scholarly journals CRITIQUE ON THE K-PYROANTIMONATE METHOD FOR SEMIQUANTITATIVE ESTIMATION OF CATIONS IN CONJUNCTION WITH ELECTRON MICROSCOPY

1972 ◽  
Vol 20 (1) ◽  
pp. 65-78 ◽  
Author(s):  
RICHARD L. KLEIN ◽  
SHYUE-SHONG YEN ◽  
ÅSA THURESON-KLEIN
Keyword(s):  
Electron Microscopy ◽  
Divalent Cations ◽  
Test Tube ◽  
Tetraacetic Acid ◽  
Test Conditions ◽  
Solubility Products ◽  
Controlled Preparation ◽  
Preparation Techniques ◽  
Potassium Pyroantimonate

The histochemical method employing potassium pyroantimonate in conjunction with electron microscopy has been investigated using carefully controlled preparation techniques and very sensitive atomic absorption analysis of cations. A critique on the reliability and limitations of the method based on test tube and in vitro experiments is given. The method is sensitive to Ca++, Mg++ and Na+ at the 10–6, 10–5 and <10–2 M levels, respectively. Under defined conditions a linear ~l:l ratio of cation present to cation precipitated occurs above these levels. Approximate solubility products have been estimated. Under the test conditions, K+ does not precipitate as a pyroantimonate salt, and neither K+ nor OsO4 influences cation precipitaton at physiologic concentrations. Unbuffered, Tris-HCl-buffered and weakly buffered NaHCO3 media at pH 7.2-7.8 give statistically similar results with Na+ precipitation. The pyroantimonate ion can compete with chelators, ethylenedinitrilotetraacetic acid and ethylene glycol bis-N, N'-tetraacetic acid, for divalent cations when employed simultaneously. These chelators effectively remove Ca++ but not Mg++ from embryonic myocardium, and their effects on Na+ and K+ balance are not marked if employed for relatively short periods. Electron micrographic examples of cation precipitates are given in support of certain findings. A brief discussion of the significance of pyroantimonate grain size, the discrepancy between the ratio of intra- and extracellular precipitates and guidelines for the use of the method are included.

Reversible Changes of Chromosome Structure upon Different Concentrations of Divalent Cations

2019 ◽  
Vol 25 (3) ◽  
pp. 817-821 ◽  
Author(s):  
Astari Dwiranti ◽  
Hideaki Takata ◽  
Kiichi Fukui
Keyword(s):  
Electron Microscopy ◽  
Chromosome Structure ◽  
Ethylenediaminetetraacetic Acid ◽  
Divalent Cations ◽  
Chromosome Organization ◽  
Human Chromosomes ◽  
Tetraacetic Acid ◽  
Structural Details ◽  
Scanning Electron

AbstractThe structural details of chromosomes have been of interest to researchers for many years, but how the metaphase chromosome is constructed remains unsolved. Divalent cations have been suggested to be required for the organization of chromosomes. However, detailed information about the role of these cations in chromosome organization is still limited. In the current study, we investigated the effects of Ca2+ and Mg2+ depletion and the reversibility upon re-addition of one of the two ions. Human chromosomes were treated with different concentrations of Ca2+and Mg2+. Depletion of Ca2+ and both Ca2+ and Mg2+ were carried out using 1, 2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid and ethylenediaminetetraacetic acid (EDTA), respectively. Chromosome structure was examined by fluorescence microscopy and scanning electron microscopy. The results indicated that chromosome structures after treatment with a buffer without Mg2+, after Ca2+ depletion, as well as after depletion of both Mg2+, and Ca2+, yielded fewer compact structures with fibrous chromatin than those without cation depletion. Interestingly, the chromatin of EDTA-treated chromosomes reversed to their original granular diameters after re-addition of either Mg2+ or Ca2+ only. These findings signify the importance of divalent cations on the chromosome structure and suggest the interchangeable role of Ca2+ and Mg2+.

scholarly journals ELECTRON PROBE ANALYSIS OF CATIONIC SPECIES IN PYROANTIMONATE PRECIPITATES IN EPON-EMBEDDED TISSUE

1969 ◽  
Vol 17 (2) ◽  
pp. 102-106 ◽  
Author(s):  
BERNARD P. LANE ◽  
EUGENE MARTIN
Keyword(s):  
Electron Microscopy ◽  
Electron Probe ◽  
Osmium Tetroxide ◽  
Vas Deferens ◽  
Apical Plasma Membrane ◽  
Mouse Vas Deferens ◽  
Electron Probe Analysis ◽  
Osmium Tetroxide Solution ◽  
Potassium Pyroantimonate

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.

Characterization of sea urchin primary mesenchyme cells and spicules during biomineralization in vitro

Development ◽  
1987 ◽  
Vol 101 (2) ◽  
pp. 297-312 ◽  
Author(s):  
G.L. Decker ◽  
J.B. Morrill ◽  
W.J. Lennarz
Keyword(s):  
Electron Microscopy ◽  
Sea Urchin ◽  
Secretory Pathway ◽  
Divalent Cations ◽  
Ruthenium Red ◽  
Coated Pits ◽  
Residual Matrix ◽  
Mesenchyme Cells ◽  
Primary Mesenchyme Cells

An in vitro culture system for primary mesenchyme cells of the sea urchin embryo has been used to study the cellular characteristics of skeletal spicule formation. As judged initially by light microscopy, these cells attached to plastic substrata, migrated and fused to form syncytia in which mineral deposits accumulated in the cell bodies and in specialized filopodial templates. Subsequent examination by scanning electron microscopy revealed that the cell bodies and the filopodia and lamellipodia formed spatial associations similar to those seen in the embryo and indicated that the spicule was surrounded by a membrane-limited sheath derived by fusion of the filopodia. The spicules were dissolved from living or fixed cells by a chelator of divalent cations or by lowering the pH of the medium. However, granular deposits found in the cell bodies appeared relatively refractory to such treatments, indicating that they were inaccessible to agents that dissolved the spicules. Use of rapid freezing and an anhydrous fixative to preserve the syncytia for transmission electron microscopy and X-ray microprobe analysis, indicated that electron-dense deposits in the cell bodies contain elements (Ca, Mg and S) common to the spicule. Examination of the spicule cavity after dissolution of the spicule mineral revealed openings in the filopodia-derived sheath, coated pits within the limiting membrane and a residual matrix that stained with ruthenium red. Concanavalin A—gold applied exogenously entered the spicule cavity and bound to matrix glycoproteins. Based on these observations, we conclude that components of the spicule initially are sequestered intracellularly and that spicule elongation occurs in an extracellular cavity. Ca2+ and associated glycoconjugates may be routed in this cavity via a secretory pathway.

Calcium accumulation in vacuoles of Physarum polycephalum following starvation

1980 ◽  
Vol 44 (1) ◽  
pp. 75-85
Author(s):  
R. Kuroda ◽  
H. Kuroda
Keyword(s):  
Electron Microscopy ◽  
Physarum Polycephalum ◽  
Microprobe Analysis ◽  
Soluble Fraction ◽  
Tetraacetic Acid ◽  
Fresh Weight ◽  
X Ray ◽  
Plasmodium Of Physarum Polycephalum ◽  
Potassium Pyroantimonate ◽  
Extracellular Slime

The plasmodium of Physarum polycephalum contained 15.3 mmol Ca/kg fresh weight of sample, 11.8 mmol Mg/kg, 24.5 mmol K/kg and 1.4 mmol Na/kg. When the plasmodium was starved of food, the Ca content increased gradually up to 71.9 mmol/kg during 5 days of starvation. The concentration of other elements changed only slightly. The endoplasm contained 23.0 mmol Ca/kg, 12.6 mmol Mg/kg, 26.6 mmol K/kg and 1.7 mmol Na/kg, but these contents changed only slightly during starvation. The Ca, Mg, K and Na contents of the slime and the soluble fraction were also determined. In order to clarify where the accumulated Ca was localized, Ca in the plasmodium was precipitated with potassium pyroantimonate and examined by electron microscopy. In the starved plasmodium, the vacuoles which contained the electron-opaque precipitates and were located in the ectoplasm increased in number, compared with the unstarved plasmodium. At the same time the large electron-opaque granules in the extracellular slime increased in number. The electron-opaque precipitates were identified as Ca pyroantimonate by its susceptibility to removal by chelation with ethyleneglycol bis (beta-aminoethyl ether) N, N, N', N'-tetraacetic acid (EGTA) and X-ray microprobe analysis.

Sem Studies of Co-Cr-Mo Surgical Implant Alloy Corrosion Behavior

1982 ◽  
Vol 40 ◽  
pp. 520-521
Author(s):  
Ann Chidester Van Orden ◽  
John L. Chidester ◽  
Anna C. Fraker ◽  
Pei Sung
Keyword(s):  
Electron Microscopy ◽  
Corrosion Behavior ◽  
Anodic Polarization ◽  
Saline Solution ◽  
Saturated Calomel Electrode ◽  
Physiological Saline ◽  
X Ray ◽  
Physiological Saline Solution ◽  
Scanning Electron

The influence of small variations in the composition on the corrosion behavior of Co-Cr-Mo alloys has been studied using scanning electron microscopy (SEM), energy dispersive x-ray analysis (EDX), and electrochemical measurements. SEM and EDX data were correlated with data from in vitro corrosion measurements involving repassivation and also potentiostatic anodic polarization measurements. Specimens studied included the four alloys shown in Table 1. Corrosion tests were conducted in Hanks' physiological saline solution which has a pH of 7.4 and was held at a temperature of 37°C. Specimens were mechanically polished to a surface finish with 0.05 µm A1203, then exposed to the solution and anodically polarized at a rate of 0.006 v/min. All voltages were measured vs. the saturated calomel electrode (s.c.e.).. Specimens had breakdown potentials near 0.47V vs. s.c.e.

Maytansine-Induced Mitotic Arrest in Human Lymphoblasts

1977 ◽  
Vol 35 ◽  
pp. 460-461
Author(s):  
Tai-Te Chao ◽  
John Sullivan ◽  
Awtar Krishan
Keyword(s):  
Electron Microscopy ◽  
Cell Cycle ◽  
Transmission Electron Microscopy ◽  
Tubulin Polymerization ◽  
Vinca Alkaloids ◽  
Antimitotic Activity ◽  
Transmission Electron ◽  
Flow Microfluorometry ◽  
Brain Tubulin

Maytansine, a novel ansa macrolide (1), has potent anti-tumor and antimitotic activity (2, 3). It blocks cell cycle traverse in mitosis with resultant accumulation of metaphase cells (4). Inhibition of brain tubulin polymerization in vitro by maytansine has also been reported (3). The C-mitotic effect of this drug is similar to that of the well known Vinca- alkaloids, vinblastine and vincristine. This study was carried out to examine the effects of maytansine on the cell cycle traverse and the fine struc- I ture of human lymphoblasts.Log-phase cultures of CCRF-CEM human lymphoblasts were exposed to maytansine concentrations from 10-6 M to 10-10 M for 18 hrs. Aliquots of cells were removed for cell cycle analysis by flow microfluorometry (FMF) (5) and also processed for transmission electron microscopy (TEM). FMF analysis of cells treated with 10-8 M maytansine showed a reduction in the number of G1 cells and a corresponding build-up of cells with G2/M DNA content.

Some Structural Features of Metaphase Chromosomes of Mice and Men

1967 ◽  
Vol 25 ◽  
pp. 190-191
Author(s):  
Godfrey C. Hoskins ◽  
Betty B. Hoskins
Keyword(s):  
Electron Microscopy ◽  
Electron Micrographs ◽  
Structural Features ◽  
Metaphase Chromosomes ◽  
The Future ◽  
Of Mice And Men ◽  
Mouse Cells ◽  
Human And Mouse

Metaphase chromosomes from human and mouse cells in vitro are isolated by micrurgy, fixed, and placed on grids for electron microscopy. Interpretations of electron micrographs by current methods indicate the following structural features.Chromosomal spindle fibrils about 200Å thick form fascicles about 600Å thick, wrapped by dense spiraling fibrils (DSF) less than 100Å thick as they near the kinomere. Such a fascicle joins the future daughter kinomere of each metaphase chromatid with those of adjacent non-homologous chromatids to either side. Thus, four fascicles (SF, 1-4) attach to each metaphase kinomere (K). It is thought that fascicles extend from the kinomere poleward, fray out to let chromosomal fibrils act as traction fibrils against polar fibrils, then regroup to join the adjacent kinomere.

The ionic strength dependence of chromatin folding

1978 ◽  
Vol 36 (2) ◽  
pp. 220-221
Author(s):  
F. Thoma ◽  
TH. Koller
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Ionic Strength ◽  
Specimen Preparation ◽  
Preparation Technique ◽  
Carbon Supports ◽  
Ionic Strength Dependence ◽  
Chromatin Folding ◽  
Strength Dependence ◽  
Preparation Techniques

Under a variety of electron microscope specimen preparation techniques different forms of chromatin appearance can be distinguished: beads-on-a-string, a 100 Å nucleofilament, a 250 Å fiber and a compact 300 to 500 Å fiber.Using a standardized specimen preparation technique we wanted to find out whether there is any relation between these different forms of chromatin or not. We show that with increasing ionic strength a chromatin fiber consisting of a row of nucleo- somes progressively folds up into a solenoid-like structure with a diameter of about 300 Å.For the preparation of chromatin for electron microscopy the avoidance of stretching artifacts during adsorption to the carbon supports is of utmost importance. The samples are fixed with 0.1% glutaraldehyde at 4°C for at least 12 hrs. The material was usually examined between 24 and 48 hrs after the onset of fixation.

Electron Microscopy of Fibroblasts from Myotonic Muscular Dystrophy Patients

1981 ◽  
Vol 39 ◽  
pp. 498-499
Author(s):  
S. E. Miller ◽  
G. B. Hartwig ◽  
R. A. Nielsen ◽  
A. P. Frost ◽  
A. D. Roses
Keyword(s):  
Electron Microscopy ◽  
Muscular Dystrophy ◽  
Genetic Diseases ◽  
Skin Fibroblasts ◽  
Inborn Errors ◽  
Cytoplasmic Inclusions ◽  
Cultured Skin ◽  
Myotonic Muscular Dystrophy ◽  
Glycosaminoglycan Metabolism

Many genetic diseases can be demonstrated in skin cells cultured in vitro from patients with inborn errors of metabolism. Since myotonic muscular dystrophy (MMD) affects many organs other than muscle, it seems likely that this defect also might be expressed in fibroblasts. Detection of an alteration in cultured skin fibroblasts from patients would provide a valuable tool in the study of the disease as it would present a readily accessible and controllable system for examination. Furthermore, fibroblast expression would allow diagnosis of fetal and presumptomatic cases. An unusual staining pattern of MMD cultured skin fibroblasts as seen by light microscopy, namely, an increase in alcianophilia and metachromasia, has been reported; both these techniques suggest an altered glycosaminoglycan metabolism An altered growth pattern has also been described. One reference on cultured skin fibroblasts from a different dystrophy (Duchenne Muscular Dystrophy) reports increased cytoplasmic inclusions seen by electron microscopy. Also, ultrastructural alterations have been reported in muscle and thalamus biopsies from MMD patients, but no electron microscopical data is available on MMD cultured skin fibroblasts.

SEM Evaluation of Fluoride Pretreatment Effects on Acid-Etching of Caries-Like Enamel Lesions In Vitro

1981 ◽  
Vol 39 ◽  
pp. 474-475
Author(s):  
M. John Hicks
Keyword(s):  
Dental Enamel ◽  
Preventive Measure ◽  
Acid Etching ◽  
Distilled Water ◽  
Adhesive Resin ◽  
Fluoride Treatment ◽  
Treatment Groups ◽  
Pretreatment Effects ◽  
Preparation Techniques

Acid-etching of enamel surfaces has been performed routinely to bond adhesive resin materials to sound dental enamel as a caries-preventive measure. The effect of fluoride pretreatment on acid-etching of enamel has been reported to produce inconsistent and unsatisfactory etching patterns. The failure to obtain an adequate etch has been postulated to be due to fluoride precipitation products deposited on the enamel surface. The purpose of this study was to evaluate the effects of fluoride pretreatment on acid-etching of carieslike lesions of human dental enamel.Caries-like lesions of enamel were created in vitro on human molar and premolar teeth. The teeth were divided into two fluoride treatment groups. The specimens were exposed for 4 minutes to either a 2% Sodium Fluoride (NaF) solution or a 10% Stannous Fluoride (SnF2) solution. The specimens were then washed in deionized-distilled water. Each tooth was sectioned into four test regions. This was carried out to compare the effects of various time exposures (0 to 2 minutes) and differing concentrations (10 to 60% w/w) of phosphoric acid (H3PO4) on etching of caries-like lesions. Standard preparation techniques for SEM were performed on the specimens.

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