Catalytic Effect of Polyoxyethylene t -octylphenol (Triton X-100) Surfactant on the Oxidation of EDTA by Chromic Acid

The response of an uncatalyzed bromate oscillator with phenol as substrate to the increasing concentrations of cationic (CTAN), anionic (SDS) and nonionic surfactants (Brij-30 and Triton X-100) was monitored at (25±0.1) °C under stirred batch conditions. Addition of the surfactants influenced the oscillatory parameters: a slight increase of the induction period of the first series of oscillations, a significant increase of the induction period of the second series of oscillations and a gradual decrease of the oscillation numbers of both series until complete disappearance at a certain surfactant concentration. The changes in the oscillatory parameters have been ascribed to solubilization of phenol and of bromination products in the micelles, to inhibition of bromination of the aromatic substrate due to bromine solubilization, and to the catalytic effect of the charged micelle surface.

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Comparison of Acrylamide and Agar Gels by Freeze-Etching

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100080456 ◽

1977 ◽

Vol 35 ◽

pp. 606-607

Author(s):

Russell L. Steere ◽

Eric F. Erbe

Keyword(s):

Electron Microscope ◽

Sodium Hydroxide ◽

Sodium Hypochlorite ◽

Chromic Acid ◽

Distilled Water ◽

Thin Sheets ◽

Acid Cleaning ◽

Agar Gels ◽

Freeze Etching ◽

Water Cut

Thin sheets of acrylamide and agar gels of different concentrations were prepared and washed in distilled water, cut into pieces of appropriate size to fit into complementary freeze-etch specimen holders (1) and rapidly frozen. Freeze-etching was accomplished in a modified Denton DFE-2 freeze-etch unit on a DV-503 vacuum evaporator.* All samples were etched for 10 min. at -98°C then re-cooled to -150°C for deposition of Pt-C shadow- and C replica-films. Acrylamide gels were dissolved in Chlorox (5.251 sodium hypochlorite) containing 101 sodium hydroxide, whereas agar gels dissolved rapidly in the commonly used chromic acid cleaning solutions. Replicas were picked up on grids with thin Foimvar support films and stereo electron micrographs were obtained with a JEM-100 B electron microscope equipped with a 60° goniometer stage.Characteristic differences between gels of different concentrations (Figs. 1 and 2) were sufficiently pronounced to convince us that the structures observed are real and not the result of freezing artifacts.

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Chromic Acid Etching of Polyethylene

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100070801 ◽

1973 ◽

Vol 31 ◽

pp. 72-73

Author(s):

J. D. Muzzy ◽

R. D. Hester ◽

J. L. Hubbard

Keyword(s):

Chromic Acid ◽

Low Cost ◽

Acid Etching ◽

Crystalline Morphology ◽

Bulk Specimen ◽

Morphological Studies ◽

Perfect Form ◽

Molecular Length ◽

Lamellar Texture ◽

Cost Studies

Polyethylene is one of the most important plastics produced today because of its good physical properties, ease of fabrication and low cost. Studies to improve the properties of polyethylene are leading to an understanding of its crystalline morphology. Polyethylene crystallized by evaporation from dilute solutions consists of thin crystals called lamellae. The polyethylene molecules are parallel to the thickness of the lamellae and are folded since the thickness of the lamellae is much less than the molecular length. This lamellar texture persists in less perfect form in polyethylene crystallized from the melt.Morphological studies of melt crystallized polyethylene have been limited due to the difficulty of isolating the microstructure from the bulk specimen without destroying or deforming it.

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Freeze-etch studies of epiphyseal growth plate cartilage reveal matrix vesicles associated with calcification

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100084223 ◽

1978 ◽

Vol 36 (2) ◽

pp. 670-671

Author(s):

Russell N. A. Cecil ◽

H. Clarke Anderson

Keyword(s):

Chromic Acid ◽

Light And Electron Microscopy ◽

Matrix Vesicles ◽

Matrix Vesicle ◽

Glycerol Solution ◽

Sprague Dawley ◽

Epiphyseal Growth Plate ◽

Growth Plates ◽

Sprague Dawley Rats ◽

Tibial Epiphyseal

Unfixed proximal tibial epiphyseal growth plates were studied by freeze-etch to confirm the presence of extracellular calcifying matrix vesicles and to determine the substructure of matrix vesicle membranes as compared to plasma and other membranes of intact chondrocytes. Growth plates from 6-10 week old Sprague-Dawley rats were cut into 1x3 mm blocks whose long dimension was oriented either perpendicular or parallel to the long axis of the tibia. Some blocks were fixed at pH 7. 0 in 0. 2M cacodylate - buffered 2. 5% glutaraldehyde for 1 hour at 4ÅC. The blocks were immersed in 30% glycerol solution at 4ÅC for 1 hour, frozen in liquid nitrogen, and then fractured, etched for 2 minutes, and coated with platinum, carbon and 0. 2% Formvar solution. The replicas were cleaned with chromic acid, floated onto Formvar coated grids, and examined with a Phillips EM 300 electron microscope.Fixed and unfixed specimens appeared similar in ultrastructure. Chondrocytes, matrix, and matrix vesicles were identified. In specimens fractured parallel to the long axis of the tibia, the reserve, proliferative, hypertrophic, and calcifying zones could be discerned as described by light and electron microscopy.

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Demonstration of Retinal Glycogen with Silver Methenamine

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100066875 ◽

1971 ◽

Vol 29 ◽

pp. 564-565

Author(s):

A. W. Sedar ◽

G. H. Bresnick

Keyword(s):

Lactic Acid ◽

Chromic Acid ◽

Periodic Acid ◽

Müller Cell ◽

External Limiting Membrane ◽

Electron Microscope Level ◽

Reactive Process ◽

Inner Limiting Membrane ◽

Muller Cell ◽

And Migration

After experimetnal damage to the retina with a variety of procedures Müller cell hypertrophy and migration occurs. According to Kuwabara and others the reactive process in these injuries is evidenced by a marked increase in amount of glycogen in the Müller cells. These cells were considered originally supporting elements with fiber processes extending throughout the retina from inner limiting membrane to external limiting membrane, but are known now to have high lactic acid dehydrogenase activity and the ability to synthesize glycogen. Since the periodic acid-chromic acid-silver methenamine technique was shown to demonstrate glycogen at the electron microscope level, it was selected to react with glycogen in the fine processes of the Müller cell that ramify among the neural elements in various layers of the retina and demarcate these cells cytologically. The Rhesus monkey was chosen as an example of a well vascularized retina and the rabbit as an example of a avascular retina to explore the possibilities of the technique.

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High Resolution Stereography of Complementary Freeze-Fracture Replicas Reveals the Presence of Depressions Opposite Membrane Associated Particles of Split Membranes

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100066267 ◽

1971 ◽

Vol 29 ◽

pp. 442-443

Author(s):

Russell L. Steere

Keyword(s):

High Resolution ◽

Cardiac Muscle ◽

Electron Micrographs ◽

Chromic Acid ◽

Freeze Fracture ◽

Distilled Water ◽

Fine Scale ◽

Acid Cleaning ◽

Cleaning Solution

Complementary replicas have revealed the fact that the two common faces observed in electron micrographs of freeze-fracture and freeze-etch specimens are complementary to each other and are thus the new faces of a split membrane rather than the original inner and outer surfaces (1, 2 and personal observations). The big question raised by published electron micrographs is why do we not see depressions in the complementary face opposite membrane-associated particles? Reports have appeared indicating that some depressions do appear but complementarity on such a fine scale has yet to be shown.Dog cardiac muscle was perfused with glutaraldehyde, washed in distilled water, then transferred to 30% glycerol (material furnished by Dr. Joaquim Sommer, Duke Univ., and VA Hospital, Durham, N.C.). Small strips were freeze-fractured in a Denton Vacuum DFE-2 Freeze-Etch Unit with complementary replica tooling. Replicas were cleaned in chromic acid cleaning solution, then washed in 4 changes of distilled water and mounted on opposite sides of the center wire of a Formvar-coated grid.

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SEM of non-coated hepatocellular cytoskeleton of perfused livers

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100111641 ◽

1984 ◽

Vol 42 ◽

pp. 306-307

Author(s):

S.W. French ◽

N.C. Benson ◽

C. Davis-Scibienski

Keyword(s):

Ambient Temperature ◽

Critical Point ◽

Protease Inhibitors ◽

Metal Deposition ◽

Heat Damage ◽

Detergent Extraction ◽

Cytoskeletal Elements ◽

Triton X ◽

Excised Tissue

Previous SEM studies of liver cytoskeletal elements have encountered technical difficulties such as variable metal coating and heat damage which occurs during metal deposition. The majority of studies involving evaluation of the cell cytoskeleton have been limited to cells which could be isolated, maintained in culture as a monolayer and thus easily extracted. Detergent extraction of excised tissue by immersion has often been unsatisfactory beyond the depth of several cells. These disadvantages have been avoided in the present study. Whole C3H mouse livers were perfused in situ with 0.5% Triton X-100 in a modified Jahn's buffer including protease inhibitors. Perfusion was continued for 1 to 2 hours at ambient temperature. The liver was then perfused with a 2% buffered gluteraldehyde solution. Liver samples including spontaneous tumors were then maintained in buffered gluteraldehyde for 2 hours. Samples were processed for SEM and TEM using the modified thicarbohydrazide procedure of Malich and Wilson, cryofractured, and critical point dried (CPD). Some samples were mechanically fractured after CPD.

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Immunoelectrophoretic Analysis of Membrane Glycoproteins in Cultured Human Endothelial Cells

Thrombosis and Haemostasis ◽

10.1055/s-0038-1645214 ◽

1990 ◽

Vol 63 (02) ◽

pp. 303-311

Author(s):

Tone Børsum

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

Rabbit Antiserum ◽

Platelet Glycoprotein ◽

Membrane Glycoproteins ◽

Human Endothelial Cells ◽

Immunoelectrophoretic Analysis ◽

Glycoprotein Complex ◽

Crossed Immunoelectrophoresis ◽

Triton X

SummaryHuman endothelial cells isolated from umbilical cordswere solubilized in Triton X-100 and examined by crossedimmunoelec-trophoresis using rabbit antiserum against endothelial cells. Endogenous labelling of the endothelialcell proteins with 14Cmannose followed by crossed immunoelectrophoresis and autoradiography revealed about 10 immunoprecipitates. Four of these endothelial cell glycoproteins were labelled by lactoperoxidase catalyzed iodination and thus were surface located. Three of the surface located glycoproteins showed reduced electrophoretic mobility after incubation of the endothelial cells with neuraminidase and were therefore sialoglycoproteins. Amphiphilicity of endothelial cell glycoproteins was studied by crossed hydrophobic interaction immunoelectrophoresis with phenyl-Sepharose in the intermediate gel. Amphiphilic proteins also show increasing electrophoretic migration velocity with decreasing concentration of Triton X-100 in the first dimension gels. Five of the endothelial cell glycoproteins were shown to be amphiphilic using these two techniques.Two monoclonal antibodies against the platelet glycoprotein complex Ilb-IIIa and glycoprotein IlIa, respectively, reacted with the same precipitate of endothelial cells. When a polyclonal antibody against the platelet glycoprotein complex Ilb-IIIa was incorporated into the intermediate gel the position of two endothelial cell precipitates were lowered. One of these was a sialoglycoprotein.

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The Uptake of Adenine by Isolated Human Platelet Membranes

Thrombosis and Haemostasis ◽

10.1055/s-0038-1647713 ◽

1974 ◽

Vol 32 (02/03) ◽

pp. 457-464

Author(s):

Paul C. French ◽

Jan J. Sixma ◽

Holm Holmsen

Keyword(s):

Human Platelet ◽

Facilitated Diffusion ◽

Adenine Phosphoribosyltransferase ◽

Ph Optimum ◽

Intact Cells ◽

Platelet Membranes ◽

Group Translocation ◽

Triton X

SummaryAdenine uptake into isolated platelet membranes had about the same Km (151 ± 21 • 9 nM) as uptake into intact cells (159 ± 21 nM) and was also competitively inhibited by papaverine and hypoxanthine. No uptake occurred at 0° and accumulated adenine was converted to AMP. AMP was not firmly bound to protein as judged by chromatography of triton X-100 solubilized membranes on Sephadex G25. The pH optimum for adenine uptake was at pH 5-5. Exogenous 5-phosphoribosyl-l-pyrophos- phate strongly stimulated uptake. These data may be explained by uptake of adenine by facilitated diffusion followed by conversion to AMP by adenine phosphoribosyltransferase but group translocation cannot be entirely excluded.

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