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Keywords:

  • B-cell chronic lymphoproliferative disorders;
  • flow cytometry;
  • immunophenotyping;
  • FCS files;
  • data calculation;
  • nearest neighbor

Abstract

Immunophenotypic characterization of B-cell chronic lymphoproliferative disorders (B-CLPD) is associated with the use of increasingly larger panels of multiple combinations of 3 to ≥6 monoclonal antibodies (Mab), data analysis being separately performed for each of the different stained sample aliquots. Here, we describe and validate an automated method for calculation of flow cytometric data from several multicolor stainings of the same cell sample—i.e., the merging of data from different aliquots stained with partially overlapping combinations of Mab reagents (focusing on ≥1 cell populations)—into one data file as if it concerned a single “super” multicolor staining. Evaluation of the performance of the method described was done in a group of 60 B-CLPD studied at diagnosis with 18 different reagents in a panel containing six different 3- and 4-color stainings, which systematically contained CD19 for the identification of B-cells. Our results show a high degree of correlation and agreement between originally measured and calculated data about cell surface stainings, providing a basis for the use of this approach for the generation of flow cytometric data files containing information about a virtually infinite number of stainings for each individual cellular event measured in a sample, using a limited number of fluorochrome stainings. © 2008 International Society for Advancement of Cytometry