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Harmonemia: a universal strategy for flow cytometry immunophenotyping—A European LeukemiaNet WP10 study

Leukemia volume 30, pages 1769–1772 (2016)Cite this article

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Flow cytometry (FC), a keystone in the diagnosis of hematological malignancies, usefully complements morphology and enables early decisions for patient management.1, 2 Therefore, it is important that FC can be performed in many different centers, in the vicinity of patients. Progress achieved in the past few decades in the stability and precision of flow cytometers and analysis software permit the acquisition of reliable data in any competent laboratory. Owing to the sophistication and versatility of instruments now allowing for multi-color analyzes, and to the large number of reagents available, it remains necessary to define the most pertinent strategies to achieve comparable results between laboratories. A large amount of information is available in the literature concerning the choice of antigens to be tested in hematological diseases.1, 2, 3, 4 Recently, the Euroflow consortium published an extensive description of standardization in the FC diagnosis of hematological neoplasms.5 However, application of these recommendations requires trained personnel for the maintenance of one type of instruments and a specific type of software for data analysis. The proposed panels, extensively tested and calibrated, besides being relatively expensive, are fixed and not likely to be changed if comparison with the pathology library proposed by the consortium is to be applied.

The Harmonemia project described in the current report was set up with the different aim of providing a simple, rapid and robust way to obtain comparable data on various types of FC instruments. It was set to verify that harmonization can be obtained between different instruments in different settings. Information indicating that instruments of the same make or even from different manufacturers can be cross-calibrated is available in the literature.6, 7, 8 In the Harmonemia project, this concept was extended to an international series of platforms. This manuscript reports on the first two steps of this project, namely the harmonization of photomultiplier (PMT) settings and the staining of normal peripheral blood (PB) with a common antibody panel.

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Acknowledgements

This work was supported by Beckman Coulter, which provided all reagents. The authors are especially grateful to Antoine Pacheco, Sue Ann Molero, Jeannine Holden and Markus Kaymer for fruitful discussions.

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Authors and Affiliations

  1. Hematology Laboratory, Bordeaux University Hospital, Bordeaux, France

    F Lacombe & K Allou

  2. Douglass Hanly Moir Pathology, Sonic Healthcare, Sydney, New South Wales, Australia

    E Bernal

  3. Department of Haematology, Addenbrookes Hospital, Cambridge, UK

    D Bloxham

  4. Immunophenotyping Laboratory, Cardiff University, Cardiff, UK

    S Couzens

  5. Department of Hematology Oncology, University of Pavia and IRCCS Policlinico San Matteo, Pavia, Italy

    M G D Porta

  6. Flow Cytometry Laboratory, Bristol Royal Infirmary, Bristol, UK

    U Johansson

  7. München Leukemia Labor, Munich, Germany

    W Kern

  8. St Bartholomew's and Royal London School of Medicine and Dentistry, London, UK

    M Macey

  9. Hematology Laboratory, Geneva University Hospital, Geneva, Switzerland

    T Matthes

  10. The Institute of Cancer Research, Royal Cancer Hospital, London, UK

    R Morilla

  11. Flow Cytometry Unit, Clinical Pathology Service, Coimbra University Hospital Centre, Coimbra, Portugal

    A Paiva

  12. Department of Hematology, Hospital Universitario Vall d’Hebron, Barcelona, Spain

    C Palacio

  13. Department of Laboratory Medicine-Laboratory for Hematology, Radboud University Medical Center, Nijmegen, The Netherlands

    F Preijers

  14. Department of Hematology and Oncology, Helios Klinikum Berlin Buch, Berlin, Germany

    R Ratei

  15. Laboratory Services (HUSLAB), University Central Hospital, University of Helsinki, Helsinki, Finland

    S Siitonen

  16. Department of Pathobiology and Laboratory Medicine, University of Toronto, University Health Network, Toronto General Hospital, Toronto, Ontario, Canada

    A Porwit

  17. Hematology Laboratory, Nantes University Hospital, Nantes, France

    M C Béné

Authors
  1. F Lacombe
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  10. R Morilla
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  17. A Porwit
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  18. M C Béné
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Correspondence to M C Béné.

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The authors declare no conflict of interest.

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Lacombe, F., Bernal, E., Bloxham, D. et al. Harmonemia: a universal strategy for flow cytometry immunophenotyping—A European LeukemiaNet WP10 study. Leukemia 30, 1769–1772 (2016). https://doi.org/10.1038/leu.2016.44

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