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Staining tissue with methacrylate casts for light microscopy

M Sekosan1, D E Schraufnagel

  • 1Department of Pathology, University of Illinois at Chicago 60612-7323, USA.

Scanning
|December 19, 1998
PubMed
Summary
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Histologic stains improve light microscopy of methacrylate casts in human lung microcirculation. The Ziehl-Neelsen stain best accentuates the cast, enhancing visualization of capillaries and cells in both normal and diseased lung tissue.

Area of Science:

  • Histology
  • Microscopy
  • Pulmonary Medicine

Background:

  • Scanning electron microscopy (SEM) and light microscopy (LM) are established methods for studying microcirculation.
  • Methacrylate casts are used in SEM but are transparent and difficult to appreciate with LM.
  • Improved LM visualization of methacrylate casts could enhance correlation between SEM and LM findings.

Purpose of the Study:

  • To evaluate different histologic stains for their ability to accentuate methacrylate casts in human lung tissue.
  • To determine if staining methacrylate casts improves the appreciation of microvascular structures and cellular detail under LM.
  • To assess the utility of stained casts for correlating SEM and LM findings.

Main Methods:

  • Human lung tissue (surgically removed and autopsied) was cast with methyl methacrylate.

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  • Cast lung tissue was processed using routine LM methods.
  • Various stains were applied, including hematoxylin and eosin, Masson trichrome, elastic-van Gieson, Grocott methenamine silver, Brown-Brennan, and Ziehl-Neelsen.

    • Main Results:

    • The Ziehl-Neelsen stain effectively stained the methacrylate cast red, with optimal results achieved without heating.
    • Acid-fast staining of cast lung tissue provided superior visualization of capillaries and cells in both normal and diseased lungs compared to hematoxylin and eosin.
    • Methacrylate casts in lung tissue can be successfully processed for routine LM.

    Conclusions:

    • The Ziehl-Neelsen technique is highly effective for staining methacrylate casts in lung tissue for LM.
    • This method enhances the visualization of microcirculation and cellular details, improving the correlation between SEM and LM.
    • Stained methacrylate lung casts offer a valuable tool for assessing microvascular filling and correlating findings from different microscopic techniques.