Osmium tetroxide solution

2006 ◽  
Vol 2006 (1) ◽  
pp. pdb.rec447
2000 ◽  
Vol 6 (S2) ◽  
pp. 844-845
Author(s):  
O.J. Castejón ◽  
R. P. Apkarian ◽  
H. V. Castejón

Samples of albino mice cerebellar cortex were processed by the cryofracture method for scanning electron microscopy and examined with the field emission scanning electron microscope (FESEM). Albino mouse cerebellar cortex was excised, cut into 1-2 mm slices and inmersed in 4% glutaraldehyde in O. l M phosphate buffer solution, pH 7.4, for 24h at 4°C; and postfixed for 1 h in a similarly buffered 1% osmium tetroxide solution. Specimens were dehydrated in a graded serie of ethanol (30, 50, 70, 80, 90 2x100%) prior to wrapping individual tissue pieces in preformed absolute ethanol filled parafilm cryofracture packets. Rapid freezing of packets was performed by plunging into LN2. First, the packet was transferred from the LN2 storage vessel with LNT chilled forceps in order to avoid themial damage. Secondly, the cooled fracture blade was removed from the LN2, the packet was orientated under the blade, and immediately struck with a heavy tool.


Author(s):  
Tatsuya Matsuo ◽  
Mikio Suzuki

Recently, very thick section of biological specimens have been observed by using super high voltage electron microscope to obtain a three dimensional model of a specimen.However, the thicker the section, the more difficult the penetration of staining solution into the depth of thick section. So, it is doutful whether the stain solution fully penetrated into the thick section. This paper reports an observation of various thick unstained sections using scanning transmitted electron microscope(STEM) and conventional electron microscope(CEM).The observed specimen is mice liver cells. The tissue was fixed in cold 1% phosphate-buffered osmium tetroxide solution for 2 hours. After fixation, the tissue was dehydrated in a graded series of ethanols and then embedded in Epon 812. The 500 Å, 5,000 Å and 1μ thick sections were cut, not stained with uranyle and lead, and then examined with STEM and GEM operated at accelerating voltage 80 kV.


1940 ◽  
Vol s2-81 (324) ◽  
pp. 595-617
Author(s):  
J. BRONTÉ GATENBY ◽  
J. D. SMYTH

1. In Chilomonas paramecium the contractile vacuole is surrounded by a cortical substance (Golgi apparatus) which has the power of reducing osmium tetroxide solution and thus impregnating black (Nassonow). 2. This cortex blackens thus in over 99 per cent, of individuals in a culture which has not been dividing. In a culture in which the individuals have been rapidly dividing, the percentage of unimpregnated contractile vacnoles increases considerably, up to about 5 per cent. 3. During division of Chilomonas in about 70 per cent. of cases the osmiophile substance is very equally divided between the daughter cells. The dividing cortex comes away from the contractile vacuole, which eventually collapses, new contractile vacuoles arising in the site of the divided osmiophile material. In about 25 per cent, of division stages osmication of the cortex fails to a greater or lesser degree. There is always a very distinct tendency for this failure to take place even in the best of preparations. 4. In some cases (about 3 per cent.), during division, the entire contractile vacuole and its cortex goes over whole to one individual. A new vaeuole, apparently without cortex, arises spontaneously in the other individual. It is unlikely that all of these cases are due to failure of impregnation in one of the individuals, though this possibility cannot be roled out completely. 5. The behaviour of the original contractile vacuole cavity before separation of the daughter cells is as follows. The lipoid, having partially retreated from the vacuole, becomes separated into two parts, and the centrally placed vacuole disappears (figs. 4 and 6, Pl. 36; figs. 10 and 15, Pl. 37). New vacuoles appear in the site of the lipoid bodies in each daughter cell (fig. 5, Pl. 36). 6. Two ellipsoidal accessory bodies or pyrenoids lie on a level with the vestibule. In older cultures the two bodies are often exactly the same size and colour (corrosive osmic followed by neutral red or haematoxylin), but in rapidly dividing cultures, one body may be of normal size, whereas the other may be absent or much smaller. During cell division, one body is carried across to each daughter. No exception to this was ever found. 7. Identification of the smaller Peranemidae is in a confused state. Probably several species, and possibly even genera, have been described by various authors as Scytomonas (Copromonas) subtilis.


1975 ◽  
Vol 23 (8) ◽  
pp. 575-598 ◽  
Author(s):  
J A Simson ◽  
S S Spicer

Fixation of rat parotid with an unbuffered osmium tetroxide solution containing nearly saturated potassium (pyro)antimonate resulted in abundant deposition of cation-antimonate precipatates in acinar cells. Altering the antimonate concentration, including buffers or chelators in the solution or changing the primary fixative resulted in an altered intensity and distribution of the precipitates formed in the tissue, apparently reflecting a degree of selectivity in ion localization. Decreasing the concentration of pyroantimonate to about half-saturation preserved predominantly the less soluble antimonate salts (e.g., Na+, Ca++) and resulted in preferential retention of deposits along the plasmalemma and in mitochondrial "dense bodies," with loss of most cytoplasmic and nuclear precipitates. A similar pattern was seen if fixation with the high concentration antimonate-osmium procedure was followed by a prolonged rinse. Adding phosphate or collidine buffers markedly decreased precipitates in the nuclei and on granular reticulum as well. Phosphate buffer or ehtyleneglycoltetraacetate inhibited in vitro precipitation of calcium and sodium and decreased or abolished plasmalemmal deposits. Glutaraldehyde fixation, either in the presence of antimonate or prior to antimonate-containing osmium tetroxide, abolished heterochromatin deposits. Mitochondrial dense bodies were of two types, one containing precipitate and the other inherently osmiophilic. The latter were also observed in pyrophosphate-osmium controls. Results from in vitro titrations of cations with the various antimonate methods and from neutron activation analyses of fixed tissues supported conclusions drawn from fine structural distribution patterns and were interpreted as follows. In rat parotid acinar cells, deposits in heterochromatin and on granular reticulum probably arose from precipitation in sites of high K+ and H+ as well as--NH3+-rich histones. Plasmalemmal antimonate deposits demonstrated sites of sodium and/or calcium accumulation. Some mitochondrial dense bodies contained Ca++ whereas others were inherently osmiophilic. Large, extracellular deposits were probably predominantly sodium precipitates.


1969 ◽  
Vol 17 (2) ◽  
pp. 102-106 ◽  
Author(s):  
BERNARD P. LANE ◽  
EUGENE MARTIN

Electron microscopy of Epon-embedded mouse vas deferens eipthelium treated with buffered potassium pyroantimonate-osmium tetroxide solution revealed precipitates in the lamina propria and along the apical plasma membrane. Electron microprobe elemental analysis of adjacent sections demonstrated that the deposits contained sodium and antimony. Other cations noted to precipitate pyroantimonate in vitro were not present in large amounts compared to controls, and were randomly distributed.


1961 ◽  
Vol 11 (2) ◽  
pp. 273-296 ◽  
Author(s):  
H. E. Huxley ◽  
G. Zubay

Oriented fibres of extracted nucleohistone were employed as test material in a study of satisfactory fixation, embedding, and staining methods for structures containing a high proportion of nucleic acid. Fixation in buffered osmium tetroxide solution at pH 6, containing 10-2 M Ca++, and embedding in Araldite enabled sections of the fibres to be cut in which the orientation was well preserved. These could be strongly stained in 2 per cent aqueous uranyl acetate, and showed considerable fine structure. Certain regions in the nuclei of whole thymus tissue could also be strongly stained by the same procedure, and were identical with the regions stained by the Feulgen procedure in adjacent sections. Moreover, purified DNA was found to take up almost its own dry weight of uranyl acetate from 2 per cent aqueous solution. Strongest staining of whole tissue was obtained with very short fixation times-5 minutes or so at 0°C. Particularly intense staining was obtained when such tissue stained in uranyl acetate was further stained with lead hydroxide. Although the patterns of staining by lead hydroxide alone and by uranyl acetate were similar in tissues fixed for longer times (½ hour to 2 hours, at 0°C or 20°C), in briefly fixed material the DNA-containing regions appeared relatively unstained by lead hydroxide alone, whilst often there was appreciable staining of RNA-containing structures. Observations on the staining of some viruses by similar techniques are also described.


1972 ◽  
Vol 20 (7) ◽  
pp. 518-526 ◽  
Author(s):  
S. S. SPICER ◽  
A. A. SWANSON

Elements retained in cervical lymph nodes, isolated hepatic nuclei and salt-impregnated gels by fixation with antimonate- or pyrophosphate-containing and other osmium tetroxide solutions were assayed by nuclear activation analysis or by atomic absorption spectrophotometry. Salts preserved by the antimonate-osmium tetroxide fixative in lymph nodes, isolated nuclei and a KCl-enriched gel consisted almost entirely of potassium antimonate. The K+ in the precipitates in these specimens appeared to derive partially from that in the fixative solution and partially from that in the specimen. Salts preserved by the antimonate-osmium tetroxide fixative in an NaCl-supplemented gel consisted partly of potassium antimonate derived from the fixative as in unsupplemented gels and partly of sodium antimonate. The Na+ precipitated in this gel amounted to less than one-half that originally present. In comparison the pyrophosphate-osmium tetroxide solution retained higher levels of K+ in lymph nodes, nuclei and the KCl gel, but the potassium pyrophosphate was not evident as electron-opaque precipitates. The latter fixative was less effective in preserving Na+ in the NaCl gel. The pyrophosphate-containing fixative, which was about twice as efficient as the antimonate-containing solution in retaining the divalent cations, preserved 70% of the Mg++ and 100% of the Ca++ added to gels.


Author(s):  
T. Ohkura ◽  
H. Iwatsuki ◽  
T. Watanabe

As described in a previous report, Alcian blue provides the selective contrast enhancement of interfibrillar substances of the mesostroma of the chick embryo cornea. This finding was also verified by the electron probe microanalysis of adjacent sections. In the present study the nature of interfibrillar substances was ultrahistochemically examined by a modified Seligman's method for the demonstration of some oxidizable glycols.Strips of the 4th day chick embryo.cornea were fixed in ice-cold 2.5% glutaraldehyde solution (pH 7.4). Tissue specimens treated with 1% aqueous periodic acid or 0.2% hyaluronidase solution of pH 6.0 for 15 min were transferred into 1% thiocarbohydrazide, followed by a rinse in 1% osmium tetroxide solution (60°C, 50 min).


1969 ◽  
Vol 17 (12) ◽  
pp. 781-792 ◽  
Author(s):  
S. S. SPICER ◽  
W. B. GREENE ◽  
J. H. HARDIN

For selective ultrastructural localization of acid mucosubstance in rabbit and human platelets and megakaryocytes, bone marrow and buffy coat specimens were fixed with formalin, glutaraldehyde or osmium tetroxide, sectioned at 40 µ and stained with the Rinehart-Abul-Haj solution of dialyzed iron. In specimens from both rabbit and man, dialyzed iron staining was observed within nucleoids of the cytoplasmic granules (α-granulomeres) of platelets and megakaryocytes, on the outer surface of the plasma membranes of platelets and megakaryocytes and on the luminal surface of demarcation membranes of megakaryocytes. These results were obtained following any of the three fixation procedures, except when nucleoids failed to stain after glutaraldehyde fixation. For ultrastructural localization of pyroantimonate-precipitable cation, bone marrow and buffy coat specimens were fixed in Komnick's solution of potassium pyroantimonate and osmium tetroxide. In specimens from both species, antimonate deposits were localized within the dense bodies (5-hydroxytryptamine organelles) of platelets and within nucleoids of cytoplasmic granules of platelets and megakaryocytes. The dense bodies were well preserved in platelets fixed in a pyrophosphate-osmium tetroxide solution but were poorly, if at all, preserved by osmium tetroxide solutions containing other buffers.


1966 ◽  
Vol 35 (3) ◽  
pp. 271-279 ◽  
Author(s):  
D. G. MONTEMURRO

SUMMARY The ventral hypothalamus and the pituitary gland of the rat were examined with the light microscope in sections fixed in an osmium tetroxide solution without further staining. The histology of the neurohypophysis could be observed with remarkable clarity. Herring bodies and axons of the supra-optico-neurohypophysial tract show as grey homogeneous structures. Pituicytes appear as round or variously shaped cells whose cytoplasm is crowded with osmiophilic granules. The granules vary in size from 0·5 to about 2·5 μ, and present either as solid or as ring-shaped black structures. They stained neither with aldehyde-thionine, nor with chrome-alum haematoxylin, nor with the PA-Schiff techniques but stained with Sudan black B. They were found in profusion in the median eminence of the hypothalamus and the infundibular stem in an apparent extracellular location surrounding fibre bundles and capillaries of the hypophysial portal plexus. Two types of cells were identifiable in the supraoptic nucleus of the hypothalamus. Smaller, angular neurones stained densely with osmium and with aldehyde-thionine and larger, more rounded neurones stained only lightly with osmium and did not stain with aldehyde-thionine. A peculiar laminated arrangement of structures in the infundibular recess in the region of the arcuate nucleus was noted. Several distinct types of cells in the adenohypophysis were identifiable on morphological grounds: thyrotrophic basophils, gonadotrophic basophils, acidophils and chromophobes. The possibility of a neuroendocrine role of the pituicytes is discussed.


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