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Alternative staining methods for Lowicryl sections.

R A Horowitz1, C L Woodcock

  • 1Department of Zoology, University of Massachusetts, Amherst.

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
|January 1, 1992
PubMed
Summary
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New staining methods using hydrophilic resin Lowicryl K11M offer improved contrast and specificity for biological samples. These techniques enhance visualization of cellular structures compared to traditional aqueous uranyl and lead salts staining.

Area of Science:

  • Electron microscopy
  • Cell biology
  • Biochemistry

Background:

  • Traditional aqueous uranyl and lead salts (UA-Pb) staining in electron microscopy exhibits limitations in granularity and specificity.
  • The hydrophilic resin Lowicryl K11M offers potential for improved contrast and preservation of cellular ultrastructure.

Purpose of the Study:

  • To identify and evaluate novel staining techniques for electron microscopy using Lowicryl K11M.
  • To compare the efficacy of new stains and combinations against conventional UA-Pb staining.

Main Methods:

  • Evaluation of various stain combinations with Lowicryl K11M resin.
  • Testing on model biological specimens: tobacco mosaic virus (TMV), starfish sperm, and cultured mouse fibroblasts.
  • Assessment of staining characteristics including granularity, specificity, and contrast range.

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Main Results:

  • Specific staining of nucleic acids was achieved using osmium ammine-B at pH 1.5.
  • Combinations of uranyl acetate with barium, manganese, tungsten, molybdenum, and vanadium salts provided enhanced contrast for protein-containing components.
  • Novel fibrillar and particulate structures within the nucleus were visualized that were not apparent with UA-Pb staining.

Conclusions:

  • New staining protocols utilizing Lowicryl K11M resin and specific metal salts significantly improve electron microscopic visualization of cellular components.
  • These advanced staining methods offer superior specificity and contrast for nucleic acids and proteins, revealing previously unobserved ultrastructural details.