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Abstract

In the past four years there has been a distinct tendency to abandon the earlier methods of enzyme histochemistry that were characterized by a general and diffuse cytoplasmic localization and to direct more interest and attention to methods capable of demonstrating mitochondria or other organelles. Disenchantment with former methods has undoubtedly been accelerated by the perfection of methods for the dehydrogenases, which demonstrate mitochondria clearly and readily [38, 59, 60]. It has been finally recognized, that much of the diffuseness obtained with hydrolytic enzymes was due to the soluble and diffusible component of the enzymes (lyoenzyme). By using various methods of fixation this component could be effectively allowed to diffuse from the tissue leaving behind for histochemical demonstration the more precisely localized, less soluble component of the enzymes (desmoenzyme) [36], made even more insoluble by appropriate fixation. The use of better fixation, better substrates and enzyme inhibitors has been instrumental in delineating the droplet form of some hydrolytic enzymes, and these localizations have been correlated with de Duvr’s interesting and important biochemical work on the lysosomes [11, 23, 40]. Most popular have become methods for various dehydrogenases using the ditetrazolium hydrogen acceptor, nitro-BT [57] and for cytochrome oxidase using analogues of phenylene diamine and naphtholic derivatives [7, 33]. The methods for the hydrolytic enzymes have been improved by using formalinfixed tissue [51] and substrates and coupling agents that yield insoluble and substantive dye pigments, such for example as the demonstration of esterase with fast blue BB and naphthol-AS acetate, a substrate first proposed by Dr. Gomori [16]. Much effort has been directed with success toward proper fixation in order to preserve organelle morphology [12, 18, 59, 60] while maintaining significant enzymatic activity and at the same time allowing diffusible enzyme to leave the tissue block.

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Copyright information

© Springer-Verlag Berlin Heidelberg 1964

Authors and Affiliations

  • A. M. Seligman
    • 1
    • 2
  • A. G. E. Pearse
    • 3
    • 4
  • William A. Jensen
    • 5
  • W. Maurer
    • 6
  • Hewson Swift
    • 7
  • R. Vendrely
    • 8
  • Oliver H. Lowry
    • 9
    • 10
  • Erik Zeuthen
    • 11
  1. 1.Departments of Surgery, Sinai Hospital of Baltimore and The Johns HopkinsUniversity School of MedicineBaltimoreUSA
  2. 2.Sinai Hospital of BaltimoreBaltimore 15USA
  3. 3.Postgraduate Medical School of LondonLondon, W 12England
  4. 4.University of LondonEngland
  5. 5.Department of BotanyUniversity of CaliforniaBerkeleyUSA
  6. 6.Institut für med. IsotopenforschungUniversität KölnGermany
  7. 7.Dept. of ZoologyThe University of ChicagoChicagoUSA
  8. 8.Institut de Recherches Scientifiques sur le CancerVillejuif (Seine)France
  9. 9.Dept. of PharmacologyWashington Univ. School of MedicineSt. LouisUSA
  10. 10.Dept. of Pharmacology and the Beaumont-May Institute of NeurologyWashington University School of MedicineSt. LouisUSA
  11. 11.The Biological Institute of the Carlsberg FoundationCopenhagen NDenmark

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