Platinum Compounds as Stains for Electron Microscopy

1974 ◽  
Vol 32 ◽  
pp. 230-231
Author(s):  
S. K. Aggarwal ◽  
P. McAllister ◽  
R. W. Wagner ◽  
B. Rosenberg
Keyword(s):  
Electron Microscopy ◽  
Hela Cells ◽  
Uranyl Acetate ◽  
Sarcoma 180 ◽  
Platinum Compounds ◽  
Kb Cells ◽  
En Bloc ◽  
Human Lymphoma ◽  
Ultrathin Sections ◽  
Blue Coloration

Uranyl acetate has been used as an electron stain for en bloc staining as well as for staining ultrathin sections in conjunction with various lead stains (Fig. 1). Present studies reveal that various platinum compounds also show promise as electron stains. Certain platinum compounds have been shown to be effective anti-tumor agents. Of particular interest are the compounds with either uracil or thymine as one of the ligands (cis-Pt(II)-uracil; cis-Pt(II)-thymine). These compounds are amorphous, highly soluble in water and often exhibit an intense blue coloration. These compounds show enough electron density to be used as stains for electron microscopy. Most of the studies are based on various cell lines (human AV, cells, human lymphoma cells, KB cells, Sarcoma-180 ascites cells, chick fibroblasts and HeLa cells) while studies on tissue blocks are in progress.

Uranyl Acetate as a Fixative—from pH 2.0 to 8.0

1971 ◽  
Vol 29 ◽  
pp. 490-491
Author(s):  
V. R. Mumaw ◽  
B. L. Munger
Keyword(s):  
Electron Microscopy ◽  
Oxalic Acid ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
En Bloc ◽  
Tissue Sections ◽  
Normal Rat ◽  
Lead Hydroxide ◽  
Ultrathin Sections ◽  
Rat Tissue

Numerous applications utilizing uranyl acetate as an electron stain for electron microscopy have been described. Uranyl acetate has become a routine stain used in conjunction with lead hydroxide for staining ultrathin sections. En bloc staining with uranyl acetate following osmium tetroxide post-fixation produces undesirable effects on some cytoplasmic components, especially glycogen. Recent studies using uranyl acetate as a fixative and en bloc stain at pH 7.2 before osmification has shown uranyl acetate to have desirable fixation and staining qualities. Tissues treated with uranyl acetate at a pH of 2.0-8.0 were studied. Normal rat tissue was fixed in Karnovsky's paraformaldehyde-glutaraldehyde fixative. The tissue was post-fixed in 0.5% uranyl acetate in water at pH 2.0 and 0.5% uranyl acetate in 0.1M s-collidine with 0.01M oxalic acid at pH 4, pH 6.0, pH 7.2, and pH 8.0 for 1 hour at 4°C. Following several rinses of 0.1M s-collidine buffer, the tissues were treated with 1.33% osmium tetroxide 1 hour at 4°C followed by rapid dehydration in ethanol and embedded in Durcupan ACM. Tissue sections were stained with lead hydroxide.

The Ultrastructure of Lymphocytic Hypophysitis

1981 ◽  
Vol 39 ◽  
pp. 466-467
Author(s):  
S.L. Asa ◽  
K. Kovacs ◽  
J. M. Bilbao ◽  
R. G. Josse ◽  
K. Kreines
Keyword(s):  
Electron Microscopy ◽  
Plasma Cells ◽  
Secretory Granules ◽  
Sella Turcica ◽  
Uranyl Acetate ◽  
Lymphocytic Hypophysitis ◽  
Pituitary Cells ◽  
Transmission Electron ◽  
Ultrathin Sections ◽  
Mass Lesions

Seven cases of lymphocytic hypophysitis in women have been reported previously in association with various degrees of hypopituitarism. We report two pregnant patients who presented with mass lesions of the sella turcica, clinically mimicking pituitary adenoma. However, pathologic examination revealed extensive infiltration of the anterior pituitary by lymphocytes and plasma cells with destruction of the gland. To our knowledge, the ultrastructural features of lymphocytic hypophysitis have not been studied so far.For transmission electron microscopy, tissue from surgical specimens was fixed in glutaraldehyde, postfixed in OsO4, dehydrated and embedded in epoxy-resin. Ultrathin sections were stained with uranyl acetate and lead citrate and examined with a Philips 300 electron microscope.Electron microscopy revealed adenohypophysial cells of all types exhibiting varying degrees of injury. In the areas of most dense inflammatory cell infiltration pituitary cells contained large lysosomal bodies fusing with secretory granules (Fig. 1), as well as increased numbers of swollen mitochondria, indicating oncocytic transformation (Fig. 2).

An ultrastructural study of sertoli cells in two geographically isolated populations of Aphanius dispar

1992 ◽  
Vol 50 (1) ◽  
pp. 548-549
Author(s):  
Taber A. Ba-Omar ◽  
Philip F. Prentis
Keyword(s):  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Freshwater Teleost ◽  
Isolated Populations ◽  
En Bloc ◽  
The North ◽  
Group A ◽  
Ultrathin Sections ◽  
Group B ◽  
Geographically Isolated

We have recently carried out a study of spermiogenic differentiation in two geographically isolated populations of Aphanius dispar (freshwater teleost), with a view to ascertaining variation at the ultrastructural level. The sampling areas were the Jebel Al Akhdar in the north (Group A) and the Dhofar region (Group B) in the south. Specimens from each group were collected, the testes removed, fixed in Karnovsky solution, post fixed in OsO, en bloc stained with uranyl acetate and then routinely processed to Agar 100 resin, semi and ultrathin sections were prepared for study.

scholarly journals FINE STRUCTURES OF INTRACYTOPLASMIC ORGANELLES OF MYCOBACTERIA

1961 ◽  
Vol 9 (3) ◽  
pp. 597-608 ◽  
Author(s):  
Masaatsu Koike ◽  
Kenji Takeya
Keyword(s):  
Electron Microscopy ◽  
Lamellar Structure ◽  
Cytoplasmic Membrane ◽  
Uranyl Acetate ◽  
Low Density ◽  
Nuclear Region ◽  
Unit Membrane ◽  
Veronal Buffer ◽  
Ultrathin Sections ◽  
Fine Structures

The fine structure of the intracytoplasmic organelles of mycobacteria was studied by means of electron microscopy of ultrathin sections. A well-preserved nuclear apparatus was obtained by fixation with OsO4 in acetate-veronal buffer, containing calcium and tryptone, or in collidine-HCl buffer, followed by uranyl-acetate treatment and embedding in araldite. A low density nuclear region was filled with fine fibrils, 30 A in diameter, in parallel or concentric arrangement. A membranous organelle, tentatively designated as "lamellar structure," consists of unit membranes in lamellar arrangement. The thickness of each lamella in this membranous organelle coincides with that of the three-layered cytoplasmic membrane Moreover, the continuity of this unit membrane with the cytoplasmic membrane was demonstrated.

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.

Permeability in the Blood-Brain Barrier in Monkeys Following Exposure to High Altitude

1971 ◽  
Vol 29 ◽  
pp. 328-329
Author(s):  
R.S. Demaree ◽  
L.J. Ackerman ◽  
D. L. Anderson
Keyword(s):  
Electron Microscopy ◽  
Cerebral Edema ◽  
Osmium Tetroxide ◽  
Acute Mountain Sickness ◽  
Cebus Apella ◽  
Uranyl Acetate ◽  
Lead Citrate ◽  
Limited Evidence ◽  
Ultrathin Sections ◽  
Mountain Sickness

People who rapidly ascend to high terrestrial elevations may experience the “acute mountain sickness” syndrome. Speculation and limited evidence suggest that cerebral edema may play an important role in initiating and perpetuating this condition. We have recently demonstrated by electron microscopy that a mild cerebral edema develops in some Cebus apella monkeys rapidly transported to 14,110 feet. In the present study, Cebus apella monkeys were terminated at 1, 3, or 5 days after being shipped from sea level (160 feet) to 14,110 feet without acclimatization at intermediate altitudes.Thorotrast was administered IV 30 minutes prior to termination by perfusion or guillotine. Cerebral cortex was fixed by either perfusion or immersion in glutaraldehyde, and postfixed in osmium tetroxide. Following fixation, the tissues were dehydrated in ascending concentrations of ethanol followed by propylene oxide and embedded in Epon 812. Ultrathin sections were either not stained or doubly stained with uranyl acetate and lead citrate.

High-Resolution Stereo Electron Microscopy of Neurofilaments and Alzheimer Type Paired Helical Filaments

1985 ◽  
Vol 43 ◽  
pp. 730-733
Author(s):  
H.M. Wisniewski ◽  
G.Y. Wen
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Water Surface ◽  
Paired Helical Filaments ◽  
Uranyl Acetate ◽  
Saturated Solution ◽  
New Method ◽  
Alzheimer Type ◽  
Tissue Block ◽  
Ultrathin Sections

To learn about the ultrastructural similarities and dissimilarities between the Alzheimer's paired helical filaments (PHF) and normal neurofilaments, we developed a new method to cut the plastic embedded block. We call the sections cut by this new method supra-ultrathin sections. To obtain supra-ultrathin EM sections, a selected area of the tissue block was first trimmed to a size about 0.25 mm2 to reduce the sectioning pressure between the knife and the tissue block. The tissue was sectioned as thin as possible. The interference color of the sections was almost as transparent as the water surface. The thicker sections with different interference colors were removed by picking up with hair tip so that all of the sections floating on the water were of same or similar thickness. A saturated solution of uranyl acetate in 50% ethanol was used to maximize the contrast of these supra-ultrathin EM sections on uncoated grids.

Organs of a Baikal seal affected by a morbillivirus

1990 ◽  
Vol 48 (3) ◽  
pp. 968-969
Author(s):  
O.I. Belykh ◽  
Ye.V. Likhoshway ◽  
Yu.V. Solodun ◽  
O.A. Goldberg ◽  
V.P. Kumarev
Keyword(s):  
Electron Microscopy ◽  
Measles Virus ◽  
Present Report ◽  
Protein A ◽  
Uranyl Acetate ◽  
Kidney Cells ◽  
Electron Microscopic ◽  
Microscopic Studies ◽  
Liver And Kidney ◽  
Ultrathin Sections

The population of Baikal seals Phoca sibirica has been plagued in 1987-88 by an unknown disease. Oligonucleotide probing of nucleic acids isolated from tissues of ill and dead animals, as well as immunological evidence and clinical data suggested that seals were infected by a morbillivirus. Morbillivirus antigen has been vizualized in dead seal tissues by immunoelectron microscopy (preembedding technique).The present report gives outline of electron microscopic studies of the tissues of infected Baikal seals. Morbillivirus antigens were vizualized as clusters of gold spheres by postembedding technique with monoclonal antibodies against measles virus and protein A-colloid gold conjugates in nuclei and cytoplasm of liver and kidney cells. Some clusters were associated with virus-like particles having a diameter of 80-100 nm. Electron microscopy of ultrathin sections stained with uranyl acetate revealed nucleocapsides having length of up to 1400 nm, and a diameter of 13-17 nm, morphologically similar to measles and seals distemper virus.

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