Electron Microscopic Examination of a Transplantable Rat Mammary Carcinoma

Author(s):  
S. Shirahama ◽  
G. C. Engle ◽  
R. M. Dutcher

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.

Author(s):  
Iracema M. Baccarini

The embryology of granulosa and theca cells is not understood thoroughly. Electron microscopic studies in the past have been concerned mainly with mature granulosa cells and less with their development.Material and Methods. Rat fetuses were removed surgically under ether anesthesia at 16-17, 17-18 and 18-19 days of gestation. Their abdominal cavities were opened, and the fetuses were placed immediately into 3% glutaraldehyde (pH 7.2) for 3 hours. During this time, the fetal ovaries were dissected under a microscope. The tissue was washed in phosphatebuffer for 24 hours, post-fixed in 1% phosphate buffered osmium tetroxide for 1-2 hours at 4°C, and embedded in Durcupan ACM (Fluka). Sections were double stained with uranyl acetate and lead citrate, and viewed in an RCA-EMU-3D electron microscope.


Author(s):  
W. G. Banfield ◽  
G. Kasnic ◽  
J. H. Blackwell

An ultrastructural study of the intestinal epithelium of mice infected with the agent of epizootic diarrhea of infant mice (EDIM virus) was first performed by Adams and Kraft. We have extended their observations and have found developmental forms of the virus and associated structures not reported by them.Three-day-old NLM strain mice were infected with EDIM virus and killed 48 to 168 hours later. Specimens of bowel were fixed in glutaraldehyde, post fixed in osmium tetroxide and embedded in epon. Sections were stained with uranyl magnesium acetate followed by lead citrate and examined in an updated RCA EMU-3F electron microscope.The cells containing virus particles (infected) are at the tips of the villi and occur throughout the intestine from duodenum through colon. All developmental forms of the virus are present from 48 to 168 hours after infection. Figure 1 is of cells without virus particles and figure 2 is of an infected cell. The nucleus and cytoplasm of the infected cells appear clearer than the cells without virus particles.


Author(s):  
Iracema M. Baccarini

Some morphological nuclear features (invaginations) in normal and abnormal cells have been described in several electron microscopic studies. They have been referred to by others as blebs, loops, pockets, sheets, bodies, nuclear inclusions and cytoplasmic invaginations. Identical appearing structures were found in cells of the uterine cervical epithelium, in trophoblasts of blastocysts and in trophoblasts of rat placenta.Methods. Uterine cervix (normal rats), rat placenta (9-10 days gestation) and blastocyst were placed in 3% glutarahdehyde for 3 hours. The tissue was washed in phosphate buffer for 24 hours, postfixed in 1%. buffered osmium tetroxide for 1-2 hours and embedded in epon araldite. Sections were double stained with uranyl acetate and lead citrate and viewed in E. M. Siemens 200.Observations. Nuclear invaginations were found in basal, parabasal and mucous cells of the cervix epithelium, in trophoblasts of blastocyst and in trophoblasts of placenta. An oval, round or elongated invagination contained heterogenously cytoplasm surrounded by a double intact membrane; usually several invaginations were found in the same nucleus.


Author(s):  
R.C. Caughey ◽  
U.P. Kalyan-Raman

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.


Author(s):  
W. P. Wergin ◽  
S. Roy ◽  
E. F. Erbe ◽  
C. A. Murphy ◽  
C. D. Pooley

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.


Author(s):  
Caroline A. Miller ◽  
David H. Nichols ◽  
Richard F. Murphy

Gastrin is a small peptide capable of both stimulating gastric acid secretion and acting as an enteric growth factor. Known functions of eosinophils in the rat stomach are related to immunological defense. Here we demonstrate the binding of biotinylated gastrin to rat stomach eosinophils in the electron microscope. Small pieces of stomach were fixed by immersion in 4% paraformaldehyde/0.1% glutaraldehyde in 0.1 M phosphate buffer, pH 7.4 for 1 hour. The tissue was then cryoprotected in 30% sucrose/0.1 M phosphate buffer, transferred to Tissue Tek OCT compound and frozen in isopentane cooled with liquid nitrogen. Transverse cryostat sections were cut at 25 μm, thawed in PBS and free floating sections exposed to 10−5 M biotinylated 1-17 gastrin (human sequence; Peninsula Labs) for 1 hour. Controls omitted the biotinylated gastrin from this step. Sections were then rinsed 3X in PBS and exposed to either:1).a 1:50 dilution of 10 nm Extravidin colloidal gold (Sigma) for 2 hours, or2).an avidin-biotin-alkaline phosphatase complex (ABC-AP;Vector) for 1 hour. A substrate solution containing cerium chloride was used to generate an electron dense reaction product.Sections from both procedures were postfixed in 1% OsO4 in 0.1 M phosphate buffer, rinsed and dehydrated. These were then flat embedded in EMbed 812 between two microscope slides coated with Liquid Release (both from Electron Microscopy Sciences).Polymerized sections were adhered to resin blocks using super glue, cut at 70-90 nm, stained with uranyl acetate/lead citrate and observed in a Philips CM-10 electron microscope.


2016 ◽  
Vol 90 (4) ◽  
pp. 741-762 ◽  
Author(s):  
Brian L. Hoffman ◽  
Scott A. Hageman ◽  
Gregory D. Claycomb

AbstractAlthough Ptychodus teeth are well known in Late Cretaceous marine deposits in North America and Europe and a few specimens with jaw elements have been discovered, the taxonomic position of the shark genus Ptychodus is enigmatic due to the lack of preservation of diagnostic material other than teeth. These sharks possessed a pavement dentition suited to a diet of hard-shelled macroinvertebrates (durophagy), leading several studies to variously describe Ptychodus as a batoid, a hybodont shark, or a selachimorph. Members of the Selachimorpha share one dental synapomorphy, a triple-layered enameloid (TLE) consisting of an outer shiny-layered enameloid (SLE) of randomly oriented hydroxyapatite crystallites, a middle layer of parallel-bundled enameloid (PBE), and an inner layer of tangled-bundled enameloid (TBE). Batoids and hybodonts both have teeth with single crystallite enameloid (SCE). We examined teeth from Ptychodus collected from the Lincoln Limestone of the Greenhorn Formation of Barton County, Kansas, and compared their enameloid ultrastructure with that of a Carboniferous hybodontiform and the Cretaceous lamniform shark Squalicorax curvatus Williston, 1900. Scanning electron microscopic examination of Ptychodus shows that crystallite bundling in the form of a TLE is evident in these teeth. The PBE is most apparent at transverse enameloid ridges of Ptychodus teeth. Columns of dentine penetrate into the tooth enameloid, and the crystallites near the dentine are randomly oriented. These observations bolster the argument that Ptychodus is a genus of highly specialized selachimorph shark, rather than a hybodont or batoid.


1973 ◽  
Vol 40 (1) ◽  
pp. 113-117 ◽  
Author(s):  
M. H. Abd El-Salam ◽  
Safinaz El-Shibiny

SummaryA technique is described for preparing ultrathin sections from cheese for electron-microscopic examination. The internal structure of fresh Domiati cheese was found to be composed of a framework of large, spherical casein aggregates held by bridges and enclosing fat.After pickling, the casein aggregates were partly disintegrated into small spherical particles forming a loose structure.


Author(s):  
Charlotte L. Ownby ◽  
Robert A. Kainer ◽  
Anthony T. Tu

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.


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.


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