Rapid low molecular weight polyethylene glycol embedding protocol for immunocytochemistry.
We describe an alternative polyethylene glycol (PEG) embedding procedure which utilizes PEG 200 for dehydration and PEG 600 for infiltration and embedding of perfusion-fixed rat liver. PEG 600 has a melting point of 22 degrees C, enabling infiltration of fixed tissue to be performed at room temperature. Sections (2 microM) cut in a cryostat at -20 degrees C and immobilized in agarose were readily labeled by immunoperoxidase protocols with monoclonal antibodies to hepatocyte membrane antigens. Subsequent examination by light microscopy or by electron microscopy after re-embedding in resin and ultra-thin sectioning showed excellent preservation of morphology, with minimal impairment of antigenicity.
Mowery
J
J
Brown University-Rhode Island Hospital, Providence 02902.
Chesner
J
J
Spangenberger
S
S
Hixson
D C
DC
D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis*; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry / Q000379:methods*; D008099:Liver / Q000276:immunology* / Q000648:ultrastructure; D008297:Male; D008854:Microscopy, Electron; D008970:Molecular Weight; D011092:Polyethylene Glycols; D051381:Rats
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D011092:Polyethylene Glycols