Showing papers on "Glutaraldehyde published in 1983"
TL;DR: Using these methods, specific immunological staining is achieved throughout the full thickness of retinal slices, up to 500 microns across, while preserving good ultrastructure and should prove useful in the immunocytochemical localization of many different antigens in a variety of tissues.
Abstract: Electron microscopic immunohistochemistry, although generally providing good localization, has often failed to produce satisfying ultrastructural preservation. Techniques that result in well-preserved tissue ultrastructure often hinder penetration of immunological reagents or render antigens non-immunoreactive. These are particularly serious limitations in studies of central nervous system and retina. We have evaluated several fixatives, including picric acid, high pH paraformaldehyde, and glutaraldehyde with subsequent sodium borohydride treatment, and penetration enhancement techniques, including buffered-ethanolic treatment and freeze--thaw, for their applicability in the retina. Our best fixation was achieved with 1 hr in 4% paraformaldehyde and 0.2% glutaraldehyde (pH 7.4) followed by overnight fixation in 4% paraformaldehyde (pH 10.4). Treatment with sodium borohydride after glutaraldehyde fixation restores much of the immunoreactivity that would otherwise be undetectable. The penetration of immunol...
353 citations
TL;DR: A chemically modified enzyme membrane electrode for glucose was constructed by cross-linking glucose oxidase with bovine serum albumin using glutaraldehyde onto a platinum electrode silanized with 3-aminopropyltriethoxysilane.
137 citations
TL;DR: The use and properties of an alternative fixation procedure for ultrastructural cytochemistry is described that utilized primary fixation of cells with glutaraldehyde with considerable latitude in concentration and time of fixation, and treatment with sodium borohydride increases the accessibility of some cytoplasmic compartments to antibodies.
Abstract: The use and properties of an alternative fixation procedure for ultrastructural cytochemistry is described that utilized primary fixation of cells with glutaraldehyde with considerable latitude in concentration and time of fixation. This was followed by treatment with sodium borohydride (as described by Weber, Rathke and Osborn (Proc Natl Acad Sci USA 75:1820, 1978)), which increases the accessibility of some cytoplasmic compartments to antibodies, apparently through the reduction of the Schiff bases induced by glutaraldehyde. Through the use of saponin membrane permeabilization, this primary fixation method allows good ultrastructural preservation with accessibility of most, but not all, cytoplasmic structures. Localization of tubulin, clathrin, alpha2-macroglobulin present in lysosomes, vesicular stomatitis virus (VSV) G protein, and myosin are demonstrated. This technique failed to expose the antigenic structure of actin in microfilament bundles or intranuclear antigens. This method should be useful fo...
127 citations
Patent•
04 Jul 1983TL;DR: In this paper, a solution of a water soluble phosphate ester such as sodium dodecyl hydrogen phosphate was used to inhibit mineralization, particularly calcification, of the tissue after implantation.
Abstract: Natural tissues fixed with a tanning solution such as glutaraldehyde and intended for implantation in humans, e.g., porcine heart valve prosthetic devices, are treated with a solution of a water soluble phosphate ester such as sodium dodecyl hydrogen phosphate to inhibit mineralization, particularly calcification, of the tissue after implantation.
121 citations
TL;DR: The aim of the present investigation was to study the extent to which lipids are extracted from biological membranes during dehydration and embedding procedures carried out at high or low temperatures.
Abstract: SUMMARY
The aim of the present investigation was to study the extent to which lipids are extracted from biological membranes during dehydration and embedding procedures carried out at high or low temperatures. Cells of Acholeplasma laidlawii were used as experimental material, since the lipids of this bacterium easily can be radioactively labelled without labelling the rest of the cell, and the lipids are almost entirely located in the cytoplasmic membrane. The cells were fixed at 277 K with glutaraldehyde, sequentially with this reagent and osmium tetroxide, or with glutaraldehyde, osmium tetroxide and uranyl acetate in that order. Loss of lipid during these procedures was negligible.
When cells fixed with glutaraldehyde and osmium tetroxide were dehydrated with ethanol at room temperature and embedded in Epon at 333 K, i.e. subjected to a conventional treatment, about 90% of the lipid content of the cells was extracted. The loss was reduced to c. 20% when treatment with uranyl acetate was included in the procedure and the non-polar methacrylate resin Lowicryl HM20 was substituted for Epon. When cells fixed with glutaraldehyde and osmium tetroxide were dehydrated with ethanol at 238 K and embedded in Lowicryl HM20 at room temperature, practically no lipid was extracted. Substitution of the polar methacrylate-acrylate resin Lowicryl K4M for Lowicryl HM20 resulted in the loss of about half of the lipid content of the cells. The use of ethanediol as dehydrating agent instead of ethanol did not diminish the extraction. Cells fixed solely with glutaraldehyde lost about half of their lipid content, even when both dehydration and embedding was performed at 238 K.
The lipid material extracted from glutaraldehyde-fixed cells contained slightly more saturated fatty acids than that remaining in the cells. The reverse was true for osmium tetroxide-fixed cells. With respect to lipid species, the extractions were generally rather unspecific.
90 citations
TL;DR: It is concluded that with proper fixation immune cell surface markers can be identified in paraffin-embedded mouse lymphoid tissue.
90 citations
TL;DR: The quaternARY ammonium compound as well as the quaternary ammonium-glutaraldehyde complex were more readily neutralized whereas anionic acid, iodophor and sodium hypochlorite did not tolerate the presence of organic matter.
Abstract: The effect of organic matter on the activity of eight disinfectants was evaluated. Three types of interfering substrates (whole milk powder, dried beef blood and fish meal) were tested according to the method of Whitmore and Miner adapted to the AOAC use-dilution method. Glutaraldehyde and to a certain extent, chlorhexidine acetate and the amphoteric surfactant kept their disinfecting activity after contact with high concentrations of organic matter. The quaternary ammonium compound as well as the quaternary ammonium-glutaraldehyde complex were more readily neutralized whereas anionic acid, iodophor and sodium hypochlorite did not tolerate the presence of organic matter. The neutralizing activity of powders was correlated to their solubility and composition.
89 citations
TL;DR: It is indicated that glutaraldehyde enhancement of protein staining with this silver reagent was probably due to oxidation of the aldehyde groups by silver ions, resulting in metallic silver depositions within the gel which act as nucleation sites for additional silver localization in the protein bands upon the addition of formaldehyde developer.
75 citations
TL;DR: Lipid extraction prior to lectin incubation resulted in complete elimination of detectable binding to epithelium suggesting that lectin-binding sites in the cell surface are associated with glycolipid or lipid.
Abstract: The effects of fixation and wax processing on lectin binding to C3H mouse palate and tail skin were evaluated using eight FITC-conjugated lectins. Sections and blocks of tissue were fixed in acetone, ethanol, methanol, formalin, glutaraldehyde or Bouin's picric-acetic-formalin fixative. Tissue blocks were then processed to paraffin wax.
63 citations
TL;DR: The effect of treating the soil with sporicidal chemicals, namely, potassium permanganate, formaldehyde, glutaraldehyde, Cidex, Surgikos, dodecylamine and peracetic acid, is investigated.
Abstract: In experiments on Gruinard Island 40 years ago small bombs containing spores of Bacillus anthracis were suspended from a gantry and detonated, producing widespread contamination of the island's surface. Recently, analysis of soil samples has shown that the area where the spores can now be detected is small enough to be considered for decontamination1. We investigated the effect of treating the soil with sporicidal chemicals, namely, potassium permanganate, formaldehyde, glutaraldehyde, Cidex (‘activated’ glutaraldehyde, Surgikos), dodecylamine and peracetic acid.
56 citations
TL;DR: Anion exchange membranes containing amino groups, insoluble in acidic and alkaline aqueous solutions, were prepared from chitosan, poly(vinyl alcohol), and glutaraldehyde to transport actively halogen ions through the membrane.
Abstract: Anion exchange membranes containing amino groups, insoluble in acidic and alkaline aqueous solutions, were prepared from chitosan, poly(vinyl alcohol), and glutaraldehyde. Using the membrane in a diaphragm cell, one side being adjusted to be acidic and the other alkaline, it was possible to transport actively halogen ions through the membrane from the acidic side to the alkaline side against the concentration gradient between both sides of the membrane. The active transport of halogen ions through the membrane was significantly influenced by the pH difference and electric potential difference between both sides of the membrane.
Journal Article•
TL;DR: The cytotoxicity of aldehyde-treated collagen was assayed by measuring 3H-thymidine incorporation in adult human skin fibroblasts grown in tissue culture for 1 or 3 days in the presence of pig dermal collagen cross-linked with formaldehyde or glutaraldehyde.
Abstract: The cytotoxicity of aldehyde-treated collagen was assayed by measuring 3H-thymidine incorporation in adult human skin fibroblasts grown in tissue culture for 1 or 3 days in the presence of pig dermal collagen cross-linked with formaldehyde or glutaraldehyde. A comparison was also made with collagen preparations washed for 2 weeks either at 15 degrees throughout or partly at 15 degrees and partly at 37 degrees. Collagen treated with both formaldehyde and glutaraldehyde proved increasingly toxic with increase in the concentrations of aldehyde used. While the maximum toxic effect was observed after 1 day culture in formaldehyde-treated collagen, with thymidine uptake ranging from 4-48% of control values with 5-0.1% formaldehyde and a 15 degrees wash, the toxic effect of glutaraldehyde treatment increased with longer exposure and at 3 days thymidine uptake ranged from 3-40% of control values with 0.05-0.001% glutaraldehyde and washing at 15 degrees. Washing partly at 37 degrees significantly reduced toxicity, the differences in thymidine uptake as compared with washing at 15 degrees alone ranging from 34-50% with 1 and 0.3% formaldehyde respectively in 1 day cultures and from 14-37% with 0.02 and 0.005% glutaraldehyde in 3 day cultures. While fibroblasts actively grew and migrated when seeded on non-cross-linked collagen, only limited cell survival occurred on aldehyde-treated collagen.
TL;DR: Fresh and stabilised reagent-linked cells were shown to compare favourably in reverse passive haemagglutination for the measurement of human immunoglobulin isotypes, G, A and M and for the detection of respiratory syncytial and herpes simplex viruses.
TL;DR: Within a range of biologically active concentrations, glutaraldehyde did not produce significant genotoxic effects with the assays and conditions used for these studies.
TL;DR: It is proposed that glutaraldehyde stabilizes the ADH-induced channels by cross-linkage of amino groups and other reactive sites at the cytoplasmic surface of the apical membrane and/or by inactivating the intracellular machinery responsible for the dispersal or removal of water channels in the hormone target cell.
Abstract: The present study investigates the time-, dose-, and temperature-dependence of glutaraldehyde action on the permeability to water of the toad bladder. Bladders preincubated with increasing concentrations of glutaraldehyde become progressively desensitized to the hydrosmotic action of vasopressin (ADH), theophylline, and dibutyryl adenosine 3',5'-cyclic monophosphate (dibutyryl cAMP). The ADH response was reduced by 50% with 0.03% glutaraldehyde applied to the serosal side for 10 min at 4 degrees C. Sixfold higher doses of glutaraldehyde were required with mucosal application. Bladders partially fixed with low-dose glutaraldehyde exhibit a markedly prolonged duration of action of ADH. Bladders fixed with higher doses of glutaraldehyde in the presence of ADH retain a high permeability to water for prolonged periods even in the absence of ADH. This action of glutaraldehyde to stabilize the hormone-induced water channels is also considerably more effective with serosal than with mucosal application. As the ra...
TL;DR: Two liquid glutaraldehyde preparations and a 5.25% aqueous sodium hypochlorite solution were compared for effectiveness in sterilizing gutta-percha cones artificially contaminated with Bacillus subtilis spores.
Patent•
21 Nov 1983
TL;DR: In this paper, a polyvinyl alcohol-aldehyde reaction product is produced continuously in a rotating contactor by contacting solid particles of the alcohol with the aldehyde in an acidic aqueous salt solution.
Abstract: A polyvinyl alcohol-aldehyde reaction product is produced continuously in a rotating contactor by contacting solid particles of the polyvinyl alcohol with the aldehyde in an acidic aqueous salt solution. The product can be dried and then sieved to remove oversize particles. A preferred aldehyde is either formaldehyde or glutaraldehyde which reacts with the polyvinyl alcohol to produce a reaction product that can be used as a fluid loss control agent in oil fields.
TL;DR: Treatment of the conjugate with urea resulted in a 20--30-fold increase in specific activity, suggesting that a number of chelon groups are unavailable for Ga binding due to non-covalent intramolecular cross-linking.
01 Jul 1983
TL;DR: The 77 K emission spectra was however found to be the same in free and in algae immobilized in the serum albumin glutaraldehyde matrix, suggesting that the electron transfer from Q to plastoquinone is altered in this case.
Abstract: Cells of the green microalgae Scenedesmus obliquus have been immobilized in alginate or serum albumin glutaraldehyde matrices. The alginate matrix conserved the properties of the algae (photosynthetic activity and fluorescence characteristics). On the contrary, the serum albumin glutaral dehyde matrix inhibited the photosynthetic activity, and modified the kinetic of the room temperature 690 nm fluorescence. This suggests that the electron transfer from Q to plastoquinone is altered in this case. The 77 K emission spectra was however found to be the same in free and in algae immobilized in the serum albumin glutaraldehyde matrix.
TL;DR: Glutaraldehyde is proposed as an alternative to the more noxious hypochlorite and formaldehyde solutions for disinfection of HBV-contaminated articles.
Abstract: The potential of alkaline 2% glutaraldehyde solutions, with and without surface active agents, to alter the antigenicity of hepatitis B virus (HBV) was analyzed and compared to the antigenic alternation capacities of 0.525% sodium hypochlorite and 2.02% formaldehyde solutions. After treatment of a hepatitis B surface antigen-positive plasma at room temperature for 10 min, there was a 51-67% reduction in surface antigen level and a 90-94% decrease in hepatitis B core antigenicity. Glutaraldehyde is proposed as an alternative to the more noxious hypochlorite and formaldehyde solutions for disinfection of HBV-contaminated articles.
TL;DR: Two bifunctional reagents, glutaraldehyde and dimethylsuberimidate, were compared to formaldehyde with respect to fixation and modification of protein and an immunoprecipitation assay suggests that the reaction products of glutarhyde may have altered antigenicity.
Abstract: Two bifunctional reagents, glutaraldehyde and dimethylsuberimidate, were compared to formaldehyde with respect to fixation and modification of protein. Glutaraldehyde proved the superior fixative based on cross-linking and enzyme degradation assays. An immunoprecipitation assay suggests that the reaction products of glutaraldehyde may have altered antigenicity.
TL;DR: Crosslinkage of anti-human albumin with varying concentrations of glutaraldehyde to Staphylococcus aureus Cowan 1 (SpA-Staph) and to stAPHylococcal protein A-Sepharose ( SpA-SepHarose) was tested and antibody activity was more than 60 and 90% respectively compared with the corresponding noncrosslinked immunosorbents.
Journal Article•
TL;DR: Polyheads of bacteriophage T4 are in dynamic equilibrium with their subunits and fit very well as a probe to measure the efficiency of crosslinking by the arrest of dissociation upon dilution and upon treatment with hot SDS.
TL;DR: In this article, an endopolygalacturonase was immobilized by coupling on to porous poly(2,6-dimethyl-p-phenyleneoxide) activated by adsorbed glutaraldehyde.
Abstract: Endopolygalacturonase was immobilized by coupling on to porous poly(2,6-dimethyl-p-phenyleneoxide) activated by adsorbed glutaraldehyde. Catalytic properties, stability and action pattern of the immobilized enzyme are described.
TL;DR: Vascular reactions to fixation with a 2·5% w/v glutaraldehyde fixative were studied in a constant flow perfusion system on isolated rabbit intestines and hind limb preparations to assess changes in vascular resistance and macro‐molecular permeability.
Abstract: Vascular reactions to fixation with a 2 x 5% w/v glutaraldehyde fixative were studied in a constant flow perfusion system on isolated rabbit intestines and hind limb preparations. Reactions induced by the fixative were compared to reactions induced by noradrenaline in each single preparation. Changes in vascular resistance were assessed from continuous recordings of arterial and venous pressures and perfusion flow. Changes in vascular filtration and macromolecular permeability during fixation were approximated from recordings of flow and measurements of concentrations of dextran in the perfusate at the arterial inlet and at the venous outlet. Our results indicate that the vascular bed of the preparations studied can be reliably fixed in a high resistance state of smooth muscle activity induced by noradrenaline as well as in a low resistance state in the absence of noradrenaline, provided that hydrodynamic parameters are strictly controlled. The vascular filtration decreases during fixation. In optimally perfused preparations 5-10 min are required to fully saturate the binding capacity of the tissue for glutaraldehyde. Data on viscosity and colloid osmotic pressure of Tyrode solution containing dextran T-70 in various concentrations, and of various aldehyde-containing fixatives in 0 . 1 M sodium phosphate buffer with and without dextran added, are given.
TL;DR: A triple-fixation method with a sequential application of 5% glutaraldehyde, 1% osmium tetroxide, and 2% potassium permanganate gave superior preservation of the ultrastructure of Bacillus subtilis dormant spores with a thick spore coat.
Abstract: A triple-fixation method with a sequential application of 5% glutaraldehyde, 1% osmium tetroxide, and 2% potassium permanganate gave superior preservation of the ultrastructure of Bacillus subtilis dormant spores with a thick spore coat.
TL;DR: The possible role of exocytosis as a mode of secretion in the glomus cells and the characteristics of the new high K-glutaraldehyde fixative are discussed.
Abstract: Extensive secretion by exocytosis was demonstrated in the glomus (type I) cells of the adult rat after perfusion of carotid bodies with a potassium-rich (high K) glutaraldehyde fixative. Similar secretory profiles were very rare with a glutaraldehyde fixative containing a low concentration of potassium (low K). The increase in the incidence of exocytotic profiles in glomus cells with the high K fixative was highly significant, whereas no statistical difference could be observed in the incidence of coated pits with the different fixatives. Exocytotic profiles were characterized by the following features: (1) they predominated in non-synaptic regions, but were occasionally observed near synapses between two glomus cells; they were not observed near synapses between glomus cells and nerve terminals; (2) extruded electron-dense material associated with coating of the cell membrane was frequent; (3) different stages of dissolution of the extruded granule material was evident. The possible role of exocytosis as a mode of secretion in the glomus cells and the characteristics of the new high K-glutaraldehyde fixative are discussed.
TL;DR: The assay was sensitive and reproducible, and lent itself to the simultaneous evaluation of many individual antibody samples in a short period of time, and was particularly valuable for rapid screening of hybridoma supernatants for antibodies to antigens derived from melanoma cells and from a panel of other tumor and normal cells.
Abstract: A solid-phase radioimmunoassay to detect antibodies that react with antigens derived from human melanoma cells is described. A soluble preparation derived from Nonidet P-40 lysates of tissue-cultured melanoma cells was dried on the surfaces of wells of polyvinyl chloride microtiter plates and fixed with 0.02% glutaraldehyde. Antibody preparations were added and incubated for 18 h at 4° C. The wells were washed and bound antibodies were detected using radioactive Staphyloccoccal protein A (125I-SpA). Optimal conditions are described for all the steps employed. Concentrations of antigen selected, the amount of 125I-SpA employed and the duration of incubation of antibodies with antigen were found to be critical.