Showing papers on "Glutaraldehyde published in 1968"

Ultrastructure of human leukocytes after simultaneous fixation with glutaraldehyde and osmium tetroxide and "postfixation" in uranyl acetate.

Abstract: Human leukocytes in suspension or in monolayer cultures have been processed for electron microscopy by fixation in a freshly made cold mixture of glutaraldehyde and osmium tetroxide and by "postfixation" in uranyl acetate. Simultaneous exposure to glutaraldehyde and osmium tetroxide eliminates many of the shortcomings seen when either of these agents is used alone as the initial fixative. Specimens are processed to the stage of dehydration as single cell suspensions or as very small clumps to assure rapid penetration of fixatives and efficient washing. The technique is rapid and reproducible. Electron micrographs presented in this report illustrate the ultrastructural features of human white cells prepared by this method.

Alteration of the conformation of proteins in red blood cell membranes and in solution by fixatives used in electron microscopy.

Abstract: The effects of several commonly employed fixatives on the three-dimensional conformations of two soluble proteins and the protein of intact red blood cell membranes have been studied by means of circular dichroism measurements in the spectral region of the peptide absorption bands. The fixatives used produced significant and parallel conformational changes in all of the proteins, in the increasing order: glutaraldehyde; OsO4; glutaraldehyde followed by OsO4; and KMnO4. The last two treatments obliterated most of the helical character of the proteins. The significance of these observations to the preparation of specimens for electron microscopy is discussed.

Topical glutaraldehyde for plantar hyperhidrosis.

Abstract: Glutaraldehyde is a tanning agent with strong bactericidal and fungicidal properties. Topical application of a 10% solution was used with good effect against hyperhidrosis of the feet. No cross reaction was found in patients sensitized to formaldehyde.

Perfusion fixation with glutaraldehyde and post-fixation with osmium tetroxide for electron microscopy.

Abstract: The conductivity of cerebral cortex drops during perfusion with glutaraldehyde in 5 min to about 60% of the original value, to remain unchanged during the subsequent 10-15 min of perfusion. Circulatory arrest causes a similar drop in the tissue conductivity. Perfusion of asphyxiated tissue with glutaraldehyde does not produce additional major changes in the conductivity. Perfusion of the cortex with an osmium tetroxide solution causes an initial drop in conductivity. However, after about 3 min this trend is reversed and the conductivity increases again to close to the pre-perfusion value. Perfusion of asphyxiated cortex with OsO4 causes a marked increase of the conductivity. So does perfusion with an OsO4 solution of tissue previously treated with glutaraldehyde. One interpretation of these impedance changes is that glutaraldehyde perfusion causes, like asphyxiation, a transport of extracellular material into the intracellular compartment and that during OsO4 perfusion an extracellular space is again created. This possibility is supported by electron micrographs made of this material. Cerebral cortex perfused with glutaraldehyde and post-fixed with OsO4 shows electron-transparent dendritic elements and to a lesser extent pre-synaptic terminals, which seem to be swollen. When the cortex is flooded with a salt solution during glutaraldehyde perfusion the dendrites exhibit ballooning in the surface layer of the cortex, suggesting that the fluid on the cortex participates in the swelling. The tissue elements in the glutaraldehyde-perfused and OsO4 post-fixed cortex are separated by narrow extracellular spaces. The latter may have been produced by the OsO4 perfusion as is suggested by a comparison of micrographs prepared by freeze substitution (which tends to preserve the water distribution) of glutaraldehyde-perfused but not post-fixed cortex with micrographs of cortex treated with OsO4 after the glutaraldehyde perfusion. In accordance with the conductivity changes, the former micrographs showed very little extracellular space, and in many places tight junctions, whereas the latter showed clefts between the tissue elements.

Purification of glutaraldehyde its significance for preservation of acid phosphatase activity

Abstract: The inhibition of acid phosphatase activity in rat liver homogenates after fixation in different lots of commercial glutaraldehyde is determined and compared with the inhibition following fixation with a distilled product. It is shown that commercial glutaraldehydes inhibit more of the enzyme activity than the distilled product. The acidic products of oxidation of glutaraldehyde do not increase the Inhibition of the enzymatic activity. The presence of high concentration of inorganic phosphates in different lots of commercial glutaraldehyde, as presented here, suggests that probably such impurities may be responsible for Increased inhibition of phosphatase activity noted after fixation in commercial glutaraldehydes.

Electron microscope studies of glutamic oxalacetic transaminase in rat liver cell.

Abstract: Liver tissue of the rat, fixed in glutaraldehyde and formaldehyde, was incubated in a medium which consisted of 20 mM L-aspartic acid, 2 mM α-ketoglutaric acid, 50 mM imidazole and 6 mM lead nitrate at pH 7.2–7.4. The electron-opaque precipitates, due to glutamic oxalacetic transaminase activity in liver cells, were found to be localized to the cristae and surface membranes of the mitochondria, the limiting membrane of the microbodies, and the nuclear membrane. Sucrose storage and trauma resulted in altered morphology and diminished final product intensity in mitochondria, but the microbody enzyme activity disappeared completely under these conditions. These distinctive responses of enzymatic activity are considered to indicate a difference in either the enzyme protein or its membrane attachment to these two sites. The use of a buffered dehydrating ethanol series to prepare tissue blocks for electron microscopy appeared to result in more precise intracellular localization of enzymatic reaction product.

Feulgen Cytophotometry of Pine Nuclei; Effects of Fixation, Role of Formaline

Abstract: The effects of 5 fixatives: FAA, Carnoy's, Craf III, formalin and glutaraldehyde were analyzed for use in quantitative Feulgen cytophotometry of pine embryo tissues. Craf III and glutaraldehyde had serious deficiencies because they depressed the absorption peak, severely interfered with DNA extraction and in the case of glutaraldehyde there was considerable cytoplasmic dye-binding. Neutral 10% formalin gave good tissue fixation but did not permit the degree of enzymatic or acid extraction of DNA as did Carnoy's solution. Haupt's adhesive, with the usual 4% formalin as a hardener, at temperatures of 45-56 C completely prevented the enzymatic extraction of nuclear DNA by DNase and also greatly increased the resistance of the DNA to mineral acid hydrolysis. Denaturation of DNA by formalin appeared to be responsible for these results. Absorption was linearly related to both section thickness and DNA concentration per nucleus.

Aldehyde as fixative for histochemical study of glutamic oxaloacetic transaminase.

Abstract: Several compounds of the aldehyde group have been tested as fixatives for histochemical localization of GOT activity in the rat kidney. A brief fixation of cryostat tissue sections in 1% glutaraldehyde results in well localized enzymatic activity which is not greatly further affected by an additional treatment in 4% formaldehyde.

Chloride movements during perfusion fixation with glutaraldehyde.

Abstract: The cerebral and cerebellar cortices of mice were subjected to a histochemical method for the demonstration of chloride after perfusion fixation with glutaraldehyde. Unfixed, nonasphyxiated cortices subjected to the same chloride method were used as controls. Sections of control material showed a rather uniform chloride distribution in which only the pia and the blood vessels stood out as dark structures containing an appreciable amount of chloride. Sections of glutaraldehyde fixed cerebral cortex showed an accumulation of chloride in apical dendrites. In glutaraldehyde fixed cerebellar cortex the chloride accumulated in the fibers of Bergmann and sometimes in the large dendrites of Purkinje cells. The chloride movement during glutaraldehyde perfusion is in all respects comparable with the movement of chloride caused by asphyxiation of the cerebral and cerebellar cortex.

Syntheses of some branched-chain nitrocycloalkanols.

Abstract: Base-catalyzed cyclizations of glutaraldehyde with nitroethane, β-nitropropionic acid, β-nitro-α-methoxypropionic acid, and methyl vinyl ketone were investigated. The structures of these resulting nitrocycloalkanols were determined by NMR analysis and chemical reactions.

The organization of chloroplast membranes ofVicia faba andZea mays after processing to retain chlorophyll and hydrophobic lipids in the chloroplasts

Abstract: Chloroplasts, bothin situ and isolated, were dehydrated at low temperatures to avoid chlorophyll extraction. These chloroplasts were fixed with glutaraldehyde only or with glutaraldehyde followed by osmium. Chloroplasts dehydrated at low temperatures have narrower fret membranes, but the same dimensions of the partitions as chloroplasts dehydrated by standard techniques. However, the two membranes forming the partition are separated by an electron transparent region or A-component in low temperature dehydrated chloroplasts post-fixed with osmium. With glutaraldehyde only, the A-component appears as a thin discontinuous dark line. The results indicate that the A-component is a structural component of the partition and supports the hypothesis that chlorophyll and the hydrophibic lipids are localized there.

[Electron microscopical studies of differentiating processes in mosses : I. Problems of preserving the fine structure].

Abstract: For the demonstration of the ultrastructure of the cells from Riella and Funaria by electron microscopy, special conditions were found to be necessary for preservation of all known structural elements. The best results were obtained by fixation in a buffered mixture of formaldehyde (0.5 or 1.5%) and glutaraldehyde (0.5 or 1.5%) for 15-20 hours in the cold. The postfixation was performed in 1% OsO4 for 3 hours. Using these conditions an excellent preservation of all structural elements is demonstrated in young and old cells of the mosses. The conditions used in our experiments are compared with those from other experiments performed previously by other investigators who were not so successful in overcoming the special difficulties encountered in the study of mosses.

Moth Resistance of Glutaraldehyde-Stabilized Wool

Abstract: In any case, examination of the deviations observed (Fig. 3 in reference [1]) lead to maximum differences of the order of i0.07 in. (1.8 mm), which seems to be excessive from the point of view of industrial practice, which confirms the conclusions from a study of the present authors [3] where a warning is expressed on the dangers of estimating the parameters of the array from data of the Digital Fibrograph. It is evident that the technique proposed by Louis and Fiori is interesting, but its practical application seems to be subject to certain limitations; above’ all if there

Dialdehyde tanned edible collagen casing

Abstract: 1,122,505. Tubular collagen films. TEEPAK Inc. 18 April, 1966 [28 June, 1965], No. 16876/66. Heading B5B. A process of tanning an extruded edible casing comprising collagen fibrils comprises contacting said casing with a solution of an edible non-toxic dialdehyde at a pH of 4.0 to 7.0 and washing and drying the casing. The dialdehyde solution may contain a buffer salt. Preferably, the casing is contacted with a solution of glutaraldehyde or condensed smoke containing non-toxic dialdehydes in solution, e.g. the commercially available Griffith SF 12 " liquid smoke " (actually a condensed smoke) adjusted to a suitable pH, or alternatively, may be a solution of so-called " liquid smoke" made by passing smoke from sawdust or wood shavings burnt under controlled combustion conditions and passed through water, having a suitable pH. Suitably, the solution contains glutaraldehyde at a pH of 4 to 6, the solution desirably having a glutaraldehyde concentration of 1 to 50% by weight. In a preferred method an edible tubular collagen casing is made by defleshing an animal hide and removing the epidermal layer and hair therefrom, cutting the hide into pieces, grinding the hide pieces at a temperature less than 20‹ C. to produce a slurry of finely divided collagen in water, treating the slurry with acid at a pH of 2.5 to 3.7 to swell the collagen, (the ground collagen is suitably swollen by a solution of lactic acid or citric acid), extruding the slurry through an annular die to form a collagen tube, immersing the tube in a coagulating bath containing 0 to 5% by weight of a non-toxic dialdehyde, tanning the collagen tube by contacting the same with a buffered solution of an edible non-toxic dialdehyde at a pH of 4A0 to 7.0, and washing, plasticizing and drying the tube to produce a translucent, non-fibrous, edible product. In another preferred method, a fresh or salt-cured animal hide is treated with a lime-containing solution (suitably a slurry of solid lime, sodium sulfhydrate (NaSH), and dimethylamine sulphate), for a time sufficient to at least partially de-hair the hide, removing the epidermal layer and remaining hair, neutralizing the lime in the hide by treatment with a non-toxic acid capable of forming a soluble calcium salt, at a pH of 2.5 to 6.5, and washing to remove by-product salts, grinding the hide at a temperature less than 20‹ C. to produce a slurry of finely divided collagen in water, treating the slurry with acid at a pH of 2.5 to 3.7 to swell the collagen, (weak acids e.g. oitric acid or lactic acid are suitable), extruding the slurry through an annular die to form a collagen tube, immersing the tube in a coagulating bath, tanning the collagen tube by contacting the same with a solution of glutaraldehyde or condensed smoke containing non-toxic dialdehydes in solution at a pH of 4.0 to 7.0, and washing, plasticizing, and drying the tube to produce a translucent non-fibrous edible product. Collagen casings so made are dried and shirred in conventional manner. Suitably, the coagulating bath consists of a concentrated solution of sodium sulphate or ammonium sulphate. In examples (1) a coagulating bath consisting of 42% ammonium sulphate and 0A5% by weight glutaraldehyde is used; (2) a coagulating bath consisting of 35% ammonium sulphate in water is used; (3) 30% ammonium sulphate; (4) 42% ammonium sulphate; (5) 42% ammonium sulphate; (6) 42% ammonium sulphate; (7) 42% ammonium sulphate and 3 to 10% by weight glutaraldehyde; plasticizing baths specified include 5% glycerol in water. Tanning baths employed include in examples: (1) 5% aqueous glutaraldehyde solution buffered to pH 5.3 with sodium hexametaphosphate; (2) 25% by weight glutaraldehyde solution buffered to pH 5.7 with sodium formate; (3) a concentrated solution of condensed smoke adjusted to a pH of 4.5 (Griffith SF 12 liquid smoke); (4) a solution of 10% by weight glutaraldehyde and 5% by weight " liquid smoke" having a pH of 5.3; (5) a solution of 10% by weight glutaraldehyde buffered to pH 5.9 with sodium formate; (6) a solution of condensed smoke made by passing smoke from sawdust of wood shavings burnt under controlled combustion conditions and passed through water, having a pH of 5.2; (7) a solution of 10% by weight glutaraldehyde and 5% " liquid smoke ", buffered to pH 4.9. The only plasticizing bath disclosed is 5% glycerol in water. In apparatus as shown, the collagen slurry is introduced through inlet conduit 1 into die 2 having an annular die outlet 3 through which casing 4 is extruded. Die 2 has an inner tube 5 extending upwardly within the extruded casing to remove coagulating bath from therewithin. Die 2 is located at the bottom of container 6 which contains coagulating bath 7, bath 7 being circulated through conduit 8 from tube 5 for removal of the coagulating bath from inside the extruded casing. Casing 4, coagulated in bath 7, passes over a series of rollers and is directed through the tanning, washing, and plastiaizing baths shown, to the drying, reeling, and shirring stages.