PMID: 2504993Sep 1, 1989Paper

Determination of antigen concentration in tissue sections by immunodensitometry

Laboratory Investigation; a Journal of Technical Methods and Pathology
J RahierJ C Henquin

Abstract

Our aim was to determine whether density of immunolabeling can be used to estimate the amount of an antigen in a tissue. The biological model was the pancreatic insulin-containing B cell. The insulin content of the pancreas of Wistar rats was decreased by five injections of glibenclamide (0.5, 1, or 2 mg/kg) every 12 hours. After resection of the whole pancreas specimens were taken for insulin extraction and measurement by radioimmunoassay and for immunocytochemistry. The sections were treated either by a polyclonal anti-insulin serum at 1/500 or 1/3000 and peroxidase-antiperoxidase complex or by a monoclonal anti-insulin serum at 1/500 and indirect immunoperoxidase. Peroxidase was revealed by diaminobenzidine. The density of immunostained B cells was determined with an automatic image analyzer (Ibas 2000, Kontron, FRG). Compared with controls, pancreatic insulin concentration was decreased by about 40, 60, and 85% in rats treated by the three doses of glibenclamide. A strong correlation was found between the insulin concentration and the optical density of islets under certain conditions: with the monoclonal anti-insulin serum (r = 0.90) and with the polyclonal anti-insulin serum at a high dilution (r = 0.95) but not at a low ...Continue Reading

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