The application of a biotin-anti-biotin gold technique providing a significant signal intensification in electron microscopic immunocytochemistry: a comparison with the ultrasmall immunogold silver staining procedure.
A three-step biotin-anti-biotin gold-detection system (method A) has been applied for ultraimmunocytochemistry using ultrasmall colloidal gold (0.8 nm) linked to anti-biotin antibodies which were visualized and enhanced by silver reduction. The reactivity for glucagon in human pancreatic islets and for cytochrome-c oxidase in heart mitochondria has been compared to a two-step ultrasmall immunogold technique (method B). For both antigens, method A provided significantly higher labelling indices (P<0.001): the labelling density for cytochrome-c oxidase was 223/microm2 using method A and 78/microm2 using method B. For glucagon, the labelling density was 1455/microm2 with method A and 322/microm2 with method B. The results demonstrate that the silver-intensified biotin-anti-biotin gold-detection system is a valuable immunocytochemical method for signal enhancement. The method utilizes biotinylated antibodies from different species, allowing its broad application at the electron microscopic level.
Müller-Höcker
J
J
Ludwig-Maximilians University Munich, Germany.
Schäfer
S
S
Sendelhofert
A
A
Weis
S
S
D000818:Animals; D001710:Biotin; D003576:Electron Transport Complex IV / Q000378:metabolism; D005069:Evaluation Studies as Topic; D005934:Glucagon / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D007515:Islets of Langerhans / Q000378:metabolism / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron / Q000379:methods*; D008929:Mitochondria, Heart / Q000201:enzymology / Q000648:ultrastructure; D009206:Myocardium / Q000201:enzymology / Q000648:ultrastructure; D051381:Rats; D012834:Silver; D013194:Staining and Labeling / Q000379:methods
D012834:Silver; D001710:Biotin; D005934:Glucagon; D003576:Electron Transport Complex IV