TY - JOUR
T1 - Multivariate analysis and list mode processing of murine hemopoietic subpopulations for cytokinetic studies
AU - Pallavicini, M. G.
AU - Summers, L. J.
AU - Giroud, F. J.
AU - Dean, P. N.
AU - Gray, I. W.
N1 - Copyright:
Copyright 2016 Elsevier B.V., All rights reserved.
PY - 1985/11
Y1 - 1985/11
N2 - 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.
AB - 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.
KW - Hoechst
KW - Multivariate analyses
KW - anti‐bromodeoxyuridine antibody
KW - cyanine
KW - hemopoietic subpopulations
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U2 - 10.1002/cyto.990060608
DO - 10.1002/cyto.990060608
M3 - Article
C2 - 3840733
AN - SCOPUS:0022185453
VL - 6
SP - 539
EP - 549
JO - Communications in Clinical Cytometry
JF - Communications in Clinical Cytometry
SN - 0196-4763
IS - 6
ER -