Immunocytochemistry controls using cell culture

Abstract

A continuing problem in cytology laboratories is the lack of adequate control material for immunocytochemical testing. Usual control procedures involve testing paraffin-embedded control materials along with the patient specimens. These control materials are fundamentally unlike cytologic preparations. We have developed a method to make control preparations for immunocytochemical analysis using cultured anaplastic cells with known antigenic features from commercial sources. Cell lines included melanoma, rhabdomyosarcoma, T-cell leukemia, and squamous-cell carcinoma. Modified Saccomano and acetone fixation coupled with the cytospin technique enabled good-quality preparations. Cell lines were tested with antibodies for HMB-45, actin, leukocyte common antigen (LCA) and cytokeratin, which avidin-biotin immunoperoxidase and diaminobenzidine (DAB) as chromogens. Our final preparations were easily interpretable with excellent morphologic preservation of cellular detail. Cultured cells provide a superior method for preparing almost unlimited numbers of control slides for immunocytochemistry for laboratories with access to a tissue culture facility.