3 PARAMETER FLOW CYTOMETRIC ANALYSIS FOR SIMULTANEOUS DETECTION OF CYTOKERATIN, PROLIFERATION-ASSOCIATED ANTIGENS AND DNA CONTENT

The analysis of the cell cycle distributions by univariate now cytomet ric DNA measurement has been widely applied in the clinic to determine kinetic parameters of human malignancies, A common problem with measu rements of cell cycle phase distributions in tumor biopsy material is the presence of non-malignant diploid cells, Furthermore, such a stati c measurement might not be accurate enough to describe the dynamic pro cess of cell proliferation, For this purpose alternative methods have been developed to include BrdUrd incorporation or the presence of intr insic proliferation associated markers such as PCNA or Ki67-Ag into th e analysis, However, the presence of nonmalignant diploid cells will i nfluence also these bivariate analyses, especially in case of DNA-dipl oidy of the tumor cells. Here we present a three parameter how cytomet ric assay based on the simultaneous detection of cytokeratin, DNA and a proliferation associated marker, such as BrdUrd, PCNA or Ki67-Ag, Ba sed on the presence of cytokeratin, epithelial cells can be selected f or a detailed cell cycle analysis, This method can be applied to froze n tissue, which makes this assay useful for multicentre clinical studi es. (C) 1995 Wiley-Liss, Inc.