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.

scholarly journals Ultrastructural visualization of complex carbohydrates in epiphyseal cartilage with the tannic acid-metal salt methods.

1983 ◽  
Vol 31 (6) ◽  
pp. 783-790 ◽  
Author(s):  
M Takagi ◽  
R T Parmley ◽  
F R Denys ◽  
M Kageyama
Keyword(s):  
Tannic Acid ◽  
Metal Salt ◽  
Secretory Granules ◽  
Uranyl Acetate ◽  
Collagen Fibrils ◽  
Epiphyseal Cartilage ◽  
Thin Sections ◽  
Background Staining ◽  
Complex Carbohydrates ◽  
Cytoplasmic Glycogen

The present study has ultrastructurally applied the tannic acid-ferric chloride (TA-Fe) and the TA-uranyl acetate (TA-UA) methods to thin sections of glutaraldehyde-fixed, unosmicated embedded epiphyseal cartilage from rat tibiae to demonstrate complex carbohydrates. The strongest TA-Fe and TA-UA staining was observed after fixation of the specimens in glutaraldehyde containing TA. TA-Fe (pH 1.5) strongly stained matrix granules presumed to be proteoglycan monomers and chondrocyte secretory granules at various maturational stages but did not stain collagen fibrils and glycogen. TA-UA (pH 4.2) strongly stained matrix granules, intracellular glycogen, and chondrocyte secretory granules, and moderately stained collagen fibrils in the cartilage matrix. Ribosomes and nuclei were not stained above background staining with UA alone. In alpha-amylase-digested specimens, all TA-UA-reactive cytoplasmic glycogen was selectively removed. Testicular hyaluronidase digestion of specimens selectively removed TA-UA staining in matrix granules and all TA-Fe staining. When the pH of the UA solution was reduced to 1.5, TA-UA staining of glycogen and collagen was markedly decreased or absent, whereas staining of anionic sites was unaltered and significantly greater than with UA staining alone. Thus the TA-metal salt methods are pH dependent and allow differential intracellular and extracellular localization of complex carbohydrates in cartilage tissues at the electron microscope 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.

scholarly journals Ultrastructural visualization of complex carbohydrates in odontoblasts, predentin, and dentin matrix by the tannic acid-uranyl acetate method.

1985 ◽  
Vol 27 (4) ◽  
pp. 211-217 ◽  
Author(s):  
Masato KAGEYAMA ◽  
Hidehito KADO ◽  
Noboru KAJIYAMA ◽  
Kunihito KADO ◽  
Ken NAGAI ◽  
...  
Keyword(s):  
Tannic Acid ◽  
Uranyl Acetate ◽  
Dentin Matrix ◽  
Complex Carbohydrates

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.

scholarly journals Electron microscopic localization of chromogranin A in osmium-fixed neuroendocrine cells with a protein A-gold technique.

1987 ◽  
Vol 35 (7) ◽  
pp. 795-801 ◽  
Author(s):  
S A Hearn
Keyword(s):  
Neuroendocrine Tumors ◽  
Chromogranin A ◽  
Osmium Tetroxide ◽  
Secretory Granules ◽  
Protein A ◽  
Neuroendocrine Cells ◽  
Neurosecretory Granules ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Carotid Body Tumors

An antibody (LK2H10) to chromogranin A has been recommended for use in ultrastructural identification of neuroendocrine secretory granules. Previous studies have demonstrated immunoreactive chromogranin A in specimens prepared for electron microscopy by glutaraldehyde fixation only. In this study, the effect of specimen post-fixation by osmium tetroxide on post-embedding localization of chromogranin A was evaluated. Human tissues from benign endocrine glands, neuroendocrine tumors, and non-neuroendocrine tumors were post-fixed in osmium, embedded in epoxy resin, and the sample thin sections immunolabeled using a protein A-gold technique. Chromogranin A-positive neurosecretory granules were detected in pancreatic islets, adrenal medulla, stomach, ileum, anterior pituitary, and parathyroid. Mid-gut carcinoids, bronchial carcinoids, pheochromocytomas, paragangliomas, carotid body tumors, and thyroid medullary carcinomas contained immunoreactive granules. Cytoplasmic granules in non-neuroendocrine tumors did not react for chromogranin A. Tissues post-fixed in osmium tetroxide had optimally preserved ultrastructural features, and use of this fixative is compatible with postembedding localization of chromogranin A in neurosecretory granules.

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.

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).

Estrogen and progesterone receptors in carcinoma of the breast (A cytochemical, immunofluorescence and electron microscopic study)

1986 ◽  
Vol 44 ◽  
pp. 116-117
Author(s):  
Robert L. Elliott ◽  
Gregory O. Harrison ◽  
Anne Coble-Landry ◽  
Lindsay Bullock-Ledford ◽  
Anne T. McRea ◽  
...  
Keyword(s):  
Ductal Carcinoma ◽  
Progesterone Receptors ◽  
Uranyl Acetate ◽  
Staining Procedure ◽  
Electron Microscopic ◽  
Carcinoma Of The Breast ◽  
Thin Sections ◽  
Infiltrating Ductal Carcinoma ◽  
Estrogen And Progesterone Receptor ◽  
Dual Staining

Tissue from 105 patients with infiltrating ductal carcinoma of the breast was assayed for the estrogen and progesterone receptor by the cytochemical dextran coated charcoal technique and by immunofluorescence with a direct dual staining procedure with Fluorocep, and also examined electron microscopically.Tissue was fixed in Zambonis; post-fixed in 1% OsO4; embedded in a Med Cast Resin, and thin sections were cut on a ultramicrotone. They were stained with uranyl acetate and lead citrate and examined in a Hitachi H-600 electron microscope.

The response of cell walls of Bacillus subtilis to metals and to electron-microscopic stains

1978 ◽  
Vol 24 (2) ◽  
pp. 89-104 ◽  
Author(s):  
T. J. Beveridge
Keyword(s):  
Bacillus Subtilis ◽  
Electron Scattering ◽  
Cell Walls ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Ruthenium Red ◽  
Transition Elements ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Wall Ultrastructure

Purified cell walls of Bacillus subtilis were subjected to solutions of 40 independent metals and the metal uptake, the electron-scattering power of thin sections, and the type of staining response evaluated. This was repeated for six typical electron-microscopic stains (uranyl acetate, uranyl magnesium acetate, osmium tetroxide, Os-meth, osmium-dimethylethylenediamine, and ruthenium red) and one new staining reagent (a potassium platinum chloride – dimethylsulfoxide complex) whose specificity is for amine functions. The reaction of select metals can be specific in terms of both uptake and staining response. Of the metals studied most transition elements had a high affinity for the wall fabric and some (i.e., Sc III, most lanthanides, U IV, Zr IV, Hf IV, Fe III, Pd II, Ru III, and In III) may be suitable as contrasting agents for electron microscopy. Furthermore, when the thickness of metal-reacted walls was compared to freeze-each and ultracryotomy data, statistical-dimensional differences were commonly seen, which indicates that wall ultrastructure can be profoundly affected by the type of metal and (or) staining reagent.

Electron microscopy of the microangiopathy and immunohistochemistry of the inflammatory infiltrate in eosinophilia-myalgia syndrome

1991 ◽  
Vol 49 ◽  
pp. 24-25
Author(s):  
Stephen A. Smith
Keyword(s):  
Electron Microscopy ◽  
B Cell ◽  
Inflammatory Infiltrate ◽  
Uranyl Acetate ◽  
T Helper ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Microscopic Appearance ◽  
Muscle Fascia ◽  
Eosinophilia Myalgia Syndrome

The ingestion of L-tryptophan (LT) was linked with the eosinophilia-myalgia syndrome (EMS) in the fall of 1989. The pathology is a microangiopathy consisting of endothelial cell dysfunction and invasion of blood vessels by lymphocytes. Perimysial, fascial, and dermal perivascular lymphocytes and eosinophils are present. A contaminant in the manufacturing process of LT has been identified as the di-L-tryptophan aminal of acetaldehyde. This report describes the electron microscopic appearance of the microangiopathy and the phenotyping of the inflammatory infiltrate by immunohistochemistry.Muscle, fascia, and skin biopsies from 21 affected individuals were rapidly frozen for light microscopy, histochemistry, and immunohistochemistry. For electron microscopy tissues were preserved in 4% formaldehyde-1% glutaraldehyde with post-fixation in osmium tetroxide. Thin sections were stained with uranyl acetate and lead citrate and examined in a JEOL 100CX electron microscope. Eight anlisera were used for the immunohistochemistry to identify lymphocyte subsets and macrophages. The markers were CD-2 (pan T), CD-3 (pan T), CD-4 (T helper), CD-5 (T helper), CD-8 (T suppressor), CD-19 (B cell), CD- 22 (B cell), and CD-68 (macrophage). Three high power fields were examined from the dermis, fascia, and muscle from eight biopsies counting the immunoperoxidase labelled cells for each marker.

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