Rapid microwave fixation of rat mast cells. I. Localization of granule chymase with an ultrastructural postembedding immunogold technique.
We defined the ultrastructural localization of chymase in rat peritoneal mast cells using standard aldehyde fixation and a newly described microwave fixation method (Login GR, Dvorak AM: Microwave energy fixation for electron microscopy. Am J Pathol 120: 230, 1985; Login GR, Stavinoha WB, Dvorak AM: Ultrafast microwave energy fixation for electron microscopy. J Histochem Cytochem 34:381, 1986) and postembedding immunogold labeling. Thin sections were exposed first to goat IgG anti-rat chymase and second to gold-conjugated rabbit Ig directed against goat IgG. By transmission electron microscopy, gold particles were localized to the matrix of cytoplasmic granules. Control sections treated with nonimmune sera did not exhibit labeling of mast cells. Thin sections treated simultaneously with purified rat mast cell chymase and anti-chymase antibody in competition studies, showed a marked reduction in granule staining. These findings demonstrate that a microwave fixation method can be used to rapidly fix cell suspensions for postembedding immunocytochemical studies.
D000818:Animals; D053818:Chymases; D003594:Cytoplasmic Granules / Q000201:enzymology; D007150:Immunohistochemistry / Q000379:methods*; D008297:Male; D008407:Mast Cells / Q000201:enzymology* / Q000648:ultrastructure; D008854:Microscopy, Electron; D008872:Microwaves; D051381:Rats; D011919:Rats, Inbred Strains; D012697:Serine Endopeptidases / Q000032:analysis*
D012697:Serine Endopeptidases; D053818:Chymases