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Simultaneous Analysis of DNA and Cell Surface Immunoglobulin in Human B-Cell Lymphomas by Flow Cytometry¹

Department of Pathology, University of Florida College of Medicine, Gainesville, Florida 32610

Cell suspensions prepared from human B-cell lymphomas were analyzed for cellular DNA and cell surface immunoglobulin (Slg) by flow cytometry. In addition to Slg-bearing neoplastic B-lymphocytes, these tumors contain nonneoplastic cells. Flow cytometry permitted selective analysis of DNA in subpopulations defined by the membrane antigen Slg and also permitted analysis of Slg of subpopulations defined by DNA content. Because measurements of DNA and Slg were made simultaneously on each cell, physical separation of cells was not required. Using propidium iodide as a fluorescent DNA probe and fluorescein isothiocyanate-labeled antibodies to stain Slg, differences in cellular DNA between Slg-bearing neoplastic cells and nonneoplastic cells from the same tissues could be demonstrated. Additionally, this simultaneous correlated analysis allowed assessment of the cell cycle dependence of Slg expression. Mean fluorescein isothiocyanate (Slg) fluorescence was greater in S- than G₁-phase cells, but the ratio of mean fluorescein isothiocyanate fluorescence of cells in S phase to that of cells in G₁ phase varied for different tumors.

¹ Supported by Grant CA-23393 from the National Cancer Institute.

² Present address: National Heart, Lung, and Blood Institute, NIH, Bethesda, Md. 20205.

³ To whom requests for reprints should be addressed.

Received 3/27/81. Accepted 8/21/81.