Three parameter flow cytometric analysis for simultaneous detection of cytokeratin, proliferation associated antigens and DNA content

Journal Article · Sun Oct 01 04:00:00 EDT 1995 · Cytometry

DOI:https://doi.org/10.1002/cyto.990210210· OSTI ID:456816

Schutte, B; Pijpers, G F.P.; Lenders, M H.J.H.; Ramaekers, F C.S. ^[1]

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1. Univ. of Limburg (Netherlands)

The analysis of the cell cycle distributions by univariate flow cytometric DNA measurement has been widely applied in the clinic to determine kinetic parameters of human malignancies. A common problem with measurements of cell cycle phase distributions in tumor biopsy material is the presence of nonmalignant diploid cells. Furthermore, such a static measurement might not be accurate enough to describe the dynamic process of cell proliferation. For this purpose alternative methods have been developed to include BrdUrd incorporation or the presence of intrinsic proliferation associated markers such as PCNA or Ki67-Ag into the analysis. However, the presence of nonmalignant diploid cells will influence also these bivariate analyses, especially in case of DNA-diploidy of the tumor cells. Here we present a three parameter flow cytometric assay based on the simultaneous detection of cytokeratin, DNA and a proliferation associated marker, such as BrdUrd, PCNA or Ki67-Ag. Based on the presence of cytokeratin, epithelial cells can be selected for a detailed cell cycle analysis. This method can be applied to frozen tissue, which makes this assay useful for multicentre clinical studies. 25 refs., 8 figs.

• Sponsoring Organization: USDOE
• OSTI ID: 456816
• Journal Information: Cytometry, Journal Name: Cytometry Journal Issue: 2 Vol. 21; ISSN CYTODQ; ISSN 0196-4763
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
ANTIBODIES
ANTIGENS
BIOASSAY
BIOCHEMICAL REACTION KINETICS
BUDR
CELL CYCLE
CELL FLOW SYSTEMS
CELL PROLIFERATION
DETECTION
DIPLOIDY
DNA
NEOPLASMS
SAMPLE PREPARATION
THERAPY
TUMOR CELLS