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The optimal application of forward and ninety-degree light scatter in flow cytometry for the gating of mononuclear cells†
Article first published online: 8 MAR 2005
Copyright © 1985 Wiley-Liss, Inc.
Volume 6, Issue 5, pages 401–406, September 1985
How to Cite
Thompson, J. M., Gralow, J. R., Levy, R. and Miller, R. A. (1985), The optimal application of forward and ninety-degree light scatter in flow cytometry for the gating of mononuclear cells. Cytometry, 6: 401–406. doi: 10.1002/cyto.990060503
- Issue published online: 16 JUN 2005
- Article first published online: 8 MAR 2005
- Manuscript Accepted: 1 APR 1985
- Manuscript Received: 10 JAN 1985
- Light scatter;
- flow cytometry;
- computerized gating
Peripheral blood mononuclear cells from ten normal donors were labeled with a monoclonal antibody specific for monocytes and analyzed using a fluorescence activated cell sorter (FACS). Forward and 90° light scatter parameters were studied in order to apply optimal computerized gating to identify and exclude monocytes from lymphocyte populations. An average of 9.45% versus 1.22% of cells, within chosen lymphocyte gates established by forward angle and 90° scatter, respectively, were identified as monocytes. In samples from ten donors, the exclusion of monocytes from the lymphocyte population was more efficient using 90° scatter than forward scatter. Simultaneous use of forward and 90° scatter did not significantly improve the ability to accurately exclude monocytes, but did result in a significant increase in the improper exclusion of lymphocytes. Use of 90° scatter alone, forward scatter alone, and forward and 90° scatter simultaneously to identify lymphoid cells resulted in the exclusion of 12, 17, and 23% of lymphocytes from further analysis. The 90° scatter alone appears to be the optimal method to eliminate monocytes electronically from mononuclear cell populations in which lymphocytes are being studied.