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Cytochemical Techniques for the Subcellular Localization of Enzymes in Microorganisms

  • Martha J. Powell

Abstract

Methods to localize intracellular sites of enzymes with electron microscopy grew directly from light microscopic histochemistry. Techniques, such as Gomori’s (1952) for acid phosphatase activity, which had as end products heavy metal ions with sufficient mass to scatter electrons, were directly applicable for electron microscopy if noncoagulative fixatives were used. Introduction of catalytic osmiophilic polymer generation (Hanker et al., 1972a) made substrates previously valuable only for light microscopy useful for electron microscopy. In the most widely used strategies for electron microscopic localization of enzymes in microorganisms, enzymes are not viewed directly, but reactions with end products impart electron density to the sites of enzyme activity. There are numerous strategies for the cytochemical localization of enzymes, and this chapter will emphasize three of these: (1) ion capture and precipitation of products; (2) ferricyanide reduction and product amplification; and (3) oxidative polymerization of diaminobenzidine. Thus, this discussion is not exhaustive of all techniques used for microorganisms but is illustrative of rationales used in attempting to identify sites of enzyme activities.

Keywords

Acid Phosphatase Reaction Medium Acid Phosphatase Activity Aryl Sulfatase Ultrastructural Localization 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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© Plenum Press, New York 1986

Authors and Affiliations

  • Martha J. Powell
    • 1
  1. 1.Department of BotanyMiami UniversityOxfordUSA

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