A simple post-embedding system for the rapid demonstration of tissue antigens under the electron microscope

Histochem J. 1983 Jun;15(6):543-55. doi: 10.1007/BF01954145.

Abstract

A simple and versatile technique for the preparation of ultra-thin sections, which can be stained immunohistochemically directly on electron microscope grids, is presented. An anti-hapten immunoperoxidase procedure has been adapted for use on tissue fixed in a purified monomeric glutaraldehyde--picric acid mixture, and embedded in 'L R White', a recently formulated plastic resin. This plastic tolerates the use of partial dehydration of tissue, resulting in higher antigenic yields. In addition, no etching of ultra-thin sections is necessary, and the whole immunostaining procedure can be completed in less than 2 h. A comparison of commonly used fixatives is discussed. High-resolution micrographs showing general staining (uranyl acetate--lead citrate) of rat pancreas, and immunostaining of insulin and TSH in storage granules in perfusion-fixed rat tissue and of lambda-chain immunoreactive cells in immersion-fixed human tonsil are included as examples.

MeSH terms

  • Animals
  • Humans
  • Immunoglobulin lambda-Chains / metabolism
  • Immunologic Techniques*
  • Insulin / metabolism
  • Islets of Langerhans / ultrastructure
  • Microscopy, Electron / methods*
  • Palatine Tonsil / ultrastructure
  • Pancreas / ultrastructure
  • Plastics
  • Rats
  • Thyrotropin / metabolism

Substances

  • Immunoglobulin lambda-Chains
  • Insulin
  • Plastics
  • Thyrotropin