X-Ray Analysis of Cytochemical Reaction Products in Synaptic Areas

1976 ◽  
Vol 34 ◽  
pp. 6-7 ◽  
Author(s):  
J. Wood
Keyword(s):  
Electron Microscopic Level ◽  
Junctional Complex ◽  
Reaction Products ◽  
Electron Microscopic ◽  
Cytochemical Localization ◽  
X Ray ◽  
The Past ◽  
Electron Dense Deposit ◽  
Dense Deposit ◽  
Cytochemical Reaction

Specific cytochemical reactions have been instrumental in the illucidation of compounds within tissues, whether these compounds are hormones, enzymes, or molecules, such as certain nerve transmitter agents. Many cytochemical reaction products depend upon some complex, which is an electron dense deposit. Several types of cytochemical procedures can be used to visualize agents related to synaptic transmission at the junctional complex. One method which has been used with considerable success has been the cytochemical localization of biogenic amines (BAs), i.e., norepinephrine (NE) and dopamine (DA). For the past few years, a chrome complex formed with certain BAs and glutaraldehyde has been utilized to localize BAs at the electron microscopic level and the specificity of the reaction has been verified biochemically.

X-Ray Microanalysis of Nickel and Cobalt Intensified Peroxidase- Antiperoxidase For Verification of Reaction Sites

1985 ◽  
Vol 43 ◽  
pp. 468-469
Author(s):  
A. Angel ◽  
K. Miller ◽  
V. Seybold ◽  
R. Kriebel
Keyword(s):  
Reaction Mechanisms ◽  
Visual Discrimination ◽  
Osmium Tetroxide ◽  
Ultrastructural Level ◽  
Microscopic Level ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
X Ray ◽  
Insoluble Polymer ◽  
Cytochemical Reaction

Localization of specific substances at the ultrastructural level is dependent on the introduction of chemicals which will complex and impart an electron density at specific reaction sites. Peroxidase-antiperoxidase(PAP) methods have been successfully applied at the electron microscopic level. The PAP complex is localized by addition of its substrate, hydrogen peroxide and an electron donor, usually diaminobenzidine(DAB). On oxidation, DAB forms an insoluble polymer which is able to chelate with osmium tetroxide becoming electron dense. Since verification of reactivity is visual, discrimination of reaction product from osmiophillic structures may be difficult. Recently, x-ray microanalysis has been applied to examine cytochemical reaction precipitates, their distribution in tissues, and to study cytochemical reaction mechanisms. For example, immunoreactive sites labelled with gold have been ascertained by means of x-ray microanalysis.

scholarly journals Analytical electron microscopic evaluation of the effects of paraformaldehyde pretreatment on the reaction of glutaraldehyde with biogenic amines.

1982 ◽  
Vol 30 (5) ◽  
pp. 481-486 ◽  
Author(s):  
R E McClung ◽  
J Wood
Keyword(s):  
Electron Microscopy ◽  
Analytical Electron Microscopy ◽  
Microscopic Level ◽  
Electron Microscopic Level ◽  
Spectrographic Analysis ◽  
Electron Microscopic ◽  
X Ray ◽  
Microscopic Evaluation ◽  
Analytical Electron ◽  
Dense Product

Analytical electron microscopy was used to determine the quantitative effects of paraformaldehyde pretreatment on the formation of the biogenic amine-glutaraldehyde-chrome complex. Pretreatment with paraformaldehyde prevented the glutaraldehyde-chrome reaction with norepinephrine in the rat adrenal medulla. In contrast to the effect of paraformaldehyde on norepinephrine, pretreatment did not prevent the chrome reaction in serotonin-containing argentaffin cells of the gut. X-Ray energy spectrographic analysis revealed a significant decrease in chrome content in the paraformaldehyde treated tissue, but sufficient chrome did react to produce an electron-dense product. Thus by treating tissue with paraformaldehyde prior to the glutaraldehyde chrome procedure, serotonergic sites may be differentiated from catecholaminergic areas at the electron microscopic level.

scholarly journals Biochemical Properties and Cytochemical Localization of Ouabain-insensitive, Potassium-dependent p-Nitrophenylphosphatase Activity in Rat Atrial Myocytes

1997 ◽  
Vol 45 (2) ◽  
pp. 177-187 ◽  
Author(s):  
Vadim S. Zinchuk ◽  
Toshihiro Kobayashi ◽  
Eva Garcia del Saz ◽  
Harumichi Seguchi
Keyword(s):  
Ventricular Myocytes ◽  
Reaction Medium ◽  
Biochemical Properties ◽  
Atrial Myocytes ◽  
Electron Microscopic ◽  
Cytochemical Localization ◽  
X Ray ◽  
T Tubules ◽  
P Type ◽  
Triton X

Enzyme activity that represents ouabain-insensitive, potassium-dependent p-nitrophenylphosphatase (p-NPPase) was assessed in rat atrial myocytes by biochemical and cytochemical procedures. No activity was detected in parallel experiments with ventricular myocytes. Fixed tissues were incubated in a reaction medium containing Tricine buffer, p-nitrophenylphosphate (p-NPP), KCl, MgCl2, CaCl2, CeCl3, Triton X-100, levamisole, and ouabain. Final pH was adjusted to 7.5. Biochemical studies showed that accumulation of p-nitrophenol in the medium was increased proportionally in accordance with the amount of incubated tissue. This activity was optimal with incubation at pH 7.5 and in the presence of KCl. Approximately 70% of the enzyme was inhibited by 2 mM CeCl3. Electron microscopic observations revealed reaction product (RP) at sites of ouabain-insensitive, potassium-dependent p-NPPase activity as electron-dense precipitate localized at the inner surface of the plasma membrane and at the T-tubules of atrial myocytes. Control experiments indicated that the activity was strongly inhibited by sodium orthovanadate and was repressed by omeprazole and 1,3-dicyclohexylcarbodiimide. X-ray microanalysis confirmed the presence of cerium within the cytochemical RP. The ouabain-insensitive, K-dependent p-NPPase activity detected in the present study is considered to be an isoform of a P-type, H-transporting, K-dependent adenosine triphosphatase (H,K-ATPase).

scholarly journals D-aspartate oxidation by rat and bovine renal peroxisomes: an electron microscopic cytochemical study.

1990 ◽  
Vol 38 (9) ◽  
pp. 1377-1381 ◽  
Author(s):  
M E Beard
Keyword(s):  
Amino Acid ◽  
Oxidase Activity ◽  
Electron Microscopic Level ◽  
Acid Oxidase ◽  
Electron Microscopic ◽  
Amino Acid Oxidase ◽  
Cytochemical Localization ◽  
Peroxisomal Enzyme ◽  
Cytochemical Study ◽  
Biochemical Studies

D-amino acid oxidase, a peroxisomal enzyme, and D-aspartate oxidase, a potential peroxisomal enzyme, share biochemical attributes. Both produce hydrogen peroxide in flavin-requiring oxidative reactions. Such similarities suggest that D-aspartate oxidase may also be localized to peroxisomes. Definitive identification of D-aspartate oxidase as a peroxisomal enzyme depends, however, on visualization at the electron microscopic level. Using incubation conditions shown to be specific for the enzyme in biochemical studies, this report extends the cytochemical localization of D-amino acid oxidase to bovine renal peroxisomes, and shows that D-aspartate can be oxidized by rat and bovine renal peroxisomes. An unexpected finding was the sensitivity of both D-amino acid oxidase activity (proline specific) and D-aspartate oxidase activity to inhibition by agents used in biochemical studies to discriminate between the two enzyme activities. Therefore, it is possible that, in the cytochemical system used in this study, (a) either D-proline and D-aspartate are substrates for only one enzyme or (b) the two enzymes have additional overlapping biochemical properties.

scholarly journals Combination of horseradish peroxidase and lucifer yellow staining for selective labeling of neurons at the electron microscopic level.

1991 ◽  
Vol 39 (11) ◽  
pp. 1579-1583 ◽  
Author(s):  
L Nonnotte ◽  
A Buisson ◽  
F Nagy ◽  
M Moulins
Keyword(s):  
Horseradish Peroxidase ◽  
Lucifer Yellow ◽  
Electron Microscopic Level ◽  
Reaction Products ◽  
Stomatogastric Nervous System ◽  
Electron Microscopic ◽  
Double Labeling ◽  
Selective Labeling ◽  
Experimental Neuroanatomy ◽  
Labeling Method

We have developed a new double labeling method for electron microscopy to characterize selectively two physiologically identified neurons on the same preparation. The stomatogastric nervous system of crustaceans was used to test the distinguishing staining characteristics of the two labels. Neurons were labeled on one side with horseradish peroxidase (HRP) and on the other side with Lucifer yellow (LY). After blue light irradiation of the tissue in the presence of diaminobendizine, the two labeled neurons could be easily observed and discriminated on the same section by the two different reaction products. This simple technique of double labeling is useful in experimental neuroanatomy for the detailed study of synaptic relationships.

scholarly journals IDENTIFICATION OF CYTOCHEMICAL REACTION PRODUCTS BY SCANNING X-RAY EMISSION MICROANALYSIS

1962 ◽  
Vol 15 (3) ◽  
pp. 427-435 ◽  
Author(s):  
Arthur J. Hale
Keyword(s):  
Alkaline Phosphatase ◽  
Staining Method ◽  
Reaction Products ◽  
Elementary Composition ◽  
Cytochemical Staining ◽  
Emission Analysis ◽  
X Ray ◽  
Yield Information ◽  
Cytochemical Reaction

The primary, secondary, and final reaction products of the cytochemical staining method for alkaline phosphatase have been demonstrated by x-ray emission microanalysis. The advantages and limitations of the technique in measuring reaction products are discussed. Scanning x-ray emission analysis provides a rapid way of detecting and measuring the amounts of individual elements in cytochemical reaction products. It has a resolution of 1 µ and a sensitivity of 0.01 per cent (w/v). It will provide information on the elementary composition of the products in situ and will permit study of the efficiency of conversion of one product to another. It will also yield information on the diffusion of reaction products in the tissue.

scholarly journals Relationship between IgA Nephropathy and Porphyromonas gingivalis; Red Complex of Periodontopathic Bacterial Species

2021 ◽  
Vol 22 (23) ◽  
pp. 13022
Author(s):  
Yasuyuki Nagasawa ◽  
Ryota Nomura ◽  
Taro Misaki ◽  
Seigo Ito ◽  
Shuhei Naka ◽  
...  
Keyword(s):  
Iga Nephropathy ◽  
Porphyromonas Gingivalis ◽  
Bacterial Species ◽  
Electron Microscopic Observation ◽  
Nasal Administration ◽  
Mice Model ◽  
Electron Microscopic ◽  
Electron Dense Deposit ◽  
Periodontal Bacteria ◽  
Dense Deposit

IgA nephropathy (IgAN) has been considered to have a relationship with infection in the tonsil, because IgAN patients often manifest macro hematuria just after tonsillitis. In terms of oral-area infection, the red complex of periodontal bacteria (Porphyromonas gingivalis (P. gingivalis), Treponema denticol (T. denticola) and Tannerella forsythia (T. forsythia)) is important, but the relationship between these bacteria and IgAN remains unknown. In this study, the prevalence of the red complex of periodontal bacteria in tonsil was compared between IgAN and tonsillitis patients. The pathogenicity of IgAN induced by P. gingivalis was confirmed by the mice model treated with this bacterium. The prevalence of P. gingivalis and T. forsythia in IgAN patients was significantly higher than that in tonsillitis patients (p < 0.001 and p < 0.05, respectively). A total of 92% of tonsillitis patients were free from red complex bacteria, while only 48% of IgAN patients had any of these bacteria. Nasal administration of P. gingivalis in mice caused mesangial proliferation (p < 0.05 at days 28a nd 42; p < 0.01 at days 14 and 56) and IgA deposition (p < 0.001 at day 42 and 56 after administration). Scanning-electron-microscopic observation revealed that a high-density Electron-Dense Deposit was widely distributed in the mesangial region in the mice kidneys treated with P. gingivalis. These findings suggest that P. gingivalis is involved in the pathogenesis of IgAN.

The Use of Advanced Analytical Techniques for Characterization of the Chemical, Physical, and Crystallographic Nature of a Surface

1973 ◽  
Vol 31 ◽  
pp. 132-133 ◽  
Author(s):  
R. E. Herfert
Keyword(s):  
Surface Characterization ◽  
Analytical Techniques ◽  
X Ray Diffraction ◽  
Depth Of Penetration ◽  
X Ray ◽  
The Past ◽  
Transmission Electron ◽  
Scanning Electron

Studies of the nature of a surface, either metallic or nonmetallic, in the past, have been limited to the instrumentation available for these measurements. In the past, optical microscopy, replica transmission electron microscopy, electron or X-ray diffraction and optical or X-ray spectroscopy have provided the means of surface characterization. Actually, some of these techniques are not purely surface; the depth of penetration may be a few thousands of an inch. Within the last five years, instrumentation has been made available which now makes it practical for use to study the outer few 100A of layers and characterize it completely from a chemical, physical, and crystallographic standpoint. The scanning electron microscope (SEM) provides a means of viewing the surface of a material in situ to magnifications as high as 250,000X.

Silver Staining of Pancreatic Islets as Revealed by Electron Microscopy

1967 ◽  
Vol 25 ◽  
pp. 44-45
Author(s):  
Kazuaki Misugi ◽  
Nobuko Misugi ◽  
Hiroshi Yamada
Keyword(s):  
Pancreatic Islet ◽  
Silver Staining ◽  
Islet Cells ◽  
Severe Hypoglycemia ◽  
Granular Cell ◽  
Electron Microscopic Level ◽  
Staining Reaction ◽  
Electron Microscopic ◽  
Pancreatic Islet Cell ◽  
D Cell

The authors had described the fine structure of a type of pancreatic islet cell, which appeared different from typical alpha and beta cells, and tentatively considered that this third type of granular cell probably represents the D cell (Figure 1).Since silver staining has been widely used to differentiate different types of pancreatic islet cells by light microscopy, an attempt to examine this staining reaction at the electron microscopic level was made.Material and Method: Surgically removed specimens from three infants who suffered from severe hypoglycemia were used. The specimens were fixed and preserved in 20% neutral formalin. Frozen sections, 30 to 40 micron thick, were prepared and they were stained by Bielschowsky's method as modified by Suzuki (2). The stained sections were examined under a microscope and islet tissues were isolated. They were fixed in 1% osmium tetroxide in phosphate buffer for one hour and embedded in Epon 812 following dehydration through a series of alcohols and propylene oxide.

A simplified method of emulsion coating and development for quantitative electron microscopic radioautography

1978 ◽  
Vol 36 (2) ◽  
pp. 64-65
Author(s):  
K. Yoshida ◽  
F. Murata ◽  
S. Ohno ◽  
T. Nagata
Keyword(s):  
Mass Production ◽  
Identical Condition ◽  
Microscopic Level ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
Simplified Method ◽  
Complicated Process ◽  
Critical Problems ◽  
Electron Microscopic Radioautography ◽  
Quantitative Radioautography

IntroductionSeveral methods of mounting emulsion for radioautography at the electron microscopic level have been reported. From the viewpoint of quantitative radioautography, however, there are many critical problems in the procedure to produce radioautographs. For example, it is necessary to apply and develop emulsions in several experimental groups under an identical condition. Moreover, it is necessary to treat a lot of grids at the same time in the dark room for statistical analysis. Since the complicated process and technical difficulties in these procedures are inadequate to conduct a quantitative analysis of many radioautographs at once, many factors may bring about unexpected results. In order to improve these complicated procedures, a simplified dropping method for mass production of radioautographs under an identical condition was previously reported. However, this procedure was not completely satisfactory from the viewpoint of emulsion homogeneity. This paper reports another improved procedure employing wire loops.

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