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

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 immunoreactivty of α-amylase in exocrine pancreas, as was demonstrated with an indirect labeling technique using anti-α-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.