Colloidal gold immunostaining on deplasticized ultra-thin sections.
We localized tissue antigens on ultra-thin sections by deplasticizing the sections while on the grid, incubating in primary antiserum followed by immunoglobulin-conjugated colloidal gold, and ultimately re-embedding in dilute Epon. This procedure permitted ultrastructural localization of tissue antigens that were previously masked by the embedding plastic surrounding tissue components. In addition, replacement of the plastic matrix on the thin section after immunostaining prevented development of the drying artifacts that occur in unsupported tissue sections. Optimal preservation of components in the tissue sections was achieved despite extensive steps involved in plastic removal and immunostaining. This method may be useful in situations where the number of exposed epitopes on the surface of a thin section is low. The procedure also allows the use of antisera at greater dilutions and provides enhanced immunostaining specificity with low background.
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Department of Pathology, University of Washington, Seattle 98195.
Wight
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D000199:Actins / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D001158:Arteries / Q000032:analysis / Q000648:ultrastructure; D003102:Colloids; D003594:Cytoplasmic Granules / Q000032:analysis; D006046:Gold; D013006:Growth Hormone / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D007668:Kidney / Q000201:enzymology / Q000648:ultrastructure; D008854:Microscopy, Electron; D009131:Muscle, Smooth, Vascular / Q000032:analysis / Q000648:ultrastructure; D010902:Pituitary Gland / Q000032:analysis / Q000648:ultrastructure; D011509:Proteoglycans / Q000032:analysis; D051381:Rats; D012116:Resins, Plant; D000254:Sodium-Potassium-Exchanging ATPase / Q000032:analysis
D000199:Actins; D000911:Antibodies, Monoclonal; D003102:Colloids; D011509:Proteoglycans; D012116:Resins, Plant; D006046:Gold; D013006:Growth Hormone; D000254:Sodium-Potassium-Exchanging ATPase