Cytochemical localization of phosphodiesterase by the azo dye simultaneous coupling method

1963 ◽  
Vol 11 (1) ◽  
pp. 70-74 ◽  
Author(s):  
Halina Sierakowska ◽  
D. Shugar
Keyword(s):  
Azo Dye ◽  
Coupling Method ◽  
Cytochemical Localization

scholarly journals CYTOPHOTOMETRIC DETERMINATION OF ALKALINE PHOSPHATASE ACTIVITY OF INDIVIDUAL NEUTROPHILIC LEUKOCYTES WITH A BIOCHEMICALLY CALIBRATED MODEL SYSTEM

1968 ◽  
Vol 16 (11) ◽  
pp. 693-706 ◽  
Author(s):  
M. VAN DER PLOEG ◽  
P. VAN DUIJN
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Azo Dye ◽  
Enzyme Concentration ◽  
Model System ◽  
Coupling Method ◽  
Dye Coupling ◽  
Neutrophilic Leukocytes

A model system consisting of polyacrylamide films into which cell sonicate is incorporated was applied to investigate quantitative aspects of the cytochemical azo dye coupling method for alkaline phosphatase activity in neutrophilic leukocytes. The films were assayed in media containing naphthol AS-MX phosphate and 4-aminodiphenylamine diazonium sulfate. Optimal reaction conditions under which proportionality existed between the amount of reaction product and incubation time or enzyme concentration were determined. Since the enzyme activity in the films could also be measured chemically, using disodium phenyl phosphate as a substrate, a direct relation between the cytochemical and biochemical activity could be established. The azo dye coupling method was applied for the quantification of the enzyme activity. Air-dried microscopic preparations of exudate neutrophils were stained and the amount of dye formed in the cytoplasm of individual leukocytes was measured with a cytospectrophotometer based on the two-wavelength principle. By reference to the relation between biochemical and cytochemical activities in the model films, the cytochemically determined enzyme activity could be expressed in biochemical units. Independently, the average alkaline phosphatase activity per cell was determined by direct biochemical assay of the leukocyte suspension. The results showed that about 60% of the biochemically assayed activity in sonicate was demonstrated in the cell preparations by the cytochemical staining method. The discrepancy between the results of the two methods is discussed. The applicability of the model system for elucidation of the relationship between the results of cytochemical and biochemical enzyme determination in cells is stressed.

scholarly journals A cytochemical stain for glutathione in rat hepatocytes cultured on plastic.

1987 ◽  
Vol 35 (2) ◽  
pp. 271-274 ◽  
Author(s):  
A Larrauri ◽  
P López ◽  
M J Gómez-Lechón ◽  
J V Castell
Keyword(s):  
Azo Dye ◽  
Room Temperature ◽  
Cultured Cells ◽  
Rat Hepatocytes ◽  
Thiol Groups ◽  
Cytochemical Localization ◽  
Sh Groups ◽  
Acetone Water ◽  
Mercury Orange ◽  
Orange Fluorescence

Thiol groups of glutathione react with the organomercurial azo dye mercury orange at a faster rate than with -SH groups of proteins. This property makes possible visualization of glutathione in cells without appreciable interference from other -SH groups. To render this method useful for cytochemical localization of glutathione in plastic cultured cells, it was necessary to adapt this reaction to the specific characteristics of the biological samples to be assayed. First, the choice of a solvent that would allow a convenient solubility of the dye and at the same time be compatible with the plastic culture plate was crucial. Second, to avoid diffusion of glutathione out of the cell the procedure for staining cells was also important. Satisfactory results were obtained after 30-40 sec reaction with 50 microM mercury orange in acetone/water 9:1, v/v, at room temperature. Glutathione-mercury orange complexes exhibited orange fluorescence on excitation with blue light. No diffusion of glutathione out of the cells was observed, and the hepatocytes stained with the dye showed orange fluorescence which paralleled their glutathione content.

Initial Polymerization Sites Indicated During Mitochondrial Oxidation of Diaminobenzidine after Toluene Treatment

1977 ◽  
Vol 35 ◽  
pp. 408-409
Author(s):  
W. A. Shannon ◽  
M. A. Matlib
Keyword(s):  
Membrane Permeability ◽  
Cytochrome Oxidase ◽  
Rat Liver ◽  
Precise Definition ◽  
Cytochemical Localization ◽  
Oxidative Reaction ◽  
Mitochondrial Oxidation ◽  
Definition Of ◽  
Exogenous Substrates

Numerous studies have dealt with the cytochemical localization of cytochrome oxidase via cytochrome c. More recent studies have dealt with indicating initial foci of this reaction by altering incubation pH (1) or postosmication procedure (2,3). The following study is an attempt to locate such foci by altering membrane permeability. It is thought that such alterations within the limits of maintaining morphological integrity of the membranes will ease the entry of exogenous substrates resulting in a much quicker oxidation and subsequently a more precise definition of the oxidative reaction.The diaminobenzidine (DAB) method of Seligman et al. (4) was used. Minced pieces of rat liver were incubated for 1 hr following toluene treatment (5,6). Experimental variations consisted of incubating fixed or unfixed tissues treated with toluene and unfixed tissues treated with toluene and subsequently fixed.

Cytochemical Localization of Polysaccharides in Diplodia Maydis With Thiosemicarbazide : Evaluation of Three Fixatives For Pa:Tsc:Os Reaction

1976 ◽  
Vol 34 ◽  
pp. 48-49 ◽  
Author(s):  
Judith A. Murphy ◽  
Mary R. Thompson ◽  
A.J. Pappelis
Keyword(s):  
Reaction Center ◽  
Osmium Tetroxide ◽  
Periodic Acid ◽  
Cytochemical Localization ◽  
Thin Sections ◽  
Selective Reaction ◽  
Amorphous Character

In an attempt to identify polysaccharide components in thin sections of D. maydis, procedures were employed such that a PAS localization could be carried out. Three different fixatives were evaluated ie. glutaraldehyde, formaldehyde and paraformaldehyde. These were used in conjunction with periodic acid (PA), thiosemicarbazide(TSC), and osmium tetroxide(Os) to localize polysaccharides in V. maydis using a pre-embedded reaction procedure. Polysaccharide localization is based on the oxidation of vic-glycol groups by PA, and the binding of TSC as a selective reaction center for the formation of osmium black. The reaction product is sufficiently electron opaque, insoluble in lipids, not altered when tissue is embedded, and has a fine amorphous character.

Effect of Nitrogen Sources on the AZO Dye Decolourization

2012 ◽  
Vol 2 (7) ◽  
pp. 424-426
Author(s):  
Suchita Dahiwade ◽  
◽  
Dr. A. O. Ingle Dr. A. O. Ingle ◽  
Dr. S. R. Wate Dr. S. R. Wate
Keyword(s):  
Azo Dye ◽  
Nitrogen Sources ◽  
Dye Decolourization

Determination of the food azo dye carmoisine vs chlorpheniramine maleate ion associate structure

2014 ◽  
Vol 0 (3(79)) ◽  
pp. 34-37
Author(s):  
A. S. Materiienko ◽  
V. O. Grudko ◽  
V. A. Khanin ◽  
V. A. Georgiyants
Keyword(s):  
Azo Dye ◽  
Chlorpheniramine Maleate ◽  
Ion Associate

Use of a sequencing batch biofilter for degradation of azo dyes (acids and bases)

2000 ◽  
Vol 42 (5-6) ◽  
pp. 329-336 ◽  
Author(s):  
M. Quezada ◽  
I. Linares ◽  
G. Buitrón
Keyword(s):  
Azo Dyes ◽  
Azo Dye ◽  
Degradation Rate ◽  
Removal Rate ◽  
Textile Effluent ◽  
Colour Removal ◽  
Substrate Degradation ◽  
Acids And Bases ◽  
Removal Efficiencies ◽  
Basic Red 46

The degradation of azo dyes in an aerobic biofilter operated in an SBR system was studied. The azo dyes studied were Acid Red 151 and a textile effluent containing basic dyes (Basic Blue 41, Basic Red 46 and 16 and Basic Yellow 28 and 19). In the case of Acid Red 151 a maximal substrate degradation rate of 288 mg AR 151/lliquid·d was obtained and degradation efficiencies were between 60 and 99%. Mineralization studies showed that 73% (as carbon) of the initial azo dye was transformed to CO2 by the consortia. The textile effluent was efficiently biodegraded by the reactor. A maximal removal rate of 2.3 kg COD/lliquid·d was obtained with removal efficiencies (as COD) varying from 76 to 97%. In all the cycles the system presented 80% of colour removal.

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