Photographic microdensitometry for evaluation of acid phosphatase activity at the electron microscope level

1986 ◽  
Vol 18 (9) ◽  
pp. 481-486
Author(s):  
R. L. Cabrini ◽  
M. E. Itoiz ◽  
R. E. Alvarez ◽  
S. C. Orrea
1965 ◽  
Vol 25 (1) ◽  
pp. 23-41 ◽  
Author(s):  
Lois Withrow Tice ◽  
Russell J. Barrnett

This paper reports the synthesis of 14 diazophthalocyanins containing Mg, Cu, or Pb as the chelated metal. To assess the usefulness of these compounds for fine structural cytochemistry, the relative coupling rates with naphthols were tested as well as the solubility of the resulting azo dyes. Three of the diazotates were reacted with tissue proteins in aldehyde-fixed material, and the density increases thus produced were compared in the electron microscope with those produced by staining similarly fixed material with the phthalocyanin dye, Alcian Blue. Finally, one of the diazotates was used as a capture reagent for the demonstration of the sites of acid phosphatase activity with the electron microscope.


Parasitology ◽  
1968 ◽  
Vol 58 (2) ◽  
pp. 371-375 ◽  
Author(s):  
David A. Erasmus

A combination of histochemical and electron microscope techniques have demonstrated, in Cyathocotyle bushiensis, alkaline phosphatase activity in the matrix of the tegument, in the distal and basal plasma membranes of the tegument, in the wall of the ducts extending from the adhesive organ gland cells and in the wall of the adhesive organ microvilli. Acid phosphatase activity was much stronger and was present in the tegument matrix and in the granular component of the secretion from the adhesive organ gland cells. Strong acid phosphatase activity was also present in the cisternae of the endoplasmic reticulum of the adhesive organ gland cells.I am greatly indebted to Professor Brough for the excellent facilities available within this department. I also wish to thank Professor J. Sinclair (Department of Mining) for electron microscope facilities extended to me in the early stages of this investigation, and to Mr W. Henderson, Mr T. Davies and Miss M. Williams for their invaluable assistance. The purchase of the Huxley ultramicrotome, coating unit and an AEI EM 6 electron microscope was made possible by a grant from the Science Research Council.


1972 ◽  
Vol 52 (2) ◽  
pp. 465-477 ◽  
Author(s):  
Bertram Sacktor ◽  
Yoshio Shimada

Mitochondria from flight muscle of aging blowflies, Phormia regina, were examined morphologically and biochemically with the electron microscope. An age-dependent degeneration of the mitochondria that is characterized, in part, by the reorganization of the inner membrane into myelin-like whorls has been found. The concentric rings increase in size and number, eventually replacing the normal cristal conformation. Glycogen rosettes are frequently seen in the center of the whorl and may represent the intrusion into the mitochondria of the glycogen in the cytoplasmic matrix of the muscle. The degenerating mitochondria are not associated with lysosomal activity, as indicated by the absence of acid phosphatase. An intense acid phosphatase activity is noted, however, in the dyad, comprising elements of the T system and sarcoplasmic reticulurn. Cytochrome oxidase is active in the ultrastructurally intact portion of the mitochondrion but activity is not evident in that part of the mitochondrion that has undergone morphological change. Thus, the ultrastructural degradation of the mitochondria is correlated with a decrease in biochemical function. This suggests a correspondence between a decrease in the bioenergetic capacity of the flight muscle and a decline in the ability of the aged insect to fly.


1971 ◽  
Vol 8 (3) ◽  
pp. 727-733
Author(s):  
R. B. KNOX ◽  
J. HESLOP-HARRISON

Acid phosphatase has been localized in the wall of the pollen grain of Crocus vernus Wulf at the electron-microscope level by a method using 2-naphthyl thiol phosphate as substrate in a simultaneous coupling reaction with fast blue BBN at pH 5.0, the product being given electron opacity by osmication. Activity was found to be concentrated mainly in the intine, and to be associated with ribbon-like or filamentous inclusions believed to be proteinaceous on the basis of other criteria. Some activity was also detectable in the interstices of the exine. The observations confirm the general interpretation of the distribution of wall-held enzyme based upon light-microscopic cytochemistry, and provide the resolution necessary to establish unambiguously that they are associated with protein layers inserted during intine growth.


Author(s):  
O. T. Minick ◽  
E. Orfei ◽  
F. Volini ◽  
G. Kent

Hemolytic anemias were produced in rats by administering phenylhydrazine or anti-erythrocytic (rooster) serum, the latter having agglutinin and hemolysin titers exceeding 1:1000.Following administration of phenylhydrazine, the erythrocytes undergo oxidative damage and are removed from the circulation by the cells of the reticulo-endothelial system, predominantly by the spleen. With increasing dosage or if animals are splenectomized, the Kupffer cells become an important site of sequestration and are greatly hypertrophied. Whole red cells are the most common type engulfed; they are broken down in digestive vacuoles, as shown by the presence of acid phosphatase activity (Fig. 1). Heinz body material and membranes persist longer than native hemoglobin. With larger doses of phenylhydrazine, erythrocytes undergo intravascular fragmentation, and the particles phagocytized are now mainly red cell fragments of varying sizes (Fig. 2).


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