Use of poligonal windows for physical discrimination among mononuclear subpopulations in flow cytometry

Marco Vitale, Stefano Papa, Adriana Rita Mariani, Andrea Facchini, Riccardo Rizzoli, Francesco A. Manzoli

Research output: Contribution to journalArticle

Abstract

Peripheral blood mononuclear cells from ten healthy normal donors were analyzed by means of monoclonal antibodies and flow cytometry (FACS IV). In order to apply optimal gating for the identification and exclusion of monocytes from lymphocyte populations, mononuclear cells were analyzed on the scattergram using rectangular or poligonal computer-generated windows. Two main windows on lymphoid population were used which mainly differed for the up-right corner, in the border area between lymphocytes and monocytes. While the number of lymphocytes contained either in the regular or in the poligonal windows was the same, the number of contaminating monocytes decreased by two-fold in the poligonal one. Besides, the use of tight lymphocytes gating, in order to reach lower monocyte contamination, leads to a loss of lymphoid cells which does not appear to be random, but seems to affect mainly the Leu11c+ population with natural killer activity. These cells produce forward and perpendicular scatter signals higher than other lymphocyte subsets, and, therefore, are mainly located in the area of the scattergram which divides lymphocytes from monocytes. These data are in accordance with the large granular morphology of natural killer cells. The use of the poligonal windows seems to be useful to reduce monocyte contamination with no selective loss of natural killer lymphocytes, and may be particularly helpful in the analysis of pathological samples.

Original languageEnglish
Pages (from-to)63-68
Number of pages6
JournalJournal of Immunological Methods
Volume96
Issue number1
DOIs
Publication statusPublished - Jan 26 1987

Keywords

  • Cell sorter
  • fluorescence-activated
  • Immunofluorescence
  • NK cell
  • Scatter signal

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

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