scholarly journals Dipotassium tetramethyl osmate: a stain for electron microscopy.

1975 ◽  
Vol 23 (2) ◽  
pp. 123-127 ◽  
Author(s):  
C C Hinckley ◽  
J A Murphy

Methanol solutions of dipotassium tetramethyl osmate (DTMO) have been found to be useful as general stains in electron microscopic studies of plant and fungal ultrastructure. The stain solutions are easy to prepare, stable when anhydrous and convenient to use. Although generally similar in staining to lead citrate stains, some elements of cell ultrastructure appear different with dipotassium tetramethyl osmate staining, particularly the outer cell walls of fungi. Indications of specific precipitate-producing reactions in cell storage areas are observed.

1968 ◽  
Vol 39 (1) ◽  
pp. 43-48 ◽  
Author(s):  
Victor J. Matukas ◽  
George A. Krikos

Past work has suggested that protein polysaccharide may play a role in the calcification of cartilage. Recent electron microscopic studies on noncalcified cartilage have indicated that protein polysaccharide in cartilage matrix is represented by granules associated with collagen fibers. The present work has been designed for comparison of the matrix of noncalcified cartilage to that of calcified cartilage, with particular reference to these granules. Small blocks of tibia from 16-day embryos were fixed in cacodylate-buffered glutaraldehyde and postfixed in either phosphate- or Veronal-buffered osmium tetroxide. Special care was taken to maintain the pH above 7.0 at all times. For electron microscopy the tissues were dehydrated, embedded in Epon 812, sectioned, and stained with uranyl acetate or lead citrate. A marked decrease in the size of granules in the matrix of calcified cartilage compared to noncalcified cartilage was noted. Associated with the decrease in the size of granules was a condensation of matrix components and the presence of an amorphous electron-opaque material that was not seen in noncalcified areas. These results are interpreted to represent either a drop in concentration or a change in state of protein polysaccharide with the onset of calcification in cartilage.


Author(s):  
Juith A. Murphy ◽  
C.C. Hinckley

Methonal solutions of dipotassium tetramethyl osmate (DTMO) have ben found to be effective general stains in ultrastructure studies of plant and fungal tissue. The contrast produced by the stain, though broadly similar to that produced by lead citrate, reveals some elements of cell structure not usually seen in the outer cell walls of fungi. DTMO is also without troubling precipitate problems, and in general, less preparatory and washing steps and fewer neccessary precautions. Additionally, region specific precipitates are found in DTMO stained sections of fungi and algae, possibly indicative of chemical reactions between DTMO and reduced cell compenents.


1972 ◽  
Vol 18 (6) ◽  
pp. 534-538
Author(s):  
Mario Werner ◽  
Albert L Jones

Abstract To improve the characterization of electrophoretic lipoprotein subfractions, we developed two new techniques for analyzing lipoproteins after electrophoresis on thin agarose layers. Overlay with antisera exactly localizes specific apoproteins without any distortion caused by antigen diffusion; electron microscopy of eluted fractions determines the varying particle-size distribution. Applied together, these methods can detect individual differences between hyperlipemic samples that are not immediately apparent in the electrophoretic pattern, and should provide valuable new insight into the classification of hyperlipoproteinemias.


1982 ◽  
Vol 202 (3) ◽  
pp. 677-686 ◽  
Author(s):  
F Waechter ◽  
P Bentley ◽  
M Germann ◽  
F Oesch ◽  
W Stäubli

The distribution of rat liver epoxide hydrolase in various subcellular fractions was investigated by immuno-electron-microscopy. Ferritin-linked monospecific anti-(epoxide hydrolase) immunoglobulins bound specifically to the cytoplasmic surfaces of total microsomal preparations and smooth and rough microsomal fractions as well as the nuclear envelope. Specific binding was not observed when the ferritin conjugates were incubated with peroxisomes, lysosomes and mitochondria. The average specific ferritin load of the individual subcellular fractions correlated well with the measured epoxide hydrolase activities. This correlation was observed with fractions prepared from control, phenobarbitone-treated and 2-acetamidofluorene-treated rats.


1997 ◽  
Vol 41 (2) ◽  
pp. 401-409 ◽  
Author(s):  
A Turcotte ◽  
M Simard ◽  
N J Morin ◽  
D Beauchamp ◽  
M G Bergeron

The differential tissue distributions of aztreonam and ceftazidime within fibrin clots infected with Pseudomonas aeruginosa, Enterobacter cloacae, and Serratia marcescens, their efficacies, and the in vivo bacterial morphological changes induced by these drugs were evaluated. Rabbits were given intravenously a single dose of 100 mg of either agents/kg of body weight. In the cores of the clots, the peak levels of both drugs were much lower than those observed in the peripheries and in serum. Aztreonam's half-lives within the peripheries and in the cores of the fibrin clots were up to six times higher than observed in serum, while ceftazidime's half-lives in clots were twice that observed in serum. This resulted in a much greater penetration ratio for aztreonam than for ceftazidime. Both drugs controlled the growth of P. aeruginosa in vivo, but E. cloacae and S. marcescens responded better to ceftazidime. Morphological changes were more abundant in the peripheries than in the cores of the clots. In the control group, P. aeruginosa's morphology in the cores was different than that in the peripheries of the clots. Against P. aeruginosa, aztreonam did induce morphological changes in the cores while ceftazidime did not. Electron microscopic studies revealed that morphological changes associated with aztreonam seemed different than those of ceftazidime. Along with elongation of bacteria, more bow tie and herniated bacteria were observed with aztreonam. Though both agents selectively affect PBP 3, as manifested by elongated bacteria, they induce in the peripheries of the clots thickening, breaks, and detachment in bacterial cell walls, alterations which are generally associated with antibiotics affecting PBP 1a and 1b.


Author(s):  
P.E. Conen ◽  
J.U. Balis ◽  
C.D. Bell

Myogenesis in man was studied using muscle from 19 fetuses of 8 to 16 weeks gestation which were processed with standard osmium-Epon or glutaraldehyde-osmium-Epon schedules and sections were stained in uranyl acetate and/or lead hydroxide. Particular emphasis was given during this study to presence of basement membrane and myofilaments as additional aids in classification of cell types present in developing muscle.Electron microscopy permits accurate identification of fibroblasts and early cells of muscle series and has been used in studies of myogenesis in chick, and rat. Light microscopy definitions for premyoblasts and myoblasts, and for myocytes at the myotube and muscle fiber stages of development are difficult to apply to electron microscopic studies without modification. For example the term myoblast was used differently by Tello, Katznelson and Boyd to designate a cell destined to become muscle but not recognizable as a muscle cell.


Author(s):  
Ralph M. Albrecht ◽  
Scott R. Simmons ◽  
James R. Prudent ◽  
Chris M. Erickson

Colloidal gold, conjugated to a number of biologically active molecules, including ligand and antibody, provides a useful label for light microscopy and electron microscopy. This stems, in part, from its color, density, and regular spherical shape although the ability to make the particles in a number of defined sizes, the ease of conjugation to biological material, and the retention of activity of bound molecules are also important factors.Although nearly all sizes of colloidal gold particles, from 2.0 nm on up, can be identified in transmission or high voltage transmission electron microscopy, it has generally been the larger sized particles, 15 nm and up, that have proved useful for scanning electron microscopic studies. This is due principally to the resolution limits of conventional SEMs and the need to employ backscattered electron imaging, BEI, to unambiguously define the gold labels.


Author(s):  
J.A. Nordquist ◽  
K. Chrysant ◽  
A.K. Mandal

By electron microscopy elastic tissue appear electrolucent in osmium fixed unstained grids as well as grids stained with uranyl acetate and lead citrate (UA + LC). Albert and Fleischer have studied aorta of mice with metalloporphyrins imparting conspicuous electron density to the elastic tissue. We are reporting here a modified method of electron microscopic (EM) study of the elastic tissue using metalloporhyrin, silver tetraphenyl porphyrin sulfonate (STPPS).We have studied the renal arterioles of rats and human in normal and diseased states. Elastic tissue of the aorta from young normal rat served as control for this study. Renal and aortic tissues were fixed in 4 percent glutaraldehyde, post fixed in 1 percent osmium tetroxide and embedded in spurr (blocks). From the blocks of renal tissue, 0.5 μ sections were cut, stained with methylene blue and azure II and studied by light microscopy.


Author(s):  
A. Lupulescu

Previously it has been shown that long-term topical application of 3-methylcholanthrene (MCA) on the rat skin induced basal cell carcinoma. These tumors are very similar to that occurring in humans and they were studied only by light microscopy.1 Transmission electron microscopy (TEM) and Scanning electron microscopy (SEM) can provide more characteristic details for the neoplastic transformation of basal cells, their cytoarchitecture and migration.


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