Vapor fixation for immunocytochemistry and X-ray microanalysis on cryoultramicrotome sections

J Histochem Cytochem. 1984 Jun;32(6):636-42. doi: 10.1177/32.6.6373915.

Abstract

Several organic and inorganic vapor fixatives have been tested for their ability to stabilize the ultrastructure of freeze-dried thin cryosections. The vapors from osmium tetroxide and dry formaldehyde gave a good preservation of the ultrastructure. Fixation in formaldehyde vapor preserved the immunoreactivity of alpha-amylase in exocrine pancreas, as was demonstrated with an indirect labeling technique using anti-alpha-amylase and protein A-gold. A major advantage of the use of vapor fixation is that cryosections from a specimen of fresh-frozen tissue can be used for immunocytochemistry as well as for X-ray microanalysis, as was demonstrated for the exocrine pancreas. This opens the possibility of localizing atomic species (X-ray microanalysis) and molecular species (immunocytochemistry) at the subcellular level on thin cryosections from the same tissue block.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Electron Probe Microanalysis
  • Fixatives*
  • Formaldehyde
  • Freeze Drying
  • Frozen Sections*
  • Histocytochemistry
  • Histological Techniques*
  • Immunologic Techniques
  • Male
  • Microtomy*
  • Osmium Tetroxide
  • Pancreas / enzymology
  • Pancreas / ultrastructure*
  • Rats
  • Rats, Inbred Lew
  • Volatilization
  • alpha-Amylases / analysis*

Substances

  • Fixatives
  • Formaldehyde
  • alpha-Amylases
  • Osmium Tetroxide