Application of the Paired Label Radioantibody Technique to Tissue Sections and Cell Smears

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Abstract
Summary: A micro-scale modification of the paired label radioantibody technique has been developed to determine the reactivity of antiserum globulin with tissue sections or cell smears. A mixture of 125I-labeled IgG from antiserum and 131I-labeled IgG from normal serum is applied on test specimens fixed on microscope coverslips. After incubation for 60 min at room temperature, the specimens are washed and assayed for the radioactivity due to 125I and 131I. The degree of the reactivity of antiserum IgG was expressed by the uptake quotient, the quotient of the specific uptake over the nonspecific uptake. The method is sensitive and reproducible under the conditions given. It is suitable for comparing a large number of specimens and particularly useful where the test materials, tissues or cells, are available only in limited quantities. Various factors involved in the reaction—reaction time, concentration of antibody globulin, area and thickness of sections, antigen content and absorption with tissue sediments—were evaluated in detail.