Antikeratin antibody staining on ultrathin sections of epidermal cells prepared by low-denaturation embedding

1985 ◽  
Vol 91 (2) ◽  
pp. 92-103 ◽  
Author(s):  
Kenneth D. Marenus
Keyword(s):  
Epidermal Cells ◽  
Antibody Staining ◽  
Ultrathin Sections ◽  
Antikeratin Antibody

Ferritin Labeled Antibody Staining of Thin Sections

1969 ◽  
Vol 27 ◽  
pp. 420-421
Author(s):  
J. D. McLean ◽  
S. J. Singer
Keyword(s):  
Biological Material ◽  
Water Soluble ◽  
Thin Sections ◽  
Antibody Staining ◽  
Thin Sectioning ◽  
Ultrathin Sections

The successful application of ferritin labeled antibodies (F-A) to ultrathin sections of biological material has been hampered by two main difficulties. Firstly the normally used procedures for the preparation of material for thin sectioning often result in a loss of antigenicity. Secondly the polymers employed for embedding may non-specifically absorb the F-A. Our earlier use of cross-linked polyampholytes as embedding media partially overcame these problems. However the water-soluble monomers used for this method still extract many lipids from the material.

scholarly journals ULTRASTRUCTURAL LOCALIZATION OF CONTRACTILE PROTEIN (THROMBOSTHENIN) IN HUMAN PLATELETS USING AN UNLABELED ANTIBODY-PEROXIDASE STAINING TECHNIQUE

1971 ◽  
Vol 19 (9) ◽  
pp. 540-550 ◽  
Author(s):  
FRANCOIS M. BOOYSE ◽  
DOROTHEA ZSCHOCKE ◽  
MAX E. RAFELSON ◽  
LUDWIG A. STERNBERGER
Keyword(s):  
Horseradish Peroxidase ◽  
Staining Technique ◽  
Ultrastructural Localization ◽  
Human Platelets ◽  
Contractile Protein ◽  
Intact Cells ◽  
Proteinaceous Material ◽  
Antibody Staining ◽  
Ultrathin Sections ◽  
Unlabeled Antibody

Soluble horseradish peroxidase-antihorseradish peroxidase (rabbit) complex was used to localize the actomyosin-like contractile protein, thrombosthenin, in both intact human platelets (Epon-embedded) and ultrathin sections (methacrylate-embedded). Antibody staining of ultrathin sections showed the presence of membrane-associated and cytoplasmic thrombosthenin. Antibody staining of intact cells showed that membrane-associated thrombosthenin was localized, at least in part, in the exterior, "fluffy" coat of the platelet. Even after prolonged fixation and/or incubation with the various antisera, we were unable to demonstrate antibody penetration of intact, fixed platelets. Brief treatment with Pronase removed the surface-localized proteinaceous material and completely abolished all antibody staining.

scholarly journals THE BASEMENT LAMELLA OF AMPHIBIAN SKIN

1956 ◽  
Vol 2 (4) ◽  
pp. 275-282 ◽  
Author(s):  
Paul Weiss ◽  
Wayne Ferris
Keyword(s):  
Layered Structure ◽  
Epidermal Cells ◽  
Ground Substance ◽  
Amphibian Skin ◽  
Diameter Class ◽  
Amphibian Larvae ◽  
Sequence Of Events ◽  
Epidermal Surface ◽  
Ultrathin Sections ◽  
Geometric Regularity

The basement lamella under the epidermis of amphibian larvae shows a sub-microscopic architecture of remarkable geometric regularity: It consists of about twenty layers of ground substance in which cylindrical fibers (presumably collagenous) of about 500 Angström diameter are embedded parallel to one another, but with the fiber directions alternating by 90° from layer to layer. The repair of this membrane after wounding was studied electronmicroscopically in ultrathin sections. The sequence of events is as follows: (1) Epidermal cells cover the wound exudate by migration. (2) Rather uniform fibers of small size (<200 A) appear in the space between the epidermal underside and the subjacent fibroblasts; these fibers are sparse and oriented at random. (3) Proceeding from the epidermal surface downward, a wave of organization spreads over this primitive fiber tangle, resulting in the fibers becoming (a) straightened; (b) oriented; (c) packed into the characteristic layered structure; and (d) brought up into the 500 A diameter class.

Association of xylem-limited bacteria with elm, sycamore, and oak leaf scorch

1980 ◽  
Vol 58 (18) ◽  
pp. 1986-1993 ◽  
Author(s):  
Suzanne S. Hearon ◽  
James L. Sherald ◽  
Stanley J. Kostka
Keyword(s):  
Fluorescent Antibody ◽  
Pierce's Disease ◽  
Tracheary Elements ◽  
Serological Relationship ◽  
Ulmus Americana ◽  
Leaf Scorch ◽  
Pierce’S Disease ◽  
Antibody Staining ◽  
Transmission Electron ◽  
Ultrathin Sections

Ultrathin sections of leaves from American elms (Ulmus americana L.), sycamores (Platanus spp.), and two red oak species (Quercus spp.) that exhibited leaf scorch were examined by transmission electron microscopy. Rod-shaped, ripple-walled bacteria resembling the Pierce's disease organism were found consistently in tracheary elements of the primary and secondary veins of diseased plants. Smaller, ripple-walled, densely stained, irregular-shaped bodies (SDB) were found also in a matrix that lined the inner walls or filled the lumina of the tracheary elements. In leaf scorch affected American elms the bacteria were 0.3–0.4 μrn × 0.9–2.4 μm, with rounded ends. Fimbriae-like structures radiated from a few organisms. Bacteria were frequently embedded in a matrix. Pit cavities and the ends of tracheary elements were often filled with the bacteria–matrix complex. Rod-shaped bacteria were not as numerous in the diseased sycamore and oak as in diseased elm; however, SDB's were more numerous. Bacteria in sycamore (1.0–1.8 μm) and oak (1.0–2.0 μm) were slightly shorter than those in elm, many had numerous fimbriae-like hairs, and some were tapered at one end. Indirect fluorescent antibody staining showed a serological relationship between bacteria extracted from elm and oak and the Pierce's disease bacterium.

Cytochemical localization of ATPase activity in the leaves of a seagrass and a submerged halophyte

1995 ◽  
Vol 53 ◽  
pp. 1058-1059
Author(s):  
A.D. Barnabas ◽  
M. Coopoosamy
Keyword(s):  
Atpase Activity ◽  
Epidermal Cells ◽  
Ruppia Maritima ◽  
Tolerance Mechanisms ◽  
Cytochemical Localization ◽  
X Ray ◽  
Halophila Ovalis ◽  
Transmission Electron ◽  
Ultrathin Sections ◽  
Edx Microanalysis

Flowering plants that grow submerged in seawater are known as seagrasses, while those that grow submerged in water of generally lower and varying salinity as in estuaries, are known as submerged halophytes. Knowledge of salt tolerance mechanisms in both groups of plants is important to our understanding of their biology. Recently, high ATPase activity was reported to be associated with the copiously-invaginated plasma membrane of leaf epidermal cells of a seagrass (Zostera marina), suggesting that ATPase played a role in salt regulation. In the present study, the seagrass Halophila ovalis and the submerged halophyte Ruppia maritima growing at salinities of 35°/00 and 25°/00 respectively, were used. The subcellular distribution of ATPase in leaf epidermal cells of both submerged aquatics was accomplished using a lead precipitation technique specific for ATPase activity. Lead deposits in the cells were an indication of sites of ATPase activity. Transmission electron microscopy (TEM) and energy dispersive X-ray (EDX) microanalysis using a Joel 6100 SEM with a Noran Voyager 2100 EDX microanalyser, were used to study lead distribution. Ultrathin sections for the TEM were examined unstained.

Platinum Compounds as Stains for Electron Microscopy

1974 ◽  
Vol 32 ◽  
pp. 230-231
Author(s):  
S. K. Aggarwal ◽  
P. McAllister ◽  
R. W. Wagner ◽  
B. Rosenberg
Keyword(s):  
Electron Microscopy ◽  
Hela Cells ◽  
Uranyl Acetate ◽  
Sarcoma 180 ◽  
Platinum Compounds ◽  
Kb Cells ◽  
En Bloc ◽  
Human Lymphoma ◽  
Ultrathin Sections ◽  
Blue Coloration

Uranyl acetate has been used as an electron stain for en bloc staining as well as for staining ultrathin sections in conjunction with various lead stains (Fig. 1). Present studies reveal that various platinum compounds also show promise as electron stains. Certain platinum compounds have been shown to be effective anti-tumor agents. Of particular interest are the compounds with either uracil or thymine as one of the ligands (cis-Pt(II)-uracil; cis-Pt(II)-thymine). These compounds are amorphous, highly soluble in water and often exhibit an intense blue coloration. These compounds show enough electron density to be used as stains for electron microscopy. Most of the studies are based on various cell lines (human AV, cells, human lymphoma cells, KB cells, Sarcoma-180 ascites cells, chick fibroblasts and HeLa cells) while studies on tissue blocks are in progress.

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.

Hemoglobin crystals of the red blood cells in the human renal cortex: electron microscopic observations of the renal lithiasis

1978 ◽  
Vol 36 (2) ◽  
pp. 480-481
Author(s):  
Kosuke Ueda ◽  
Hiroto Washida ◽  
Nakazo Watari
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Renal Cortex ◽  
Uranyl Acetate ◽  
Osmic Acid ◽  
Renal Lithiasis ◽  
Electron Microscopic ◽  
Hemoglobin C ◽  
First Case ◽  
Ultrathin Sections

IntroductionHemoglobin crystals in the red blood cells were electronmicroscopically reported by Fawcett in the cat myocardium. In the human, Lessin revealed crystal-containing cells in the periphral blood of hemoglobin C disease patients. We found the hemoglobin crystals and its agglutination in the erythrocytes in the renal cortex of the human renal lithiasis, and these patients had no hematological abnormalities or other diseases out of the renal lithiasis. Hemoglobin crystals in the human erythrocytes were confirmed to be the first case in the kidney.Material and MethodsTen cases of the human renal biopsies were performed on the operations of the seven pyelolithotomies and three ureterolithotomies. The each specimens were primarily fixed in cacodylate buffered 3. 0% glutaraldehyde and post fixed in osmic acid, dehydrated in graded concentrations of ethanol, and then embedded in Epon 812. Ultrathin sections, cut on LKB microtome, were doubly stained with uranyl acetate and lead citrate.

Albumins as Embedding Media for Electron Microscopy

1969 ◽  
Vol 27 ◽  
pp. 422-423 ◽  
Author(s):  
J. L. Farrant ◽  
J. D. McLean
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
High Temperature ◽  
Biological Material ◽  
Globular Proteins ◽  
The Past ◽  
Antibody Staining ◽  
Thin Sectioning ◽  
Embedding Medium

For electron microscope techniques such as ferritin-labeled antibody staining it would be advantageous to have available a simple means of thin sectioning biological material without subjecting it to lipid solvents, impregnation with plastic monomers and their subsequent polymerization. With this aim in view we have re-examined the use of protein as an embedding medium. Gelatin which has been used in the past is not very satisfactory both because of its fibrous nature and the high temperature necessary to keep its solutions fluid. We have found that globular proteins such as the serum and egg albumins can be cross-linked so as to yield blocks which are suitable for ultrathin sectioning.

Effects of Vibration on the Preparation of Ultrathin Sections

1973 ◽  
Vol 31 ◽  
pp. 358-359
Author(s):  
Joseph M. Blum ◽  
Edward P. Gargiulo ◽  
J. R. Sawers
Keyword(s):  
Cutting Speed ◽  
Ground Level ◽  
Environmental Vibration ◽  
Block Face ◽  
Ultrathin Sections ◽  
Embedding Medium ◽  
Thickness Variations ◽  
Building Walls ◽  
Cutting Direction ◽  
Diamond Knife

It is now well-known that chatter (Figure 1) is caused by vibration between the microtome arm and the diamond knife. It is usually observed as a cyclical variation in “optical” density of an electron micrograph due to sample thickness variations perpendicular to the cutting direction. This vibration might be induced by using too large a block face, too large a clearance angle, excessive cutting speed, non-uniform embedding medium or microtome vibration. Another prominent cause is environmental vibration caused by inadequate building construction. Microtomes should be installed on firm, solid floors. The best floors are thick, ground-level concrete pads poured over a sand bed and isolated from the building walls. Even when these precautions are followed, we recommend an additional isolation pad placed on the top of a sturdy table.

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