Vapor fixation for immunocytochemistry and X-ray microanalysis on cryoultramicrotome sections.
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.
D000818:Animals; D004577:Electron Probe Microanalysis; D005404:Fixatives; D005557:Formaldehyde; D005612:Freeze Drying; D005629:Frozen Sections; D006651:Histocytochemistry; D006652:Histological Techniques; D007158:Immunologic Techniques; D008297:Male; D008867:Microtomy; D009993:Osmium Tetroxide; D010179:Pancreas / Q000201:enzymology / Q000648:ultrastructure*; D051381:Rats; D011917:Rats, Inbred Lew; D014835:Volatilization; D000516:alpha-Amylases / Q000032:analysis*
D005404:Fixatives; D005557:Formaldehyde; D000516:alpha-Amylases; D009993:Osmium Tetroxide