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At least 19 recordsLinked to original sources
PubMed1993

Analysis of sequence contexts flanking T.G mismatches leads to predictions about reactivity of the mismatched T to osmium tetroxide.

Osmium tetroxide and hydroxylamine are used to detect mutations in DNA and RNA after hybridization of mutant and wild-type DNA. Mismatched T and C bases, respectively, are modified by these reagents and the DNA strand cleaved at the mismatched bases by subsequent treatment with piperidine. This allows detection and location of the mutation. Although most T.G mismatches have been reported to be reactive to osmium tetroxide, some have been reported to be unreactive. The aim of this study was to collect and analyze the reactive and unreactive T.G mismatches. We have collected sequence contexts of all reactive and unreactive T.G mismatches for analysis. This involves 10 unreactive T.G mismatches (plus one T.C) and 19 reactive T.G mismatches. Sequence effects of bases surrounding these mismatches must influence this reactivity. There must be many types of such sequence effects. We postulate that because of the dominance of 5' G bases near the T of unreactive T.G mismatches and the absence of 5' G bases in reactive T.G mismatches that the stacking of the 5' G on the mismatched T is the reason for this lack of reactivity in the majority of the cases studied here.

Base Sequence↗
PubMed2001

Traumatic osmium tetroxide inoculation.

Osmium tetroxide is a highly oxidizing, corrosive compound commonly found in electron microscopy laboratories. Although osmium tetroxide is known to cause rapid damage to organic tissue, its cutaneous effects have not been well studied. We report a case of traumatic inoculation from a broken vial of 4% osmium tetroxide. Electron microscopy and energy dispersive x-ray spectroscopy confirmed the presence of osmium in the tissue specimen. The lesion was treated by simple excision.

Accidents↗
PubMed1995

Osmium tetroxide and ruthenium tetroxide are complementary reagents for the preparation of epidermal samples for transmission electron microscopy.

Ruthenium tetroxide and osmium tetroxide were compared as post-fixatives in the preparation of human epidermis for transmission electron microscopic examination. Both reagents revealed characteristic lamellar granules within the granular layer and extruded lamellar granule contents in the upper granular layer. The transformation of the granule contents into multilamellar sheets at the interface between the granular and cornified layers and the persistence of these sheets through all levels of the stratum corneum were demonstrated only with ruthenium tetroxide fixation. Therefore, the reactivity of osmium tetroxide with isolated epidermal lipids was examined. The failure of osmium tetroxide to reveal membrane structures in the stratum corneum can be explained by its inability to react with many of the lipid components of these membranes, rather than to selective removal of lipids during tissue processing, as was formerly believed. Ruthenium tetroxide, a stronger oxidizing agent than osmium tetroxide, overcomes this problem but has other severe limitations as a post-fixative.

Fixatives↗
PubMed1984

Ultrastructural localization of acetylcholinesterase activity by means of the electron dense precipitate derived from Koelle's cuprous thiocholine iodide by treatment with phosphomolybdic acid and osmium tetroxide.

An osmium resistant, thermostable and electron dense precipitate was obtained from cuprous thiocholine iodide (Koelle's precipitate) by a joint interaction with phosphomolybdic acid and OsO4. No diffusion artifact due to the conversion of the primary precipitate to the secondary precipitate was observed, contrary to that seen after (NH4)2S or K3Fe (CN)6 posttreatment of the cuprous thiocholine iodide. In addition, phosphomolybdic acid and OsO4 provided a counterstain effect on the ultrastructural background. By the present modification, Koelle's histochemical method becomes a useful cytochemical method for ultrastructural localization of acetylcholinesterase activity.

Acetylcholinesterase↗
PubMed1976

The chemical nature of osmium tetroxide fixation and staining of membranes by x-ray photoelectron spectroscopy.

X-ray photoelectron spectroscopy was used to determine the oxidation states of osmium compounds present in erythrocyte ghost preparations and related systems treated with osmium tetroxide. Osmium tetroxide and cholesterol, codeposited at -100 degrees C, began to react at -70 degrees C, and Os(VI) was formed. Similarly, Os(VI) was detected for the known cholesterol-osmate ester prepared and purified chemically. However, osmium tetroxide applied in phosphate buffer (pH 7.2) gave rise to large proportions of Os(IV) and Os(III) species in addition to Os(VI) compounds. Egg phosphatidylcholine likewise produced a mixture of Os(VI), Os(IV), and Os(III), but dipalmitoyl phosphatidylcholine failed to give significant amounts of osmium containing products under identical conditions. Glutaraldehyde gave a mixture of compounds with the same osmium oxidation states when allowed to react with aqueous osmium tetroxide. Unfixed and glutaraldehyde-fixed erythrocyte ghosts also produced mixtures of Ss(VI), Os(IV) and Os(III) under conditions identical to those of normal tissue processing. Additionally, the mixture of adducts initially formed by treatment with osmium tetroxide was further reduced by dehydration of the tissue with ethanol, rpesulting in a final mixture which was 50-60% Os(III). The results support a scheme for the reaction os osmium tetroxide with tissues in which the initial reaction site is the double bonds of unsaturated lipids to form Os(VI) derivatives. Subsequent hydrolysis and further reduction yield complexes of Os(IV) and Os(III). A mixture of these three states is present in membrane specimens during microscopic observation. Os(VI) and Os(IV) could be present as osmate esters and osmium dioxide, respectively; Os(III) could be present as an oxo- or amino complex(es). The photoelectron spectrum of intact erythrocyte ghosts can be synthesized from the spectra of phospholipid and cholesterol only, suggesting the predominance of the reaction with lipids in the fixation process.

Animals↗
PubMed2004

Simultaneous fixation using glutaraldehyde and osmium tetroxide or potassium ferricyanide-reduced osmium for the preservation of monogenean flatworms: an assessment for Merizocotyle icopae.

Simultaneous fixation was investigated for a marine organism: the monogenean flatworm ectoparasite Merizocotyle icopae. Four protocols for primary fixation were compared: 3% glutaraldehyde alone in 0.1M cacodylate buffer for a minimum of 2 hours; 1% glutaraldehyde in combination with 1% osmium tetroxide, both in 0.1M cacodylate buffer, until tissues darkened (5-20 minutes); 1% glutaraldehyde in 0.1M cacodylate buffer in combination with 0.5% potassium ferricyanide-reduced osmium until tissues darkened (5-20 minutes); 1% glutaraldehyde in combination with 1% osmium tetroxide, both in 0.1M cacodylate buffer, for 30 minutes. The study confirms that the standard method for transmission electron microscopic fixation (first listed protocol) routinely applied to platyhelminths is optimal for ultrastructural preservation, but some simultaneous fixation methods (second and third listed protocols) are acceptable when rapid immobilization is needed. Scanning electron microscopic preparations may be improved using simultaneous primary fixation.

Animals↗
PubMed1988

Increase in the calcium content of cardiac tissue after postfixation with osmium tetroxide.

The concentration of osmium has been measured by destructive chemical analysis in glutaraldehyde fixed heart tissue postfixed with osmium tetroxide and embedded in epoxy resin. After such treatment, the mean atomic number of the specimen (Z) is close to 10, which permits a quantitative analysis of calcium (Ca) by the continuum method, using Z2/A as a correcting factor (A: atomic weight). Wavelength-dispersive X-ray microanalysis has been used to determine the Ca concentration of frog cardiac tissue fixed in glutaraldehyde and embedded in resin. These measurements have been repeated on tissue postfixed in osmium tetroxide; contrary to expectations, the apparent Ca concentration is much higher in osmium treated than in nontreated tissue. However, this result is observed with OsO4 solutions prepared in glass, not with solutions prepared in plastic. It is shown by energy dispersive X-ray analysis of droplets that OsO4 solutions prepared in glass contain large amounts of calcium, potassium and silicon. Care must be taken in preparing OsO4 fixatives when the fixed tissues are to be subjected to X-ray microanalysis of such elements as Ca or Si.

Animals↗
PubMed1982

Imidazole-buffered osmium tetroxide: an excellent stain for visualization of lipids in transmission electron microscopy.

The usefulness of imidazole-buffered osmium tetroxide as a stain for lipids in transmission electron microscopy has been investigated. Rat liver and other tissues were fixed by perfusion with glutaraldehyde and post-fixed with osmium-imidazole and the appearance of lipid droplets was compared with that after post-fixation in unbuffered aqueous osmium tetroxide or an osmium solution buffered otherwise. Prominent electron-opaque staining of lipid droplets and of lipoprotein particles was noted after post-fixation with 2% osmium-imidazole, pH 7.5, for 30 min. The lipid droplets appeared well circumscribed with no evidence of diffusion. In contrast, the intensity of staining was much less and there was some diffusion around lipid droplets in material post-fixed in aqueous or cacodylate-buffered osmium tetroxide. Spot tests on filter paper revealed that unsaturated fatty acids, especially linolenic and linoleic acids reacted more intensely with osmium-imidazole than with aqueous osmium tetroxide. These findings demonstrate that osmium-imidazole provides an excellent stain for lipids in transmission electron microscopy and that most probably it stains lipids with unsaturated fatty acids.

Adipose Tissue, Brown↗
PubMed1988

Probing of DNA polymorphic structure in the cell with osmium tetroxide.

It is shown that osmium tetroxide, 2,2'-bipyridine can be applied as a probe of DNA structure in a bacterial cell. Using this probe we demonstrate (a) presence of structural distortions at the junctions between the right-handed B and left-handed Z DNA in a recombinant plasmid pRW751 and (b) unusual structure of the d(A-T)16 insert in pAT32 plasmid in E. coli cells and in in vitro.

DNA Restriction Enzymes↗
PubMed1998

Investigation of the ultrastructurel of different cells in lymph nodules by using zinc iodide osmium tetroxide technique.

The zinc iodide -osmium tetroxide (ZIO) fixation/staining method was applied for neurocytological study and also to examine several other tissue samples including human blood and bone marrow on nerve endings in the median eminence, epidermal langerhans cells of lymphoid tissue. Although precise specificity can not be attributed to the staining reaction. Interesting staining patterns for different cell types in lymph node were observed by one of the ZIO staining solutions. The significance of ZIO positivity is briefly discussed.

Animals↗
PubMed1998

Light and electron microscopic examination of exocrine pancreas using zinc iodide-osmium tetroxide technique.

Zinc iodide-osmium tetroxide (ZIO) fixation/staining technique is a metallophilic technique which has been used for the examination of various tissues and cell types. We examined the ZIO (+) cell types in rat exocrine pancreas to obtain further evidence for the significance of the reaction. Among mostly non-reactive pancreatic acinar cells there were ZIO (+) acinar cells of varying staining intensity. Zymogenic granules and centroacinar cells were completely non-reactive. Our electron microscopic findings support the view that the reactivity of the technique used is cell specific but not cell type or organelle specific.

Animals↗
PubMed2004

Voltammetric behavior of DNA modified with osmium tetroxide 2,2'-bipyridine at mercury electrodes.

Osmium tetroxide complexes with nitrogen ligands (L) are probes of DNA structure and electroactive labels of DNA. Here adducts of single-stranded (ss) DNA with osmium tetroxide 2,2'-bipyridine (DNA-Os,bipy) were studied by cyclic voltammetry for the first time. It was found that at neutral pH DNA-Os,bipy produces three redox couples in the potential range between 0 and -1 V (peaks I-III) and a cathodic peak at about -1.3 V (peak IV). The latter peak decreased with increasing scan rate, and peaks arising from the forward and reverse scans exhibited the same direction, suggesting catalytic nature of the electrode process. We concluded that this peak corresponds to the known differential pulse voltammetric (polarographic) peak of DNA-Os,L adducts for which catalytic hydrogen evolution is responsible. In contrast, currents of cathodic peaks II and III increased almost linearly with increasing scan rate, suggesting involvement of adsorption in the electrode processes. Adsorptive stripping square-wave voltammetry was used to analyze the DNA-Os,bipy at low concentrations. It was shown that at neutral pH, peak III can offer sensitivity in the ppb range, which is only little lower than that reached by catalytic peak IV. The latter peak is, however, superior in sensitivity at acid pH values.

2,2'-Dipyridyl↗
PubMed1967

Oxidation of pyrimidine nucleosides and nucleotides by osmium tetroxide.

1. Pyrimidine nucleosides such as thymidine, uridine or cytidine are oxidized readily at 0 degrees by osmium tetroxide in ammonium chloride buffer. There is virtually no oxidation in bicarbonate buffer of similar pH. Oxidation of 1-methyluracil yields 5,6-dihydro-4,5,6-trihydroxy-1-methyl-2-pyrimidone. 2. Osmium tetroxide and ammonia react reversibly in aqueous solution to form a yellow 1:1 complex, probably OsO(3)NH. A second molecule of ammonia must be involved in the oxidation of UMP since the rate of this reaction is approximately proportional to the square of the concentration of unprotonated ammonia. 3. 4-Thiouridine reacts with osmium tetroxide much more rapidly than does uridine. The changes of absorption spectra are different in sodium bicarbonate buffer and in ammonium chloride buffer. They occur faster in the latter buffer and, under suitable conditions, cytidine is a major product. 4. Polyuridylic acid is oxidized readily by ammoniacal osmium tetroxide, but its oxidation is inhibited by polyadenylic acid. Pyrimidines of yeast amino acid-transfer RNA are oxidized more slowly than the corresponding mononucleosides, especially the thymine residues. Appreciable oxidation can occur without change of sedimentation coefficient.

Chromatography↗
PubMed1994

New applications for the zinc iodide-osmium tetroxide technique.

The zinc iodide-osmium tetroxide (ZIO) fixation/staining method was applied for neurocytological studies and also to examine several other tissue samples including epidermal Langerhans cells, blood and bone marrow cells and lymphoid tissue. Although precise specificity cannot be attributed to the staining reaction, interesting staining patterns for different cell types were observed by using one of the ZIO staining solutions. The significance of ZIO positivity is briefly discussed.

Animals↗
PubMed1983

Tannic acid- and thiocarbohydrazide-mediated osmium tetroxide binding in the preparation of human leucocytes for SEM observation.

Normal human leucocytes, successively treated with glutaraldehyde-tannic acid-osmium tetroxide-thiocarbohydrazide-osmium tetroxide-thiocarbohydrazide-osmium tetroxide, were prepared for scanning electron microscopy observation. These cells produced well-contrasted, non-charging scanning images compatible with metal-evaporated material. Further, the mononuclear and polymorphonuclear cells resisted shrinkage during dehydration and critical point drying, thus allowing much improved images at high magnification than those covered with evaporated metal. In all cases at least a second thiocarbohydrazide-osmium tetroxide treatment could not be avoided.

Fixatives↗
PubMed1981

Comparative demonstration of pulmonary fat emboli by "en bloc" osmium tetroxide and oil red O methods.

The presence and localization of fat in human lung tissue was evaluated by the "en bloc" staining procedure with osmium tetroxide performed with formalin fixed tissue with subsequent paraffin sectioning and with the oil red O technique performed with frozen sections. Fifty-one autopsy cases were divided into three clinical groups: group A, consisting of 17 patients with severe skeletal trauma; group B, consisting of 14 patients with minor skeletal trauma; and group C, consisting of 20 control patients without trauma. Adjacent sections of lung were selected from all cases, stained with the "en bloc" osmium tetroxide and oil red O methods, coded, and examined under the light microscope without knowledge of the clinical grouping, Stainable fat was graded on a 1 to 4+ scale, and attention was given to histologic localization in tissue sections. The "en bloc" osmium tetroxide technique revealed greater amounts of stainable lipid in clinical groups A and B and was most effective in demonstrating lipid when present in small quantities (group B). Since the method permits the employment of paraffin sections, evaluation of fine histologic detail is an advantage that is not always obtained in frozen sections. It is concluded that the "en bloc" osmium tetroxide technique is superior to the oil red O technique in terms of visualization and histologic localization of minute amounts of lipid in lung tissue.

Adipose Tissue↗
PubMed1995

Complex of osmium tetroxide with 1,10-phenanthroline binds covalently to double-stranded DNA.

Complex of osmium tetroxide with 1,10-phenanthroline (Os,phen) reacts with double-stranded B-DNA in contrast to osmium tetroxide, pyridine and other osmium structural probes which show a strong preference for single-stranded DNA (ssDNA) (Palecek, E. in Abelson, J.N., and Simon, M.I. (eds), Lilley, D.M.J., and Dahlberg, J.E., (volume eds.), Methods in Enzymology, Vol. 212, DNA Structures, part B., Academic Press, 139-155 (1992)). Modification of negatively supercoiled DNA (scDNA) with Os,phen changes the DNA electrophoretic mobility inducing the DNA relaxation at lower degrees of modification followed by formation of positive supercoils at higher modification extents. Electrophoretic mobility of the Os,phen-modified DNA fragments in agarose gel is almost unchanged while a strong retardation of the same fragments is observed in polyacrylamide gels. Os,phen-modified DNA is hypersensitive to nuclease S1. Cleavage of this DNA by restriction enzymes is selectively inhibited showing a preference of Os,phen for TA and AT dinucleotide steps. DNA modification by Os,phen is inhibited by low and moderate concentrations of MgCl2. The covalent binding of Os,phen to double-stranded DNA (dsDNA) is preceded by noncovalent interactions (probably intercalation) inducing DNA structural changes; the shape of the Os,phen-modified DNA molecule appears to be severely deformed.

Base Composition↗