Multivariate analysis and list mode processing of murine hemopoietic subpopulations for cytokinetic studies

M. G. Pallavicini, L. J. Summers, F. J. Giroud, P. N. Dean, I. W. Gray

Research output: Contribution to journalArticle

Abstract

Multivariate analyses and list mode data processing were used to obtain cytokinetic information on flow cytometrically distinct hemopoietic subpopulations. In one application, viable, unfixed hemopoietic subpopulations were discriminated on the basis of cyanine dye fluorescence and orthogonal light scatter; Hoechst dye fluorescence was measured to determine the proliferative status of the subpopulations. In another application, ethanol‐fixed mouse bone marrow cells were triply stained with Hoechst dye, rhodamine‐conjugated wheat germ agglutinin (WGA), and a fluorescein‐labeled monoclonal antibody against bromodeoxyuridine. In both of these studies, flow cytometric data for all three variables were acquired in list mode fashion, stored on magnetic tape, and processed by list mode software on a computer based multivariable pulse‐height analyzer. In the first application, subpopulations distinguished by cyanine dye intensity and light scatter appeared to be more related to cell lineage and cell size than proliferative status. In the second application, WGA affinity discriminated two subpopulations in mouse bone marrow S‐phase cells in each subpopulation that actively incorporated bromodeoxyuridine (BrdUrd). List mode data processing obviates the need for routine electronic sorting of cells and thus facilitates characterization of discriminated subpopulations. In this regard, it is particularly useful for the study of flow cytometrically distinct, low frequency subpopulations.

Original languageEnglish (US)
Pages (from-to)539-549
Number of pages11
JournalCytometry
Volume6
Issue number6
DOIs
StatePublished - Nov 1985

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Keywords

  • anti‐bromodeoxyuridine antibody
  • cyanine
  • hemopoietic subpopulations
  • Hoechst
  • Multivariate analyses

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Biophysics
  • Hematology
  • Endocrinology
  • Cell Biology

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