scholarly journals ELECTRON MICROSCOPIC RADIOAUTOGRAPHY OF THIN SECTIONS: THE GOLGI ZONE AS A SITE OF PROTEIN CONCENTRATION IN PANCREATIC ACINAR CELLS

1961 ◽  
Vol 10 (1) ◽  
pp. 37-45 ◽  
Author(s):  
Lucien G. Caro
Keyword(s):  
Guinea Pig ◽  
Golgi Complex ◽  
Optical Resolution ◽  
Nuclear Research ◽  
Uranyl Acetate ◽  
Thin Layers ◽  
Pancreatic Acinar Cells ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Electron Microscopic Radioautography

Electron microscopic radioautographs of guinea pig pancreatic exocrine cells were obtained by covering thin sections (∼ 600 A) of OsO4-fixed, methacrylate-embedded tissue with thin layers of Ilford K-5 nuclear research emulsion. After an exposure of 13 days at 4°C., the preparations were photographically processed, stained with uranyl acetate, and examined in an electron microscope. The label used was leucine-H3 injected intravenously 20 minutes before collection of the specimens. Conventional radioautographs of thicker sections (0.4 micron) were also examined in a phase contrast microscope. The advantages obtained from electron microscopic radioautography are: the higher radioautographic resolution (of the order of 0.3 micron) due to the thinness of the emulsion and the specimen, and a high optical resolution permitting a clear identification of the labeled structure. In the guinea pig pancreas this technique demonstrated that, at the time studied, newly synthesized proteins were concentrated in the structures of the Golgi complex and especially in large vacuoles partially filled with a dense material. The vacuoles are probably a precursor to the secretion granules (zymogen granules) in which the label becomes segregated at a later time. These observations demonstrate directly the role of the Golgi complex in the secretion process. They also illustrate the possibilities of this method for radioautography at the intracellular level.

An Electron Microscopic Study of the Acute Effects of Hypothalamic Releasing Factors on the Anterior Pituitary Gland

1968 ◽  
Vol 26 ◽  
pp. 232-233
Author(s):  
P.W. Coates ◽  
E.A. Ashby ◽  
L. Krulich ◽  
A. Dhariwal ◽  
S. McCann
Keyword(s):  
Anterior Pituitary ◽  
Microscopic Investigation ◽  
Uranyl Acetate ◽  
Electron Microscopic Investigation ◽  
Electron Microscopic ◽  
Tissue Samples ◽  
Thin Sections ◽  
Dense Membrane ◽  
Membrane Bound ◽  
Releasing Factors

The morphologic effects on somatotrophs of crude sheep hypothalamic extract prepared from stalk-median eminence were studied by electron microscopy in conjunction with concurrently run bioassays performed on the same tissue samples taken from young adult male Sherman rats.Groups were divided into uninjected controls and injected experimentals sacrificed at 5', 15', and 30' after injection. Half of each anterior pituitary was prepared for electron microscopic investigation, the other half for bioassay. Fixation using collidine buffered osmium tetroxide was followed by dehydration and embedment in Maraglas. Uranyl acetate and lead citrate were used as stains. Thin sections were examined in a Philips EM 200.Somatotrophs from uninjected controls appeared as described in the literature (Fig. 1). In addition to other components, these cells contained moderate numbers of spherical, electron-dense, membrane-bound granules approximately 350 millicrons in diameter.

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.

Electron microscopic evidence for a double hair-like nap appearing at low frequency on Bacillus anthracis Sterne spores

1969 ◽  
Vol 15 (10) ◽  
pp. 1247-1248 ◽  
Author(s):  
M. J. Kramer ◽  
Ivan L. Roth
Keyword(s):  
Bacillus Anthracis ◽  
Microscopic Examination ◽  
Low Frequency ◽  
Electron Microscopic Examination ◽  
Uranyl Acetate ◽  
Spore Coat ◽  
Lead Citrate ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Very Low Frequency

Electron microscopic examination of thin sections of Bacillus anthracis Sterne spores triply poststained with KMnO4, uranyl acetate, and lead citrate has indicated an unusual morphological variant. These spores are seen at very low frequency and have, in addition to the hair-like nap normally associated with the exosporium a second hairy layer which appears to originate in the spore coat complex.

Comparison of Cholesterol Extraction from Tissues During Processing for Electron Microscopic Radioautography

1968 ◽  
Vol 26 ◽  
pp. 92-93
Author(s):  
E. Tessa Hedley-Whyte ◽  
Betty G. Uzman
Keyword(s):  
Electron Microscope ◽  
Lipid Metabolism ◽  
Propylene Oxide ◽  
Mouse Liver ◽  
Uranyl Acetate ◽  
Electron Microscopic ◽  
Electron Microscopic Radioautography

Radioautographic studies of lipid metabolism with the electron microscope have been limited by the large losses of lipid which occur during the usual dehydration and infiltration procedures. In this study different methods of processing tissue for electron microscopic radioautography have been compared with respect to the extraction of cholesterol-1, 2-3H from labeled mouse liver and nerve.In all methods primary fixation had been in 10% formalin for several months followed by washing overnight. Post-fixation for one hour in Dalton's chrome osmium and staining in 1% uranyl acetate in 10% formalin resulted in small but constant losses. Four methods were assessed: 1) graded acetone dehydration with propylene oxide and epon-araldite mixture infiltration, 2) exposure to 0.5% digitonin in 50% ethanol for 1 hour before post-fixing in osmium followed by procedure 1, 3) Durcupan embedding, and 4) limited dehydration followed by embedding in an epon:araldite mixture (modified Idelman).

scholarly journals Electron Microscopic Observations of the Carotid Body of the Cat

1959 ◽  
Vol 6 (2) ◽  
pp. 253-262 ◽  
Author(s):  
Leonard L. Ross
Keyword(s):  
Golgi Complex ◽  
Carotid Body ◽  
Nerve Fibers ◽  
Osmic Acid ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Sustentacular Cells ◽  
Carotid Bodies ◽  
Unmyelinated Nerve ◽  
Chemoreceptor Cells

Carotid bodies were removed from cats, fixed in buffered 1 per cent osmic acid, embedded in deaerated, nitrogenated methacrylate, and cut into thin sections for electron microscopic study. The carotid body is seen to be composed of islands of chemoreceptor and sustentacular cells surrounded by wide irregular sinusoids. These cells are separated from the sinusoids by relatively broad interstitial spaces which are filled with collagen, fibroblasts, and many unmyelinated nerve fibers with their Schwann cell sheaths. The chemoreceptor cells are surrounded by the flattened, multiprocessed sustentacular cells which serve to convey the axons from an interstitial to a pericellular location. These sustentacular cells are assumed to be lemmoblastic in origin. Relatively few axons are seen to abut on the chemoreceptor cells. The cytoplasm of the chemoreceptor cell is characterized by numerous small mitochondria, units of granular endoplasmic reticulum, a small Golgi complex, and a variety of vesicles. There are many small vesicles diffusely scattered throughout the cytoplasm. In addition, there is a small number of dark-cored vesicles of the type which has been previously described in the adrenal medulla. These are usually associated with the Golgi complex. These findings are discussed in relation to the concepts of the origin of the chemoreceptor cell and the nature of the synapse.

scholarly journals Appendix: Radioautographic localization of labelled proteins after incubation of guinea-pig islets of Langerhans with [3H]tryptophan

1974 ◽  
Vol 140 (1) ◽  
pp. 22-23 ◽  
Author(s):  
S. L. Howell ◽  
C. Hellerström ◽  
M. Whitfield
Keyword(s):  
B Cells ◽  
Guinea Pig ◽  
B Cell ◽  
Islets Of Langerhans ◽  
Electron Microscopic ◽  
Storage Granules ◽  
Silver Grains ◽  
Electron Microscopic Radioautography ◽  
Significant Concentration

An analysis was made by electron-microscopic radioautography of the distribution of silver grains over storage granules of A2 and B cells after incubation of isolated guinea-pig islets of Langerhans for 17h in the presence of [3H]tryptophan. A significant concentration of labelled proteins was present in the A2 cell, but not in the B-cell granules.

scholarly journals Tannic acid-metal salt sequences for light and electron microscopic localization of complex carbohydrates.

1978 ◽  
Vol 26 (1) ◽  
pp. 55-61 ◽  
Author(s):  
P L Sannes ◽  
T Katsuyama ◽  
S S Spicer
Keyword(s):  
Tannic Acid ◽  
Ferric Chloride ◽  
Metal Salt ◽  
Secretory Granules ◽  
Uranyl Acetate ◽  
Gold Chloride ◽  
High Concentration ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Complex Carbohydrates

Use of tannic acid (TA), in sequence with ferric chloride, uranyl acetate or gold chloride resulted in staining of selective but sometimes different sites in paraffin sections. TA-uranyl acetate of TA-ferric chloride stained sites rich in complex carbohydrates, wherease TA-gold chloride stained the collagen of various connective tissues different shades of red-purple to gray-black. Applied to epoxy-embedded thin sections of tissues fixed with glutaraldehyde and not post-osmicated, TA-uranyl acetate and TA-ferric chloride imparted density to subcellular sites known to contain a high concentration of mucosubstances, such as secretory granules and cisternae of the Golgi complex of certain cells. TA-gold chloride proved unsatisfactory for ultracytochemistry because of its tendency to form globular precipitates on thin sections. The effect of blockage procedures at the light microscopic level indicated that vicinal glycols are not required for binding of TA to tissue sites. Electrostatic forces were shown to be of minimal significance, whereas hydrogen bonding appeared to play a part in both TA-tissue and TA-metal binding mechanisms.

Electron microscopic radioautographic study of glycoprotein biosynthesis and renewal in the renal glomeruli of the rat

1978 ◽  
Vol 36 (2) ◽  
pp. 476-477
Author(s):  
A. Haddad ◽  
J-J. Lachat ◽  
R. P. Gonçalves
Keyword(s):  
Renal Cortex ◽  
Mesangial Cells ◽  
Electron Microscopic ◽  
Time Intervals ◽  
Light Microscope Level ◽  
Thin Sections ◽  
Main Site ◽  
Renal Glomeruli ◽  
Silver Grains ◽  
Electron Microscopic Radioautography

Biosynthesis, migration and renewal of glycoproteins were studied by radioautography in the renal glomeruli after administering L-3H-fucose to rats which were killed at several time intervals after injection, by perfusion of glutaraldehyde through the abdominal aorta. Small pieces of the kidney were post fixed in 0s04, dehydrated and embedded in Epon 812. Semithin and thin sections of the renal cortex were processed for light and electron microscopic radioautography, respectively. At the light microscope level (Fig. 1) it was observed that the main site of incorporation of 3H-fucose into glycoproteins was the paranuclear region of the visceral epithelial cells (podocytes). Weak paranuclear radioautographic reactions were also observed in endothelial and mesangial cells at 10 minutes after injection. At the longer time intervals these paranuclear reactions disappeared and the silver grains were mostly overlying the several components of the capillary wall. Silver grain counts showed that the peak of the silver grain density over the glomeruli occurred at 4 hours after injection; by 14 days the radioautographic reactions were almost negligible.

Ultrastructure of Oviduct Epithelium of the Porcine in Estrus

1971 ◽  
Vol 29 ◽  
pp. 512-513
Author(s):  
R. K. Nayak ◽  
D. R. Zimmerman
Keyword(s):  
Intercellular Junctions ◽  
Uranyl Acetate ◽  
Secretory Process ◽  
Osmic Acid ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Tissue Sections ◽  
Oviduct Epithelium ◽  
Microscopic Studies ◽  
Cyclic Changes

Cyclic changes of porcine oviduct epithelium studied by light microscopy were first reported by Snyder in 1923. UltrastructuraI features of the porcine oviduct epithelium have not yet been described. Electron microscopic studies of oviduct epithelium have been reported for only a few species. These reports have been recently reviewed by Nilsson and Relnius. This report describes the fine structure of, the oviduct epithelium and attempts to elucidate the mechanism of ciliogenesis and the secretory process in the porcine during estrus.Tissue sections from the fimbria and ampulla were fixed in cold 3% cacodylate buffered glutaraldehyde (pH 7.4), post-fixed in 1% osmic acid and embedded in Epon. Ultra-thin sections were stained with uranyl acetate and lead citrate and examined in a RCA 3-G electron microscope operated at 100 kv.The epithelium of the tubal mucosa consists of secretory and ciliated cells. The cells are columnar and rest on a common basement membrane, which is about 50 mμ thick. The distal or free borders of the surface epithelial cells possess few irregular microvilli. The membranes of adjacent cells show tight intercellular junctions and macula adhaerentes (Figs. 1, 2).

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