Ultrastructure of Angiosarcoma of Mouse Tail

Author(s):  
White Yvonne ◽  
Winslow Sheldon ◽  
James W. Townsend ◽  
Neil A. Littlefield

Spontaneous neoplasms rarely occur on the tails of BALB/cStCrlfC3H/Nctr mice, but the neoplasm most frequently observed is a locally invasive non-metastatic angiosarcoma. In this case a female mouse weighing 33.2 g, 706 days of age, presented a soft, red, irregularly-shaped mass, measuring 18 mm in its greatest dimension, in the subcutis of the base of the tail. A portion of the tail tumor was taken for electron microscopy and the remainder was processed for light microscopy. The tissue processed for electron microscopy was fixed in 4% cacodyl ate-buffered glutaraldehyde, post-fixed in 1% osmium tetroxide, dehydrated in a graded series of ethanol solutions, and embedded in Epon-Araldite resin mixture. Sections of 1 μm were stained with toluidine blue for light microscopy and ultrathin sections of 100 nm were stained with uranyl acetate and lead citrate, then examined with a Philips EM201 electron microscope .

Author(s):  
Masami Hokano ◽  
Tsunao Oh-I ◽  
Yoshie Narita ◽  
Hiroshi Sassa ◽  
Saburo Suzuki

Malacoplakia is a kind of granulomatous inflammation and characterized by the presence of calcium-stain positive granules (Michaelis-Gutmann bodies, hereinafter abbreviated as M-G bodies ) in the macrophages.In this report we want to say about the following articles:the ultrastruc tural findings in four cases of vesical malacoplakia;the ultrastructural morphogenesis of the M-G bodies;X-ray microanalytical studies which examine the change of the chemical component of M-G bodies, according to their developing stages; andacid phosphatase activity of lysosome within the malacoplakic macrophages.Biopsy materials taken from four cases with vesical malacoplakia were divided into 2 parts; one for a light microscopy and the other for an electron microscopy. The specimen for an electron microscopy was fixed in glutaraldehyde and osmium tetroxide, dehydrated in ethanol and then embedded in Epon 812. Ultrathin sections were doublestained with lead citrate and uranyl acetate and then subjected to routin TEM observation. For X-ray microanalysis was mounted an energy dispersive X-ray microanalyzer on a JEM-100 C type electron microscope. Ultrathin sections for microanalysis were cut 100-200nm thich and not stained.


Author(s):  
Annette M. Andrews ◽  
Alex M. Cameron ◽  
James W. Townsend ◽  
Winslow G. Sheldon

Pituitaries of 6 C3H/HEJ MTV+ mice about 660 days of age were fixed by immersion in cacodylate-buffered 4% glutaraldehyde, post-fixed in 1% osmium tetroxide, stained en block with aqueous uranyl acetate, dehydrated in a graded series of ethanol solutions, cleared in acetone, and embedded in an Epon-Araldite resin mixture. Semi-thin (1 μm) sections were taken of the entire face of the block, and stained with toluidine blue. Subsequently, thin sections (100 nm) were prepared from mesas of the two sides of the pars distal is and one mesa of the pars intermedia. Thin sections were stained with ethanolic uranyl acetate followed by Sato's lead citrate (1), then examined on a Philips EM201 electron microscope.


Author(s):  
S. R. Bawa ◽  
R. Bawa ◽  
H. K. Bains

Examination of ultrathin sections of the spermatozoa recovered from the epididymis of the Indian palm squirrel (Funambulus penanti) indicates that the sperm head undergoes changes in its curvature during epididymal transit.Testis and epididymis of an adult male squirrel were dissected and small pieces of tissue fixed in 2.0% glutaraldehyde in 0.1 M phosphate buffer and post-fixed in osmium tetroxide. After dehydration in graded acetone the material was embedded in Araldite. Ultrathin sections were cut on a Reichert Jung Ultracut, picked-up on copper grids, stained with Reynold’s lead citrate-uranyl acetate and examined with a JEOL 1200 EX transmission electron microscope.Ultrathin sections of the caput epididymal spermatozoa reveal that their plasma membrane is adherent to the underlying acrosome (Figure 1). When these spermatozoa reach the corpus epididymis the plasma membrane surrounding the head becomes ruffled (Figure 2). The lifting-up of the plasma membrane around the head is restricted to the posterior bend of the acrosome.


Author(s):  
R.S. Demaree ◽  
L.J. Ackerman ◽  
D. L. Anderson

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.


Author(s):  
Daniel C. Pease

It is reasonable to think that phospholipid micelles should be visible and identifiable in electron micrographs of ultrathin sections if only they can be preserved throughout the embedding process. The development of highly polar, water-containing, aminoplastic embedments has made this a likely possibility. With this in mind, an investigation of the lecithin-secreting, Type II pneumocytes of the lung is underway.Initially it has been easiest to recognize phospholipid micelles in lung tissue fixed first with glutaraldehyde, and then secondarily exposed to osmium tetroxide. However, the latter is not a necessary concomitant for micellar preservation. Conventional uranyl acetate and lead citrate staining is finally applied. Importantly, though, the micelles have been most easily seen in tissue embedded in 507. glutaraldehyde polymerized with urea, as described in detail by D.C. Pease and R.G. Peterson (J. Ultra- struct. Res., 41, 133, 1972). When oriented appropriately, the micellar units are seen as tiny, bilayer plates.


Author(s):  
John H. L. Watson ◽  
Jessica Goodwin ◽  
E. Osborne Coates

Biopsies of lung were taken at operation from a patient with semi-acute diffuse pulmonary infiltrates for study by TEM and SEM. Tissue by light microscopy showed non-caseating granulomas consistent with sarcoidosis. Clinical evidence suggested a hypersensitivity reaction related to inhalation of substance of undetermined nature. Samples were fixed in glutaraldehyde, cacodylate-buffered. They were critical point dried and coated with Au-Pd for SEM, and were handled appropriately for TEM in Araldite. Sections were contrasted with uranyl acetate and lead citrate.


Author(s):  
F. A. Al-Bagdadi ◽  
C. W. Titkemeyer ◽  
J. E. Lovell

Skin biopsy samples were collected monthly from the lateral sides of 9 male Beagle dogs over a period of 1 year. The samples were fixed in 3% gluteral- dehyde and post fixed in 1% osmium tetroxide using phosphate or S-Collidine as buffers. They were dehydrated, embedded in Epon 812, sectioned with an LKB Ultrotome III, and stained with Reynolds' lead citrate and 1% uranyl acetate. They were examined and photographed by use of an RCA-3H electron microscope.The dermal papilla during anagen consisted of fibroblast-like cells some of which had cytoplasmic processes. The cytoplasm contained many mitochondria. (Fig. 1) The cells of the dermal papilla were either peripherally arranged spindle-shaped cells or polygonal cells. The basement membrane was either related to the fibroblast cytoplasmic processes or was free of the cytoplasmic processes but with a thick zone composed of fibers, granular ground substance and spaces.


Author(s):  
C.N. Sun ◽  
H.J. White ◽  
R.C. Read

Previously we have reported the defect of collagen fibrils from herniated rectus sheath. This presentation includes additional sections from postsurgical incisions (10 days) from both control and hernia patients. Small pieces of rectus sheath were fixed in 3% glutaraldehyde in phosphate buffer (pH 7.2) and post fixed with buffered 2% osmium tetroxide. The tissues were then dehydrated in serially increasing concentrations of alcohol and embedded in Epon 812. Sections were stained with 2.5% phosphotungstic acid or uranyl acetate and lead citrate.Previously we found that collagen fibrils from "non-herniated" rectus sheath have uniform diameters and 640 Å periodicity with seven or more intraperiodic bands resembling typical native collagen fibrils, while the fibrils from fascia obtained from patients with direct herniation show considerable variation in diameter. These variations are often found in the same individual fibers with a range from 300 Å to 3000 Å.


Author(s):  
J.A. Nordquist ◽  
K. Chrysant ◽  
A.K. Mandal

By electron microscopy elastic tissue appear electrolucent in osmium fixed unstained grids as well as grids stained with uranyl acetate and lead citrate (UA + LC). Albert and Fleischer have studied aorta of mice with metalloporphyrins imparting conspicuous electron density to the elastic tissue. We are reporting here a modified method of electron microscopic (EM) study of the elastic tissue using metalloporhyrin, silver tetraphenyl porphyrin sulfonate (STPPS).We have studied the renal arterioles of rats and human in normal and diseased states. Elastic tissue of the aorta from young normal rat served as control for this study. Renal and aortic tissues were fixed in 4 percent glutaraldehyde, post fixed in 1 percent osmium tetroxide and embedded in spurr (blocks). From the blocks of renal tissue, 0.5 μ sections were cut, stained with methylene blue and azure II and studied by light microscopy.


Author(s):  
V. R. Mumaw ◽  
B. L. Munger

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.


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