 2-nu) were examined in the electron microscope after glutaraldehyde and/or osmium-tetroxide fixation. During cleavage both 0-nu and wild-type embryos contain multiple small nucleolar bodies, less than 1 µ in diameter, composed mainly of a fibrous material. By the end of cleavage or beginning of gastrulation, granular caps develop on the fibrous nucleolar bodies. In 1-and 2-nu cells, the multiple nucleolar bodies are replaced during gastrula and neurula stages by definitive nucleoli (2-5 µ in diameter) which contain abundant small (150 Å) granules intermingled with fibrous material. In 0-nu cells, one or two pseudonucleoli (1-3 µ in diameter) appear at about the same time that definitive nucleoli develop in wild-type cells. The multiple small nucleolar bodies disappear as the pseudonucleoli enlarge. Pseudonucleoli differ from definitive nucleoli in having a much smaller amount of the granular component, which is located as a cap on the periphery of the fibrous component and not mingled with it. The granular component of the 0-nu pseudonucleoli, however, is not distinguishable in its fine structure from the same component of normal nucleoli. In many 0-nu tadpoles at stage 41, the granular component of the nucleolus is entirely absent and the fibrillar component is very prominent. Both granular and fibrous components of the 0-nu pseudonucleoli contain RNA as judged by RNase sensitivity and staining affinity for basic dyes.


1960 ◽  
Vol 7 (3) ◽  
pp. 493-497 ◽  
Author(s):  
Arnaldo Lasansky ◽  
Eduardo de Robertis

The fine structure of the cone and rod outer segments of the toad was studied under the electron microscope after fixation in osmium tetroxide and fixation in formaldehyde followed by chromation. In the OsO4-fixed specimens, the rod outer segment appears to be built of a stack of lobulated flattened sacs, each of which is made of two membranes of about 40 A separated by an innerspace of about 30 A. The distance between the rod sacs is about 50 A. The sacs in the cone outer segment are originated by the folding of a continuous membrane. The thickness of the membranes and width of the spaces between the cone sacs is the same as in rod, but the sac innerspace is slightly narrower in the cone (∼ 20 A). After fixation in formaldehyde and chromation, two different dense lines (l1 and l2) separated by spaces of less density appear. One of the lines, l1, has a thickness of 70 A and is less dense than the other, l2, which is 30 A thick. The correlation of the patterns obtained with both fixatives is considered and two possible interpretations are given. The possibility that l2 is related to a soluble phospholipid component is discussed. It is suggested that the outer segments have a paracrystallin organization similar to that found in myelin.


1963 ◽  
Vol 18 (2) ◽  
pp. 345-365 ◽  
Author(s):  
Peter Satir

Upon excision into spring water, the lateral cilia of the gill of the freshwater mussel Elliptio complanatus (Solander) stop beating, but 0.04 M potassium ion can activate the gill so that these cilia again beat with metachronal rhythm. One per cent osmium tetroxide quickly pipetted onto a fully activated gill fixes the lateral cilia in a pattern that preserves the form and arrangement of the metachronal wave, and permits the cilia to be studied with the electron microscope in all stages of their beat cycle. Changes are seen in the fixed active preparation that are not present in the inactive control, i.e., in the packing of the cilia, the position of the axis of the ciliary cross-section, and the diameter of the ring of peripheral filaments. Analysis of these parameters may lead to new correlations between ciliary fine structure and function.


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