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Related Experiment Videos

Mechanism for antigen detection on deplasticized epoxy sections

S H Brorson1, F Skjørten

  • 1Department of Pathology, Ullevål Hospital, Oslo, Norway.

Micron (Oxford, England : 1993)
|January 1, 1995
PubMed
Summary

Deplasticizing epoxy sections enhances immunogold labeling, particularly at the periphery of cellular structures. This surface phenomenon improves antigen detection in routinely embedded tissues for pathological laboratories.

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Area of Science:

  • Immunohistochemistry
  • Electron Microscopy
  • Pathology

Background:

  • Epoxy resin embedding is common for ultrastructural preservation.
  • Immunogold labeling is crucial for antigen localization.
  • The effect of deplasticizing epoxy sections on labeling efficiency is not fully understood.

Purpose of the Study:

  • To elucidate the mechanism behind increased immunogold labeling in deplasticized epoxy sections.
  • To compare labeling efficiency between deplasticized and non-deplasticized sections using various gold particle sizes and section thicknesses.
  • To assess antibody penetration depth and its relation to labeling.

Main Methods:

  • Comparative analysis of immunogold labeling ratios on deplasticized versus non-deplasticized epoxy sections.
  • Utilized gold particles of different sizes and varied section thicknesses.
  • Employed the tilt method for analysis.
  • Investigated human kidney, fibrin, and pituitary tissues labeled with specific antibodies.

Main Results:

  • Deplasticizing epoxy sections significantly increased immunogold labeling for amyloid and fibrin, but not for ACTH.
  • Increased labeling was primarily observed at the periphery of structures, with antibody penetration around 30 nm.
  • Labeling ratios were independent of gold particle size and section thickness.
  • No significant difference in labeling was found between deplasticized epoxy and LR-White sections in interior areas.

Conclusions:

  • The enhanced immunogold labeling in deplasticized epoxy sections appears to be a surface phenomenon.
  • Deplasticizing epoxy sections is a valuable method for localizing antigens at the periphery of structures.
  • This technique offers a practical approach for antigen detection in routinely embedded tissues within pathological laboratories.

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