Correlation of Ultrastructural, Morphometric and Biochemical Changes in Rats after Treatment with the Hypolipidemic Drugs Clofibrate and Probucol

1981 ◽  
Vol 39 ◽  
pp. 588-589
Author(s):  
G. E. Visscher ◽  
R. L. Robison ◽  
R. G. Engstrom
Keyword(s):  
Osmium Tetroxide ◽  
Structural Changes ◽  
Uranyl Acetate ◽  
Male Rats ◽  
Ethanol Dehydration ◽  
Sprague Dawley ◽  
Thin Sections ◽  
Biochemical Effects ◽  
Hypolipidemic Agents ◽  
Hypolipidemic Drugs

It was our goal to evaluate the reliability and reproducibility of semi-automated morphometric techniques in the analysis of structural changes observed during drug safety assessment. Studies are presented to correlate the ultrastructural, morphometric and biochemical effects that the two hypolipidemic agents, clofibrate and probucol, produce in rats.Charles River CD Sprague-Dawley derived male rats (200-225 g body wt.) were used for the three studies performed. In studies I and II, clofibrate was administered as a dietary admixture to approximate a dosage of 300 mg/kg/day for six days. In study III, clofibrate and probucol were given as dietary admixtures to approximate dosages of 300 and 250 mg/kg/day respectively for fourteen days. Processing of hepatic specimens for electron microscopy included fixation in 1.3% sym-collidine buffered osmium tetroxide, ethanol dehydration and Epon embedment. Thin sections (600Å) were stained with uranyl acetate and lead citrate. Survey and photography was performed in the manner according to Weibel. Final prints (14,400x) were analyzed with a Zeiss MOP for morphometric quantitation.

Hepatic Lipid Accumulation and Intracellular Cholesterol Crystal Formation in Hypercholesterolemic Diabetic Rats

1981 ◽  
Vol 39 ◽  
pp. 584-585
Author(s):  
Dean A. Handley ◽  
Shu Chien ◽  
Cynthia M. Arbeeny
Keyword(s):  
Cholic Acid ◽  
Uranyl Acetate ◽  
Diabetic Rats ◽  
Water Soluble ◽  
Male Rats ◽  
Crystal Formation ◽  
Sprague Dawley ◽  
Thin Sections ◽  
Hepatic Lipid Accumulation ◽  
Cellular Events

Hypercholesterolemia, induced by feeding rats a diet of cholesterol and cholic acid, can be markedly accentuated by diabetes, resulting in serum cholesterol levels greater than 1000 mg/dl and extensive accumulate of cholesterol in the liver. To examine the cellular events of hepatic cholesterol accumulation in cholesterol-fed diabetic rats, we used a fixative containing digitonin and water-soluble embedment to preserve native lipid structures.Sprague Dawley male rats were made diabetic by intravenous injection of streptozotocin (45 mg/kg body weight) and fed a powdered diet containing 2% cholesterol, 10% lard and 1% cholic acid (CLC) for 3 wk. Non-diabetic rats were also fed CLC diet. Cholesterol, glucose and insulin levels were monitored for both groups. Livers were fixed by perfusion with 3% glutaraldehyde (30°C) and 0.2% digitonin in cacodylate buffer (1 hr), then by 1% OsO4 in buffer for 2 hr. Staining was in 2% uranyl acetate and embedding in water soluble urea glutaraldehyde. Thin sections were stained with lead citrate and viewed in a Zeiss EM-9S.

Fine Structural Changes in the Adenohypophyses of Rats Following Destruction of the Pituitary Stalk

1977 ◽  
Vol 35 ◽  
pp. 496-497
Author(s):  
K. Kovacs ◽  
E. Horvath ◽  
J. M. Bilbao ◽  
F. A. Laszlo ◽  
I. Domokos
Keyword(s):  
Structural Changes ◽  
Tissue Injury ◽  
Anterior Lobe ◽  
Uranyl Acetate ◽  
Male Rats ◽  
Pituitary Stalk ◽  
Sequence Of Events ◽  
Pituitary Glands ◽  
Electrolytic Lesions ◽  
Ultrathin Sections

Electrolytic lesions of the pituitary stalk in rats interrupt adenohypophysial blood flow and result in massive infarction of the anterior lobe. In order to obtain a deeper insight into the morphogenesis of tissue injury and to reveal the sequence of events, a fine structural investigation was undertaken on adenohypophyses of rats at various intervals following destruction of the pituitary stalk.The pituitary stalk was destroyed electrolytically, with a Horsley-Clarke apparatus on 27 male rats of the R-Amsterdam strain, weighing 180-200 g. Thirty minutes, 1,2,4,6 and 24 hours after surgery the animals were perfused with a glutaraldehyde-formalin solution. The skulls were then opened and the pituitary glands removed. The anterior lobes were fixed in glutaraldehyde-formalin solution, postfixed in osmium tetroxide and embedded in Durcupan. Ultrathin sections were stained with uranyl acetate and lead citrate and investigated with a Philips 300 electron microscope.

Membranous alterations in the cytoplasm and nucleus in a primitive neuroectodermal tumor of the brain

1978 ◽  
Vol 36 (2) ◽  
pp. 628-629
Author(s):  
C. N. Sun ◽  
C. Araoz ◽  
H. J. White
Keyword(s):  
Nuclear Envelope ◽  
Tumor Cells ◽  
Osmium Tetroxide ◽  
Primitive Neuroectodermal Tumor ◽  
Uranyl Acetate ◽  
Neuroectodermal Tumor ◽  
Annulate Lamellae ◽  
Lead Citrate ◽  
Thin Sections ◽  
The Brain

The ultrastructure of a cerebral primitive neuroectodermal tumor has been reported previously. In the present case, we will present some unusual previously unreported membranous structures and alterations in the cytoplasm and nucleus of the tumor cells.Specimens were cut into small pieces about 1 mm3 and immediately fixed in 4% glutaraldehyde in phosphate buffer for two hours, then post-fixed in 1% buffered osmium tetroxide for one hour. After dehydration, tissues were embedded in Epon 812. Thin sections were stained with uranyl acetate and lead citrate.In the cytoplasm of the tumor cells, we found paired cisternae (Fig. 1) and annulate lamellae (Fig. 2) noting that the annulate lamellae were sometimes associated with the outer nuclear envelope (Fig. 3). These membranous structures have been reported in other tumor cells. In our case, mitochondrial to nuclear envelope fusions were often noted (Fig. 4). Although this phenomenon was reported in an oncocytoma, their frequency in the present study is quite striking.

Ultrastructure of myoepithelial cell in parotid gland

1988 ◽  
Vol 46 ◽  
pp. 342-343
Author(s):  
C. N. Sun
Keyword(s):  
Parotid Gland ◽  
Osmium Tetroxide ◽  
Individual Cell ◽  
Branching Processes ◽  
Muscle Cells ◽  
Myoepithelial Cells ◽  
Uranyl Acetate ◽  
Thin Sections ◽  
Gland Carcinoma ◽  
Parotid Gland Carcinoma

Myoepithelial cells have been observed in the prostate, harderian, apocrine, exocrine sweat and mammary glands. Such cells and their numerous branching processes form basket-like structures around the glandular acini. Their shapes are quite different from structures seen either in spindleshaped smooth muscle cells or skeletal muscle cells. These myoepithelial cells lie on the epithelial side of the basement membrane in the glands. This presentation describes the ultrastructure of such myoepithelial cells which have been found also in the parotid gland carcinoma from a 45-year old patient.Specimens were cut into small pieces about 1 mm3 and immediately fixed in 4 percent glutaraldehyde in phosphate buffer for two hours, then post-fixed in 1 percent buffered osmium tetroxide for 1 hour. After dehydration, tissues were embedded in Epon 812. Thin sections were stained with uranyl acetate and lead citrate. Ultrastructurally, the pattern of each individual cell showed wide variations.

Electron Microscopic Examination of a Transplantable Rat Mammary Carcinoma

1968 ◽  
Vol 26 ◽  
pp. 20-21
Author(s):  
S. Shirahama ◽  
G. C. Engle ◽  
R. M. Dutcher
Keyword(s):  
Electron Microscope ◽  
Osmium Tetroxide ◽  
Electron Microscopic Examination ◽  
Undifferentiated Carcinoma ◽  
Uranyl Acetate ◽  
Sprague Dawley ◽  
Electron Microscopic ◽  
North West ◽  
X Irradiation ◽  
Tissue Specimens

A transplantable carcinoma was established in North West Sprague Dawley (NWSD) rats by use of X-irradiation by Engle and Spencer. The tumor was passaged through 63 generations over a period of 32 months. The original tumor, an adenocarcinoma, changed into an undifferentiated carcinoma following the 19th transplant. The tumor grew well in NWSD rats of either sex at various ages. It was invariably fatal, causing death of the host within 15 to 35 days following transplantation.Tumor, thymus, spleen, and plasma from 7 rats receiving transplants of tumor at 3 to 9 weeks of age were examined with an electron microscope at intervals of 8, 15, 22 and 30 days after transplantation. Four normal control rats of the same age were also examined. The tissues were fixed in glutaraldehyde, postfixed in osmium tetroxide and embedded in Epon. The plasma was separated from heparanized blood and processed as previously described for the tissue specimens. Sections were stained with uranyl acetate followed by lead citrate and examined with an RCA EMU-3G electron microscope.

Observations of thick and thin sections in a field-emission SEM

1994 ◽  
Vol 52 ◽  
pp. 1018-1019
Author(s):  
W. P. Wergin ◽  
S. Roy ◽  
E. F. Erbe ◽  
C. A. Murphy ◽  
C. D. Pooley
Keyword(s):  
Electron Microscope ◽  
Field Emission ◽  
Room Temperature ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Chemical Fixation ◽  
Thin Sections ◽  
Transmission Electron ◽  
Conventional Transmission Electron Microscope ◽  
Scanning Electron

Larvae of the nematode, Steinernema carpocapsae Weiser strain All, were cryofixed and freezesubstituted for 3 days in acetone containing 2% osmium tetroxide according to established procedures. Following chemical fixation, the nematodes were brought to room temperature, embedded in Spurr's medium and sectioned for observation with a Hitachi S-4100 field emission scanning electron microscope that was equipped with an Oxford CT 1500 Cryotrans System. Thin sections, about 80 nm thick, similar to those generally used in conventional transmission electron microscope (TEM) studies were mounted on copper grids and stained with uranyl acetate for 30 min and lead citrate for 5 min. Sections about 2 μm thick were also mounted and stained in a similar fashion. The grids were mounted on an Oxford grid holder, inserted into the microscope and onto a cryostage that was operated at ambient temperature. Thick and thin sections of the larvae were evaluated and photographed in the SEM at different accelerating voltages. Figs. 4 and 5 have undergone contrast conversion so that the images would resemble transmitted electron micrographs obtained with a TEM.

An Electron Microscopic Investigation of the Pathogenesis of Hemorrhage Induced by Rattlesnake Venom

1974 ◽  
Vol 32 ◽  
pp. 56-57
Author(s):  
Charlotte L. Ownby ◽  
Robert A. Kainer ◽  
Anthony T. Tu
Keyword(s):  
Phosphate Buffer ◽  
Osmium Tetroxide ◽  
Microscopic Investigation ◽  
Uranyl Acetate ◽  
Electron Microscopic Investigation ◽  
Lead Citrate ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Rattlesnake Venom ◽  
Diamondback Rattlesnake

One of the significant changes induced by the injection of rattlesnake (Crotalidae) venom is hemorrhage. Since crotaline antivenin does not prevent such local tissue damage, a more effective treatment of snakebite is needed. To aid in the development of such a treatment the pathogenesis of venom-induced hemorrhae was investigated.Swiss-Webster white mice were injected intramuscularly with Western diamondback rattlesnake (Crotalus atrox) venom. Two minutes after the injection, muscle tissue was obtained by bioosy from the thigh and fixed in 6% glutaraldehyde in Milloniq's phosphate buffer (DH 7.4, 2 hrs., 4°C). After post-fixation in 2% osmium tetroxide in Milloniq's phosphate buffer (pH 7.4, 1hr., 4°C) the tissue was dehydrated routinely in ethanol and embedded in Epon 812. The thin sections were stained with uranyl acetate in methanol and lead citrate then observed with either a Zeiss EM 9A or an Hitachi HS-8 electron microscope.

Ultrastructure of Granules Passing Through Frog Oocyte Nuclear Pores

1969 ◽  
Vol 27 ◽  
pp. 288-289
Author(s):  
Madison B. Cole
Keyword(s):  
Osmium Tetroxide ◽  
Rana Pipiens ◽  
Uranyl Acetate ◽  
Nuclear Pores ◽  
Thin Sections ◽  
Close Proximity ◽  
Perinuclear Cytoplasm ◽  
Frog Oocyte ◽  
Pass Through ◽  
Number Of Authors

Ovaries of Rana pipiens larvae were fixed in glutaraldehyde, post-fixed in osmium tetroxide, and embedded in epon. Thin sections of oocytes were routinely stained using methanolic uranyl acetate followed by lead citrate.The granule, designated M1, found in the primary oocytes is considered to pass through the nuclear pores by virtue of 1) its location in the nucleoplasm in close proximity to the nuclear envelope, 2) its contact with the nuclear pores, 3) its presence in aggregates in the perinuclear cytoplasm and A) its similarity to granules referred to by a number of authors (1-4) as material passing through nuclear pores.

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.

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