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Licensed Unlicensed Requires Authentication Published by De Gruyter July 11, 2019

Preliminary evaluation of a new flow cytometry method for the routine hematology workflow

  • Michela Seghezzi , Valentina Moioli , Giulia Previtali , Barbara Manenti , Ramon-Simon Lopez , Mari Kono , Ezio Tirloni , Maria Grazia Alessio and Sabrina Buoro EMAIL logo



In a generalist laboratory, the integration of the data obtained from hematology analyzers (HAs) with those from multiparametric flow cytometry (FMC) could increase the specificity and sensitivity of first level screening to identify the pathological samples. The aim of this study was to perform a preliminary evaluation of a new simple hybrid method (HM). The method was obtained by integration between HAs reagents into FCM, with a basic monoclonal antibodies panel for the leukocytes differential count.


Eighty-one peripheral blood samples, collected in K3EDTA tubes, were analyzed by XN-module, and CyFlow Space System, using both standard MoAbs and HM method analysis, and with the optical microscopy (OM). Within-run imprecision was carried out using normal samples, the carryover was evaluated, data comparison was performed with Passing-Bablok regression and Bland-Altman plots.


The within-run imprecision of HM methods ranged between 1.4% for neutrophils (NE) and 10.1% for monocytes (MO) always equal or lower to the OM. The comparison between HM methods vs. OM shows Passing-Bablok regression slopes comprised between 0.83 for lymphocyte (LY) and 1.14 for MO, whilst the intercepts ranged between −0.18 for NE and 0.25 for LY. Bland-Altman relative bias was comprised between −12.43% for NE, and 19.77% for eosinophils. In all 11 pathological samples the agreement between the methods was 100%.


The new hybrid method generates a leukocytes differential count suitable for routine clinical use and it is also useful for identifying morphological abnormalities with a reduction in cost and improvement of screening for first level hematology workflow.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: The study was financially supported by Sysmex Corporation and Sysmex America Inc.

  3. Employment or leadership: Dr. Ramon Simon-Lopez is the Consultant Medical Director of Sysmex Corporation and Sysmex America Inc. Dr. Mari Kono is Researcher of Scientific Affairs of Sysmex Corporation and Ezio Tirloni is a Specialist of Product and Application of Sysmex Partec Italia.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.


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Received: 2018-12-20
Accepted: 2019-03-10
Published Online: 2019-07-11
Published in Print: 2019-09-25

© 2019 Walter de Gruyter GmbH, Berlin/Boston

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