An Ultrastructural Comparison of Hepatocytes from ADH+ and ADH−Peromyscus Maniculatus

1996 ◽  
Vol 54 ◽  
pp. 30-31
Author(s):  
J. T. Ellzey ◽  
D. Borunda ◽  
B. P. Stewart
Keyword(s):  
South Carolina ◽  
Adult Male ◽  
Uranyl Acetate ◽  
Model System ◽  
Deer Mice ◽  
En Bloc ◽  
Genetic Stock ◽  
Hepes Buffer ◽  
Transmission Electron ◽  
Color Scanner

Genetically alcohol deficient deer mice (ADHN/ADHN) (obtained from the Peromyscus Genetic Stock Center, Univ. of South Carolina) lack hepatic cytosolic alcohol dehydrogenase. In order to determine if these deer mice would provide a model system for an ultrastructural study of the effects of ethanol on hepatocyte organelles, 75 micrographs of ADH+ adult male deer mice (n=5) were compared with 75 micrographs of ADH− adult male deer mice (n=5). A morphometric analysis of mitochondrial and peroxisomal parameters was undertaken.The livers were perfused with 0.1M HEPES buffer followed by 0.25% glutaraldehyde and 2% sucrose in 0.1M HEPES buffer (4C), removed, weighed and fixed by immersion in 2.5% glutaraldehyde in 0.1M HEPES buffer, pH 7.4, followed by a 3,3’ diaminobenzidine (DAB) incubation, postfixation with 2% OsO4, en bloc staining with 1% uranyl acetate in 0.025M maleate-NaOH buffer, dehydrated, embedded in Poly/Bed 812-BDMA epon resin, sectioned and poststained with uranyl acetate and lead citrate. Photographs were taken on a Zeiss EM-10 transmission electron microscope, scanned with a Howtek personal color scanner, analyzed with OPTIMAS 4.02 software on a Gateway2000 4DX2-66V personal computer and stored in Excel 4.0.

Ultrastructural comparison of prolactin adenomas of pituitary in men and women

1988 ◽  
Vol 46 ◽  
pp. 346-347
Author(s):  
R.C. Caughey ◽  
U.P. Kalyan-Raman
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Phosphate Buffer ◽  
Pituitary Adenomas ◽  
Osmium Tetroxide ◽  
Secretory Granules ◽  
Uranyl Acetate ◽  
En Bloc ◽  
Men And Women ◽  
Transmission Electron

Prolactin producing pituitary adenomas are ultrastructurally characterized by secretory granules varying in size (150-300nm), abundance of endoplasmic reticulum, and misplaced exocytosis. They are also subclassified as sparsely or densely granulated according to the amount of granules present. The hormone levels in men and women vary, being higher in men; so also the symptoms vary between both sexes. In order to understand this variation, we studied 21 prolactin producing pituitary adenomas by transmission electron microscope. This was out of a total of 80 pituitary adenomas. There were 6 men and 15 women in this group of 21 prolactinomas.All of the pituitary adenomas were fixed in 2.5% glutaraldehyde, rinsed in Millonig's phosphate buffer, and post fixed with 1% osmium tetroxide. They were then en bloc stained with 0.5% uranyl acetate, rinsed with Walpole's non-phosphate buffer, dehydrated with graded series of ethanols and embedded with Epon 812 epoxy resin.

Microprobe analysis of inclusion bodies related to two benzofuran derivatives

1987 ◽  
Vol 45 ◽  
pp. 672-673
Author(s):  
T. L. Benning ◽  
P. Ingram ◽  
J. D. Shelburne
Keyword(s):  
Inclusion Bodies ◽  
Uranyl Acetate ◽  
Multiple Organ ◽  
High Dose ◽  
En Bloc ◽  
Tissue Samples ◽  
Transmission Electron ◽  
Organ Systems ◽  
Dense Inclusion ◽  
Melanin Complex

Two benzofuran derivatives, chlorpromazine and amiodarone, are known to produce inclusion bodies in human tissues. Prolonged high dose chlorpromazine therapy causes hyperpigmentation of the skin with electron-dense inclusion bodies present in dermal histiocytes and endothelial cells ultrastructurally. The nature of the deposits is not known although a drug-melanin complex has been hypothesized. Amiodarone may also cause cutaneous hyperpigmentation and lamellar lysosomal inclusion bodies have been demonstrated within the cells of multiple organ systems. These lamellar bodies are believed to be the product of an amiodarone-induced phospholipid storage disorder. We performed transmission electron microscopy (TEM) and energy dispersive x-ray microanalysis (EDXA) on tissue samples from patients treated with these drugs, attempting to detect the sulfur atom of chlorpromazine and the iodine atom of amiodarone within their respective inclusion bodies.A skin biopsy from a patient with hyperpigmentation due to prolonged chlorpromazine therapy was fixed in 4% glutaraldehyde and processed without osmium tetroxide or en bloc uranyl acetate for Epon embedding.

Microscopic investigations of novel intracellular bodies of a Nocardia SP

1994 ◽  
Vol 52 ◽  
pp. 356-357
Author(s):  
J. L. Stites
Keyword(s):  
Uranyl Acetate ◽  
Electron Microscopic ◽  
En Bloc ◽  
Ribonucleic Acids ◽  
Transmission Electron ◽  
Nocardia Sp ◽  
Internal Constitution ◽  
Membrane Bound ◽  
Molecular Components ◽  
Protein Rich

A Nocardia sp.was found during an initial transmission electron microscopic (TEM) examination to have unusual intracellular bodies (ICB's) which do not appear to have been described previously in the literature. Most intracellular structures within bacteria have been classified as storage granules, a product of membrane invagination (i.e. mesosomes), or vacuoles. In bacteria there are no known intracellular membrane-bound organelles, and all internal membranes are invaginations of the unit membrane. Several microscopic-level examinations of the Nocardia sp. ICB's were initiated in order to determine their overall structure, classification, and internal constitution.Different TEM staining procedures were performed to determine possible molecular components of the ICB. In all of the staining protocols the ICB's showed a lack of electron density similar to the cell wall. Because the ICB's showed no affinity to any stain, it appeared they do not have strong positive charge (phosphotungstic acid), are not protein rich (en bloc uranyl acetate), lack glycogen and are not phosphate or sulphur rich (lead citrate), nor do they contain lipids or ribonucleic acids (osmium tetroxide).

A Morphometric Analysis of Early Effects of Ethanol on Hepatocyte Organelles from Peromyscusmaniculatus

1998 ◽  
Vol 4 (S2) ◽  
pp. 1064-1065
Author(s):  
J. T. Ellzey ◽  
J. P. Drake ◽  
L. Dader ◽  
P. Boentges
Keyword(s):  
Endoplasmic Reticulum ◽  
South Carolina ◽  
Morphometric Analysis ◽  
Smooth Endoplasmic Reticulum ◽  
Ultrastructural Changes ◽  
Deer Mice ◽  
Pathological Changes ◽  
Genetic Stock ◽  
Early Effects ◽  
Hanta Viruses

Pathological changes of hepatocytes from rats fed a 30% ethanol-derived calories diet for three weeks include noticeable ultrastructural changes including steatosis and hypertrophy of the smooth endoplasmic reticulum. We sought to examine hepatocytes of deer mice administered ethanol in an inhalation chamber for two weeks to determine if subtle changes occur in hepatocyte organelles prior to steatosis.Two strains of Peromyscus maniculatus, ADH-positive possessing hepatic cytosolic alcohol dehydrogenase and ADH-negative deer mice lacking this enzyme were purchased from the Peromyscus Genetic Stock Center (Univ. of South Carolina). They tested negatively for Hanta viruses. A morphometric analysis of the ultrastructure of ADH+(n=14) and ADH- (n=14) controls as well as experimentals exposed to chronic, intoxicating levels of ethanol was conducted. Blood ethanol levels were maintained between 1.25-1.75 mg/ml for two weeks in the experimentals.

Nonspecific structures seen in diagnostic muscle biopsies

1987 ◽  
Vol 45 ◽  
pp. 846-847
Author(s):  
R. C. Caughey ◽  
U. P. Kalyan-Raman
Keyword(s):  
Electron Microscope ◽  
Transmission Electron Microscope ◽  
Osmium Tetroxide ◽  
Clinical History ◽  
Uranyl Acetate ◽  
Ethanol Dehydration ◽  
Tubular Aggregates ◽  
En Bloc ◽  
Transmission Electron ◽  
Muscle Biopsies

In a period of two years we have analyzed 50 muscle biopsies using the transmission electron microscope. Six nonspecific structures consisting of filamentous bodies, tubular aggregates, paracrystalline mitochondrial inclusions, honeycomb arrays, concentric laminated bodies, and finger print profiles were observed in 47 of 50 cases. In order to know the significance of these structures in muscle biopsies, we correlated their occurrence with their clinical history, histological findings, and histochemistry.The biopsies were initially fixed in 2.5% glutaraldehyde (pH. 7.5, 500 mOsm), then randomly minced and post fixed in 1% osmium tetroxide. All biopsies were processed with and without uranyl acetate en bloc staining in Walpole's buffer before ethanol dehydration. They were embedded in Epon 812 epoxy resin, sectioned, and stained with uranyl acetate and lead citrate before evaluation with a JEOL, JEM 100 C Transmission Electron Microscope. All grid squares of six different blocks were scanned to evaluate the ultra-structural pathology.

Electron Microscopy as an aid to diagnosis in pediatric hematology/oncology

1989 ◽  
Vol 47 ◽  
pp. 1062-1063
Author(s):  
K. L. Saving ◽  
R. C. Caughey
Keyword(s):  
Electron Microscopy ◽  
Phosphate Buffer ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
En Bloc ◽  
Thin Sections ◽  
Megakaryoblastic Leukemia ◽  
Pediatric Hematology ◽  
Transmission Electron ◽  
Microscopy Techniques

This presentation is designed to demonstrate how scanning and transmission electron microscopy techniques can be utilized to confirm or support a variety of unusual pediatric hematologic/oncologic disorders. Patients with the following diagnoses will be presented: (1) hereditary pyropoikilocytosis, (2) familial erythrophagocytic lymphohistiocytosis, (3) acute megakaryoblastic leukemia, and (4) pseudo-von Willebrand’s disease.All transmission and scanning electron microscopy samples were fixed in 2.5% glutaraldehyde, rinsed in Millonig’s phosphate buffer, and post-fixed with 1% osmium tetroxide. The transmission samples were then en bloc stained with 0.5% uranyl acetate, rinsed with Walpole ’ s non-phosphate buffer, dehydrated with graded series of ethanols and embedded with Epon 812 epoxy resin. Ultramicrotomy thin sections were stained with uranyl acetate and lead citrate and scanned using a JEOL-JEM 100C, The scanning samples were dehydrated with graded series of ethanols, critical point dried with CO2, gold-coated, and scanned using a JEOL-JSM 35. The peroxidase samples were fixed in 3% glutaraldehyde, incubated in diaminobenzidine (DAB), dehydrated with ethanol, embedded with Epon 812, and scanned without post-staining using a JEOL-JEM 100C.

The ultrastructure of the tegument of adult Schistosoma haematobium

Parasitology ◽  
1984 ◽  
Vol 89 (1) ◽  
pp. 71-78 ◽  
Author(s):  
B. Leitch ◽  
A. J. Probert ◽  
N. W. Runham
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Inclusion Bodies ◽  
Schistosoma Haematobium ◽  
Uranyl Acetate ◽  
En Bloc ◽  
Male Worm ◽  
Elliptical Inclusion ◽  
Transmission Electron ◽  
First Time

SummaryThe ultrastructure of the tegument of Schistosoma haematobium was examined using scanning and transmission electron microscopy. The surface of the male worm is characterized by numerous raised tubercles bearing apically directed spines. The female in contrast to the male is cylindrical and relatively smooth. Details of oral and ventral suckers are given. The use of uranyl acetate as a tertiary fixative and en bloc stain has revealed the heptalaminate nature of the outer membrane. Tegumental mitochondria are shown to be morphologically more complex than those of S. mansoni. Spherical and elliptical inclusion bodies are also described. The ultrastructure of the oesophageal tegument of S. haematobium is described for the first time and corresponds with earlier observations of S. mansoni.

scholarly journals A new fixative for the preservation of actin filaments: fixation of pure actin filament pellets.

1985 ◽  
Vol 33 (11) ◽  
pp. 1116-1128 ◽  
Author(s):  
J Boyles ◽  
L Anderson ◽  
P Hutcherson
Keyword(s):  
Thin Section ◽  
Tannic Acid ◽  
Actin Filaments ◽  
Uranyl Acetate ◽  
Model System ◽  
En Bloc ◽  
Filament Diameter ◽  
Thin Section Microscopy ◽  
Excellent Preservation

Conventional fixation for thin-section microscopy is insufficient to preserve many elements of cells and tissues. Actin filaments, for example, are destroyed during post-fixation in OsO4. In our search for a better fixative, we chose pellets of pure actin filaments as a very sensitive model system. In the present study, the potential of amines for improving aldehyde fixation was explored, and the results were compared to those obtained with the use of tannic acid. Aldehyde and amine were used together as an initial fixative, followed by aldehyde alone with postfixation in 1% OsO4 in buffer at 4 degrees C for 15 min, uranyl acetate en bloc stain, acetone dehydration, and embedding in Epox 812. Some primary monoamines improved the preservation of filaments; filaments were not broken beyond recognition by OsO4, as occurs when glutaraldehyde alone is used. Excellent preservation was seen when certain primary diamines were used. The quality of this fixation was superior to that obtained with tannic acid and was without the large increase in filament diameter that is seen with concentrations of tannic acid sufficient to protect filaments against osmium damage. The effects on filaments of the amines lysine, putrescine, ammonium, and arginine have been documented in detail, as we systematically varied all the major parameters normally considered in formulating fixation protocols.

scholarly journals UA-Zero as a Uranyl Acetate Replacement When Diagnosing Primary Ciliary Dyskinesia by Transmission Electron Microscopy

Diagnostics ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1063
Author(s):  
Andreia Lucia Pinto ◽  
Ranjit Kaur Rai ◽  
Amelia Shoemark ◽  
Claire Hogg ◽  
Thomas Burgoyne
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Cross Sections ◽  
Primary Ciliary Dyskinesia ◽  
Uranyl Acetate ◽  
Lung Damage ◽  
En Bloc ◽  
Suitable Alternative ◽  
Transmission Electron ◽  
Ciliary Dyskinesia

Primary ciliary dyskinesia (PCD) is a disorder affecting motile cilia. An early accurate diagnosis helps prevent lung damage and preserve lung function. To make a diagnostic assessment, one of the commonly used methods that allows for the examination of ciliary ultrastructure is transmission electron microscopy (TEM). This allows for a quantitative assessment of ciliary components to identify defects associated with PCD. Heavy metal staining is required to provide a contrast when imaging cilia in the TEM. One of the most commonly used stains is uranyl acetate (UA). UA can be applied to cellular material before embedding (en bloc), or to ultrathin sections of embedded samples (grid staining). UA is radioactive and, due to growing safety concerns and restrictions by government bodies, universities and hospitals, it is essential to find a suitable alternative. We show UA-zero (UAZ), when used en bloc, provides a high contrast and is a suitable replacement for UA. PCD diagnostic experts, having reviewed ciliary cross-sections stained with UAZ en bloc, are confident that the staining and PCD defects are readily detectable similar to samples that have been stained with UA.

Ultrastructural observations in two cases of acute pyelonephritis

1988 ◽  
Vol 46 ◽  
pp. 364-365
Author(s):  
R.C. Caughey ◽  
C.E. Kelly
Keyword(s):  
Basement Membrane ◽  
Acute Pyelonephritis ◽  
Bacterial Adhesion ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
En Bloc ◽  
Thin Sections ◽  
Transmission Electron ◽  
Tubular Lumen ◽  
Release Characteristic

In two histologically diagnosed cases of acute pyelonephritis, several ultrastructural features were observed. The transmission electron microscope (TEM) highlights the bacterial adhesion, inflammatory process, phagocytosis and lysosomal release characteristic of pyelonephritis.The renal biopsies from both patients were fixed in 2.5% glutaraldehyde, rinsed with Millonig's buffer, and post fixed with 1% osmium tetroxide. They were then en bloc stained with 0.5% uranyl acetate, rinsed with Walpole's buffer, dehydrated with a graded series of ethanols and embedded with Epon 812 epoxy resin. Ultramicrotomy thin sections were stained with uranyl acetate and lead citrate and scanned using a JEOL - JEM100C.Using the TEM, bacteria are seen in tubular epithelial cells, phagocytized to the basement membrane, within cells of the tubular lumen, and within the polymorphonuclear leukocytes (polys). Bacteria may also be seen phagocytizing within cells in the tubular lumen. The inflammatory response of polys is seen in the tubular lumen and interstitium, and attached and within the tubular basement membrane.

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